Search

Parkinson’s disease (PD) is characterized by the loss of dopaminergic neurons in the substantia nigra, causing motor changes. In addition to motor symptoms, non-motor dysfunctions such as psychological, sensory and autonomic disorders are recorded. Manifestations related to the autonomic nervous system include the cardiovascular system, as postural hypotension, postprandial hypotension, and low blood pressure. One of the mediators involved is the nitric oxide (NO). In addition to the known roles such as vasodilator, neuromodulator, NO acts as an important mediator of the immune response, increasing the inflammatory response provoked by PD in central nervous system. The use of non-specific NOS inhibitors attenuated the neurodegenerative response in animal models of PD. However, the mechanisms by which NO contributes to neurodegeneration are still not well understood. The literature suggest that the contribution of NO occurs through its interaction with superoxides, products of oxidative stress, and blocking of the mitochondrial respiratory chain, resulting in neuronal death. Most studies involving Parkinsonism models have evaluated brain NO concentrations, with little data available on its peripheral action. Considering that studies that evaluated the involvement of NO in the neurodegeneration in PD, through NOS inhibitors administration, showed neuroprotection in rats, it has prompted new studies to assess the participation of NOS isoforms in cardiovascular changes induced by parkinsonism, and thus to envision new targets for the treatment of cardiovascular disorders in PD. The aim of this study was to conduct a literature review to assess available information on the involvement of nitric oxide (NO) in cardiovascular aspects of PD.

We sought to investigate whether RVLM iNOS activity and oxidative profile may participate in the reduction of sympathetic responsiveness in swimming trained normotensive rats. Sedentary (S) and swimming trained (T) Wistar male rats chronically instrumented with an arterial catheter and guide cannula into the RVLM were submitted to continuous pressure and heart rate (HR) recordings and determination of autonomic control (power spectral analysis) before and after unilateral RVLM iNOS inhibition (aminoguanidine, 250 pmol/100 nL). Other S and T rats received local l-glutamate microinjection (5 nmol/100 nL). In separate S and T groups not submitted to brainstem cannulation, fresh bilateral RVLM punchs were collected for iNOS gene expression (qPCR); reduced glutathione and lipid peroxidation quantification (spectrophotometry); iron-reducing antioxidant (FRAP) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical cation (ABTS˙+) scavenger assays. iNOS gene expression was confirmed in fixed RVLM slices (immunofluorescence). T rats exhibited resting bradycardia, lower sympathovagal balance, reduced RVLM iNOS gene/protein expression and higher antioxidant capacity. Decreased iNOS expression was positively correlated with reduced HR. Pressor and tachycardic response to l-Glutamate were smaller in T rats. Aminoguanidine microinjection reduced sympathetic activity in S rats but did not change it in T rats expressing reduced RVLM iNOS content. Our data indicate that iNOS, expressed in the RVLM of normotensive male rats, has tonic effects on sympathetic activity and that swimming training is an efficient tool to reduce iNOS expression and augment the antioxidant defense, thus reducing glutamatergic responsiveness and sympathetic drive to cardiovascular effectors.

Chagas disease, caused by the protozoan Trypanosoma cruzi, is one of the main causes of death due to cardiomyopathy and heart failure in Latin American countries. The treatment of Chagas disease is directed at eliminating the parasite, decreasing the probability of cardiomyopathy and disrupting the disease transmission cycle. Benznidazole (BZ) and nifurtimox (Nfx) are recognized as effective drugs for the treatment of Chagas disease by the World Health Organization, but both have high toxicity and limited efficacy, especially in the chronic disease phase. At low doses, aspirin (ASA) has been reported to protect against T. cruzi infection. We evaluated the effectiveness of BZ in combination with ASA at low doses during the acute disease phase and evaluated cardiovascular aspects and cardiac lesions in the chronic phase. ASA treatment prevented the cardiovascular dysfunction (hypertension and tachycardia) and typical cardiac lesions. Moreover, BZ+ASA-treated mice had a smaller cardiac fibrotic area than BZ-treated mice. These results were associated with an increase in numbers of eosinophils and reticulocytes and levels of nitric oxide in the plasma and cardiac tissue of ASA-treated mice relative to respective controls. These effects of ASA and BZ+ASA in chronically infected mice were inhibited by pretreatment with the lipoxin A(4) (LXA(4)) receptor antagonist Boc-2, indicating that the protective effects of ASA are mediated by ASA-triggered lipoxin. These results emphasize the importance of exploring new drug combinations for treatments of the acute phase of Chagas disease that are beneficial for patients with chronic disease.

The rostral ventrolateral medulla (RVLM) is the main sympathetic output of the central nervous system to control blood pressure. Reportedly, reactive oxygen species (ROS) can increase arterial pressure, leading to hypertension. As ROS increase the sympathetic tone in RVLM and obese animals present grater oxidative stress, it would be important to note this relationship.Therefore, we evaluated the systemic and central effects (in the RVLM) of vitamin C (vit C, an antioxidant) on the redox balance and cardiovascular and autonomic profiles in hyperadipose male rats. We also evaluated the neurotransmission by L-glutamate (L-glu) and vit C in the RVLM of awake hyperadipose rats.Our study confirmed that hyperadipose rats were hypertensive and tachycardic, presented increased sympathetic and decreased parasympathetic modulation of the heart, and had increased plasma lipoperoxidation compared with the control rats (CTR). Oral vitamin C treatment reverted cardiovascular, autonomic, and plasma redox dysfunction. Hyperadipose rats presented a higher blood pressure increase after L-glu microinjection and a lower response to vit C in the RVLM compared with the CTR group. Biochemical analysis of redox balance in RVLM punches showed that hyperadipose rats have increased NBT and T-BARS, and after treatment with vit C, the oxidative profile decreased. The antioxidative activity of vit C reduced the amount of ROS in the RVLM area that might have resulted in lowered blood pressure and sympathetic modulation.Our data suggest central and peripheral benefits of vit C treatment on cardiovascular, autonomic, and oxidative dysfunctions in hyperadipose animals.

Objective: The aim was to evaluate if maternal treatment with metformin (MET) during pregnancy and lactation could be safe for metabolic and cardiovascular parameters of adult male and female offsp...

Made available in DSpace on 2018-12-11T17:20:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-07-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Studies showed that physical exercise decreases the risk of developing Parkinson's disease (PD) as slowing its progression. Nitric oxide (NO) increases in the substantia nigra pars compacta (SNpc) of individuals with PD. However, no study has evaluated the effects of exercise on peripheral NO levels and its modulatory effects on cardiovascular dysfunctions of subjects with PD. Trained (T) or sedentary (S) animals underwent stereotactic surgery for bilateral 6-hydroxydopamine (6-OHDA) or vehicle microinfusion (Sham group). After 6 days, the animals were catheterized for baseline parameters, followed by inhibition of NOS by Nw-nitro-arginine-methyl ester (L-NAME, 10 mg/kg - i.v.). Nitrite concentration was performed in the aorta, heart, kidney, adrenal and plasma. After exercise, the animals presented resting bradycardia (6-OHDA T and Sham T). NO was increased in the aorta of 6-OHDA S, and decreased in 6-OHDA T animals. In the heart, NO was increased in Sham T compared to sedentary and decreased in 6-OHDA T relative to 6-OHDA S and Sham T animals. At the kidney, NO decrease in 6-OHDA S and Sham T when compared to Sham S and, in adrenal gland, there was a decrease in 6-OHDA T in relation to 6-OHDA S. L-NAME promoted lower increases in MAP in 6-OHDA groups. The decreases of HR were enhanced due to physical training. 6-OHDA S group presented decreased systolic arterial pressure variability, not altered by exercise. Our data showed alterations in peripheral NO in the association of exercise with Parkinsonism in the cardiovascular function. Departamento de Ciências Fisiológicas Universidade Estadual de Londrina – UEL Faculdade de Ciências Farmacêuticas de Araraquara Departamento de Princípios Ativos Naturais e Toxicologia Universidade Estadual Paulista – UNESP Departamento de Ciências Patológicas Universidade Estadual de Londrina – UEL Faculdade de Ciências Farmacêuticas de Araraquara Departamento de Princípios Ativos Naturais e Toxicologia Universidade Estadual Paulista – UNESP CAPES: 1583617

Aims Sepsis is a potentially fatal systemic inflammatory response and its underlying pathophysiology is still poorly understood. Studies suggest that obesity, a component of metabolic syndrome (MS), is associated with sepsis survival. Therefore, this study focused on investigating the influence of MS on mortality and cardiovascular dysfunction induced by sublethal cecal ligation and puncture (SL-CLP). Main methods Newborn Swiss mice received monosodium glutamate (MSG) (4 mg kg−1 day−1, s.c.) during the first 5 d of life for MS induction, while the control pups received equimolar saline solution. On the 75th day, SL-CLP was used to induce mild sepsis (M-CLP) in the MS (MS-M-CLP) and control (SAL-M-CLP) mice. The effect of MS on sepsis in mice was assessed by determining the survival rate and quantification of nitric oxide (NO) in the plasma, and associating this data with hematological and cardiovascular parameters. Key findings MS improved the survival of septic mice, preventing impairment to hematological and cardiovascular parameters. In addition, MS attenuated plasmatic NO increase, which is a typical feature of sepsis. Significance These findings provide new insights into the relationship between obesity and mild sepsis in mice, thus revealing an approach in favor of the “obesity paradox.”

It is known that autonomic modulation is responsive to ovarian hormone levels and that estrogen increases nitric oxide (NO) bioavailability. However, little is known about the interaction of nitric oxide synthase (NOS) isoforms with autonomic modulation, oxidative stress and cardiovascular risk in females. This study aimed to investigate cardiovascular, autonomic and oxidative parameters after selective NOS inhibition. A spectral analysis of systolic arterial pressure (SAP) and heart rate variability (HRV) was performed. NO levels, total antioxidant capacity (TRAP), lipid hydroperoxides (LOOH) and paraoxonase 1 (PON1) activity were measured in the plasma of rats treated with L-NG-nitroarginine methyl ester (L-NAME), S-methylisothiourea (SMT) or saline. Wistar rats, ovariectomized (OVX) with or without estradiol treatment (1mg/kg/day) or with a false ovariectomy (SHAM), were submitted to artery and vein catheterization. Cardiovascular parameters were evaluated before and after the administration of saline or NOS inhibitors. After 2h, plasma samples were collected for biochemical measurement. At baseline, cardiovascular and autonomic parameters were not different among the groups. L-NAME, the constitutive NOS isoform (cNOS) inhibitor, promoted an increase in mean arterial pressure (MAP) and a reduction in the low frequency band (LF) of SAP of SHAM rats, but this increase was smaller in OVX animals, which also showed a reduction in PON1 activity. The decreased activity of PON1 caused by L-NAME was prevented in the OVX+E group. SMT, an inducible NOS isoform (iNOS) inhibitor, promoted an increase in MAP and in the LF of SAP, in interbeat interval (IBI) parameters at LFnu and in LF/HF ratio of HRV in all groups, but the OVX+E had lower levels of NO when compared with the OVX group. Our data suggest that while cNOS contributes to maintaining the activity of PON1 in OVX rats, iNOS activity maintains the levels of NO in OVX+E rats.

Long-chain omega-3 polyunsaturated fatty acids (LC n-3 PUFA) are known to modulate a variety of immune cell functions. On occasion, this has led to diminished host resistance to certain viral and bacterial infections. Little is known about the impact of n-3 PUFA on host resistance to parasitic infection, however, based on results from a small study conducted more than two decades ago, we hypothesized that providing mice LC n-3 PUFA will diminish host resistance to Trypanosoma cruzi, the parasitic pathogen responsible for Chagas disease. To investigate this, C57BL/6 mice were supplemented by gavage (0.6% v/w) with phosphate-buffered saline, corn oil (CO), or menhaden fish oil (FO, a fat source rich in LC n-3 PUFA) for 15 days prior to T cruzi (Y strain) challenge and throughout the acute phase of infection. FO supplementation was associated with a transient 2-fold greater peak of blood parasitemia at 7 days postinfection (dpi), whereas subsequent cardiac parasitemia was ~60% lower at 12 dpi. FO treatment also ameliorated the leukopenia and thrombocytopenia observed in the early stages of a T cruzi infection. FO supplementation reduced circulating and cardiac nitric oxide at 7 and 12 dpi, respectively. FO supplementation altered ex vivo prostaglandin E2 and cytokine and chemokine production by splenocytes isolated from uninfected and infected mice. Overall, our results suggest that oral administration of LC n-3 PUFA from FO can have beneficial effects on the host in the early course of a T cruzi infection.

We evaluated the influence of metabolic syndrome (MS) on acute Trypanosoma cruzi infection. Obese Swiss mice, 70 days of age, were subjected to intraperitoneal infection with 5 × 102 trypomastigotes of the Y strain. Cardiovascular, oxidative, inflammatory, and metabolic parameters were evaluated in infected and non-infected mice. We observed higher parasitaemia in the infected obese group (IOG) than in the infected control group (ICG) 13 and 15 days post-infection. All IOG animals died by 19 days post-infection (dpi), whereas 87.5% of the ICG survived to 30 days. Increased plasma nitrite levels in adipose tissue and the aorta were observed in the IOG. Higher INF-γ and MCP-1 concentrations and lower IL-10 concentrations were observed in the IOG compared to those in the ICG. Decreased insulin sensitivity was observed in obese animals, which was accentuated after infection. Higher parasitic loads were found in adipose and hepatic tissue, and increases in oxidative stress in cardiac, hepatic, and adipose tissues were characteristics of the IOG group. Thus, MS exacerbates experimental Chagas disease, resulting in greater damage and decreased survival in infected animals, and might be a warning sign that MS can influence other pathologies.

Made available in DSpace on 2021-06-25T12:22:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-10-15 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Aims: Parkinson's disease (PD) is a neurological disorder caused by environmental and genetic factors, char-acterized by the death of dopaminergic neurons of the substantia nigra pars compacta (SNpc), leading to a decrease of dopamine in the striatum. In addition to motor symptoms, PD has several abnormalities, among which are cardiovascular changes, such as orthostatic and postprandial hypotension, and blood pressure lability. Studies demonstrate gender differences in PD pathogenesis, indicating that female hormones have a protective role against disease development. However, no studies examining cardiovascular changes in a female rat model of parkinsonism exist. Main methods: Wistar female rats were subjected to ovariectomy (OVX) or sham surgery. After seven days, these animals were subjected to bilateral infusion of 6-hydroxydopamine (6-OHDA) or vehicle solution in their SNpc. On the 14th experimental day, a femoral artery catheterization was performed to record cardiovascular parameters after 24 h in conscious state. Analyses of cardiovascular variability and spontaneous baroreflex were performed. The nitrite (NO) concentration in the heart, thoracic aorta, abdominal aorta, and plasma was measured. Key findings: The sham-6-OHDA group had no decrease in the mean arterial pressure compared to sham-saline group, whereas the OVX-6-OHDA group presented a baseline decrease in comparison to sham-6-OHDA. The OVX-6-OHDA group showed an NO increase in the heart and abdominal aorta, whereas the sham-6-OHDA group did not. The very low frequency variability component decreased in the sham-6-OHDA but not in the OVX-6OHDA group. Significance: We suggest a cardiovascular protection by ovarian hormones in PD with a possible NO involvement. Univ Estadual Londrina, Ctr Biol Sci, Dept Physiol Sci, Rodovia Celso Garcia Cid,Km 380,Campus Univ, BR-86055900 Londrina, Parana, Brazil Univ Estadual Londrina, Ctr Biol Sci, Dept Pathol Sci, Londrina, Parana, Brazil Sao Paulo State Univ, Fac Pharmaceut Sci, Dept Nat Act Principles & Toxicol, Araraquara, SP, Brazil Sao Paulo State Univ, Fac Pharmaceut Sci, Dept Nat Act Principles & Toxicol, Araraquara, SP, Brazil

Infection with the protozoanTrypanosoma cruzicauses Chagas disease and consequently leads to severe inflammatory heart condition; however, the mechanisms driving this inflammatory response have not been completely elucidated. Nitric oxide (NO) is a key mediator of parasite killing inT. cruzi-infected mice, and previous studies have suggested that leukotrienes (LTs) essentially regulate the NO activity in the heart. We used infected 5-lipoxygenase-deficient mice (5-LO−/−) to explore the participation of nitric oxide synthase isoforms, inducible (iNOS) and constitutive (cNOS), in heart injury, cytokine profile, and oxidative stress during the early stage ofT. cruziinfection. Our evidence suggests that the cNOS of the host is involved in the resistance of 5-LO−/−mice duringT. cruziinfection. iNOS inhibition generated a remarkable increase inT. cruziinfection in the blood and heart of mice, whereas cNOS inhibition reduced cardiac parasitism (amastigote nests). Furthermore, this inhibition associates with a higher IFN-γproduction and lower lipid peroxidation status. These data provide a better understanding about the influence of NO-interfering therapies for the inflammatory response towardT. cruziinfection.

Aim: Autonomic modulation responds to ovarian hormones and estrogen increases nitric oxide bioavailability. Also, females have minor susceptibility to sepsis and a higher survival rate. However, few studies have evaluated the role of estrogen in cardiovascular, autonomic, and oxidative parameters during initial endotoxemia and under inducible nitric oxide synthase (iNOS) inhibition in female rats. Methods: Female wistar rats were subjected to ovariectomy and divided into three groups: OVX (ovariectomized), OVX+E (OVX plus daily estradiol) and SHAM (false surgery). After 8 weeks, mean arterial pressure (MAP) and heart rate (HR) were recorded in non-anesthetized catheterized rats, before and after intravenous LPS injection, preceded by S-methylisothiourea sulfate (SMT) injection, or sterile saline. Cardiovascular recordings underwent spectral analysis for evaluation of autonomic modulation. Two hours after LPS, plasma was collected to assess total radical-trapping antioxidant (TRAP), nitrite levels (NO2), lipoperoxidation (LOOH), and paraoxonase 1 (PON1) activity. Results: Two hours after LPS, females treated with SMT presented a decrease of MAP, when compared to saline-LPS groups. At this same time, all SMT+LPS groups presented an increase of sympathetic and a decrease of parasympathetic modulation of HR. Two hours after saline+LPS, OVX presented decreased total radical-trapping antioxidant (TRAP) compared to SHAM. When treated with SMT+LPS, OVX did not altered TRAP, while estradiol reduced LOOH levels. Conclusion: iNOS would be responsible for sympathetic inhibition and consumption of antioxidant reserves of females during endotoxemia, since iNOS is inhibited, treatment with estradiol could be protective in inflammatory challenges.

AIM Parkinson's disease (PD) is a progressive neurodegenerative disease that manifests itself clinically after reaching an advanced pathological stage. Besides motor signals, PD patients present cardiovascular and autonomic alterations. Recent data showed that rats induced to Parkinsonism by 6-hydroxydopamine (6-OHDA) administration in the substantia nigra pars compacta (SNpc) showed lower mean arterial pressure (MAP) and heart rate (HR), as reduction in sympathetic modulation. The paraventricular nucleus of the hypothalamus (PVN) is an important site for autonomic and cardiovascular control, and amino acid neurotransmission has a central role. We evaluate PVN amino acid neurotransmission in cardiovascular and autonomic effects of 6-OHDA Parkinsonism. METHODS Male Wistar rats were submitted to guide cannulas implantation into the PVN. 6-OHDA or sterile saline (sham) was administered bilaterally in the SNpc. After 7 days, cardiovascular recordings in conscious state was performed. RESULTS Bicuculline promoted an increase in MAP and HR in sham group and exacerbated those effects in 6-OHDA group. NBQX (non-NMDA inhibitor) did not promote changes in sham as in 6-OHDA group. On the other hand, PVN microinjection of LY235959 (NMDA inhibitor) in sham group did not induced cardiovascular alterations, but decreased MAP and HR in 6-OHDA group. Compared to Sham group, 6-OHDA lesion increased the number of neuronal nitric oxide synthase (nNOS)-immunoreactive neurons in the PVN and, nNOS inhibition promoted higher increases in MAP and HR. CONCLUSION Our data suggest that the decreased baseline blood pressure and heart rate in animals with Parkinsonism may be due to an increased GABAergic tone via nNOS in the PVN.

Epigenetics has recently been linked to molecular adaptive responses evoked by physical exercise and stress. Herein we evaluated the effects of physical exercise on global DNA methylation and expression of the Dnmt1 gene in the rat brain and also verified its potential to modulate responses evoked by repeated restraint stress (RRS). Wistar rats were classified into the following experimental groups: (1) physically active (EX): animals submitted to swimming during postnatal days 53-78 (PND); (2) stress (ST): animals submitted to RRS during 75-79PND; (3) exercise-stress (EX-ST): animals submitted to swimming during 53-78PND and to RRS during 75-79PND, and (4) control (CTL): animals that were not submitted to intervention. Samples from the hippocampus, cortex and hypothalamus were obtained at 79PND. The global DNA methylation profile was assessed using an ELISA-based method and the expression of Dnmt1 was evaluated by real-time PCR. Significantly increased methylation was observed in the hypothalamus of animals from the EX group in comparison to CTL. Comparative analysis involving the EX-ST and ST groups revealed increased global DNA methylation in the hippocampus, cortex, and hypothalamus of EX-ST, indicating the potential of physical exercise in modulating the responses evoked by RRS. Furthermore, decreased expression of the Dnmt1 gene was observed in the hippocampus and hypothalamus of animals from the EX-ST group. In summary, our data indicate that physical exercise affects DNA methylation of the hypothalamus and might modulate epigenetic responses evoked by RRS in the hippocampus, cortex, and hypothalamus.

Objective Evaluate the effects caused by L-DOPA on cardiovascular and autonomic parameters in an animal model of Parkinsonism induced by 6-hydroxydopamine (6-OHDA). Methods Adult male Wistar rats were subjected to bilateral microinfusion of 6-OHDA or saline (sham group) in the substantia nigra, and treated by gavage with L-DOPA or water for 7 days after surgery. On the 6th day the rats were subjected to femoral artery catheterization for cardiovascular recording. Mean arterial pressure (MAP) and heart rate (HR) were evaluated at baseline and during head up tilt (HUT) protocol. Spectral analysis of cardiovascular variability was performed using the V2.4 CardioSeries software v2.4. The lesion was quantified by dopamine levels in the striatum. Results Dopamine levels in the striatum were decreased in 6-OHDA rats (sham: 4.79 ± 0.49 ng/mg; 6-OHDA: 1.99 ± 0.68 ng/mg) and were not recovered by Prolopa treatment. Baseline values of MAP and HR were not different between groups. HUT induced an increase in MAP and HR (ΔMAP: 17 ± 1 mm Hg, ΔHR: 39 ± 4 bpm) that were attenuated in 6-OHDA and in Prolopa treated animals. At baseline, the systolic arterial pressure (SAP) variance was lower in the 6-OHDA and sham Prolopa groups. Spontaneous baroreflex sensitivity was higher at baseline in the 6-OHDA group as compared to all studied groups. Conclusions Our data suggest that treatment with Prolopa did not interfere with cardiovascular variables at baseline. However, during HUT, the 6-OHDA and Prolopa control animals presented a lower cardiovascular compensation, suggesting a possible autonomic impairment in Parkinsonism induced by 6-OHDA.

Exposure to microgravity induces autonomic and vestibular disorders such as alterations in cardiovascular function. The paraventricular nucleus of the hypothalamus (PVN) is known to be an important center for integrating autonomic and cardiovascular responses as blood volume reflexes. The acute effects promoted by microgravity and PVN involvement in cardiovascular and autonomic parameters have not yet been evaluated. Male Wistar rats were anesthetized to facilitate cannulae implantation in the PVN. After 3 days of surgical recovery, femoral artery and vein catheters were implanted for direct recording of blood pressure and heart rate (HR) in conscious animals to evaluate cardiovascular and autonomic changes in an acute protocol of head-down tilt (HDT) in nonanesthetized rats. During HDT, there was an increase in mean arterial pressure (11 ± 1 mmHg, P

Nitric oxide (NO), an intercellular signaling molecule is relevant for circulatory autonomic control. Brain NO synthase (NOS) and NO levels were downregulated in pathological conditions, but rescued after exercise training. We hypothesized that exercise training was also able to improve NO modulation within the hypothalamic paraventricular nucleus (PVN) of healthy rats. Male Wistar rats were submitted to two 4-weeks protocols: i) swimming training (T) or kept sedentary (S), ii) l-arginine (62,5 mg/mL, 1 mL/day p. o.) or vehicle supplementation. Rats underwent stereotaxic surgery (PVN bilateral guide cannulas) and chronic catheterization of artery/vein. Arterial pressure (AP), heart rate (HR) and baroreflex sensitivity were recorded in conscious rats at rest and following a selective nNOS inhibitor (Nw-Propyl-l-Arginine, 4 nmol/100 nL) within the PVN. Rats were deeply anesthetized for brain perfusion/harvesting after respiratory arrest. In separate groups (T and S, l-arginine and Vehicle supplemented) not submitted to PVN cannulation, fresh and fixed brains were obtained for gene and protein nNOS expression (qPCR and immunohistochemistry) and nitrite levels (Griess reaction). T and l-arginine treatment were accompanied by resting bradycardia, augmented parasympathetic and reduced sympathetic activity to heart and vessels (power spectral analysis) and increased baroreflex sensitivity (†P < 0.05). In contrast, PVN nNOS inhibition blocked/attenuated these effects in addition to significantly increase in resting MAP and HR (with larger effects in T and l-arginine treated rats vs. respective controls, †P < 0.05). T increased nNOS gene and protein expression within the ventromedial and posterior PVN nuclei (†P < 0.05). PVN nitirite levels were also increased in T and l-arginine groups (†P < 0.05). Data strongly suggest that training by increasing NO availability within PVN preautonomic nuclei favors both the slow down of sympathetic and the augmentation of parasympathetic activity and facilitates baroreflex control, therefore improving autonomic regulation of the heart in healthy rats.

It has been suggested that nitric oxide (NO) from iNOS source is involved in inflammation and oxidative stress, and hypertension in obese subjects involves an inflammatory process. However, no study evaluated the participation of iNOS inhibition on cardiovascular, autonomic, and inflammatory parameters in obese rats. Obesity was induced by the administration of 4 mg/g body weight of monosodium glutamate (MSG) or equimolar saline (CTR) in newborn rats. On the 60th day, treatment with aminoguanidine (Amino, 50 mg/kg), an iNOS inhibitor, or 0.9% saline, was started. On the 90th day, mean arterial pressure (MAP) and heart rate (HR) were recorded in conscious rats and autonomic modulation was conducted with the CardioSeries software. Plasma samples were collected to assess lipid peroxidation and prostaglandins (PGE

We evaluated the effects of swimming training on nitric oxide (NO) modulation to glutamate microinjection within the rostral ventrolateral medulla (RVLM) in conscious freely moving rats. Male Wistar rats were submitted to exercise training (Tr) by swimming or kept sedentary (Sed) for 4 weeks. After the last training session, RVLM guide cannulas and arterial/venous catheters were chronically implanted. Arterial pressure (AP), heart rate (HR), and baroreflex control of HR (loading/unloading of baroreceptors) were recorded in conscious rats at rest. Pressor response to L-glutamate in the RVLM was compared before and after blockade of local nitric oxide (NO) production. In other Tr and Sed groups, brain was harvested for gene (qRT-PCR) and protein (immunohistochemistry) expression of NO synthase (NOS) isoforms and measurement of NO content (nitrite assay) within the RVLM. Trained rats exhibited resting bradycardia (average reduction of 9%), increased baroreflex gain (Tr: −4.41 ± 0.5 vs. Sed: −2.42 ± 0.31 b/min/mmHg), and unchanged resting MAP. The pressor response to glutamate was smaller in the Tr group (32 ± 4 vs. 53 ± 2 mmHg, p < 0.05); this difference disappeared after RVLM pretreatment with carboxy-PTIO (NO scavenger), Nw-Propyl-L-Arginine and L-NAME (NOS inhibitors). eNOS immunoreactivity observed mainly in RVLM capillaries was higher in Tr, but eNOS gene expression was reduced. nNOS gene and protein expression was slightly reduced (−29 and −9%, respectively, P > 0.05). Also, RVLM NO levels were significantly reduced in Tr (−63% vs. Sed). After microinjection of a NO-donor, the attenuated pressor response of L-glutamate in Tr group was restored. Data indicate that swimming training by decreasing RVLM NO availability and glutamatergic neurotransmission to locally administered glutamate may contribute to decreased sympathetic activity in trained subjects.

Parkinson's disease (PD) is mainly characterized by motor signals. However, non-motor signals also affect and decrease the quality of life of PD patients. Among these non-motor signs are cardiovascular disorders as orthostatic hypotension, postprandial hypotension and cardiac arrhythmias, which may be due to the involvement of both central nervous system and peripheral autonomic nervous system. In the present study we investigated the cardiovascular function, evaluating cardiovascular reflexes (chemoreflex and baroreflex), in an animal model of Parkinsonism induced by bilateral infusion of the toxin 6-hydroxydopamine (6-OHDA), in the substantia nigra pars compacta (SNpc). The results showed that the animals induced to Parkinsonism had lower arterial pressure (AP) and heart rate HR) compared to control animals. We showed that after activation of the baroreceptors by phenylephrine (Phe) and sodium nitroprusside (SNP), the baroreflex sensitivity index was not changed between the groups. However, there was a greater increase in the AP when stimulated with Phe and greater tachycardia when stimulated with SNP in 6-OHDA animals. After activation of the peripheral chemoreceptors through KCN injection (cytotoxic hypoxia), there was a higher increase in pressor and bradycardic response in injured animals with bilateral 6-OHDA. These changes in the cardiovascular reflexes may be important adjustments mechanisms to maintain the cerebral blood flow in those animals, and may be a result of denervation supersensitivity to catecholamines in autonomic targets.

We aimed to investigate the cardiovascular changes in an animal model of Parkinsonism. Adult male Wistar rats were anesthetized to receive bilateral microinfusion of 6-hydroxy dopamine (6-OHDA) or ...

The objective of this study was to investigate the effect of swimming on the cardiac morphological and hemodynamic changes in rats with hypertension induced by L-NAME.. Forty Wistar rats were divided into four groups: sedentary control (SC), trained control (TC), sedentary with L-NAME (LS) and trained with LNAME (LT). The animals in the training groups performed swimming lasting up to 60 min for four weeks. Animals in the L-NAME groups received 20 mg.kg-1 during four weeks. The results showed that animals in the LS group had higher mean arterial pressure (136.6±5.1 mmHg) compared to CS (107.1±1.8 mmHg). The LT group showed significant reductions in mean arterial pressure compared to LS (121.2±1.4 and 136.6±5.1 mmHg, respectively). On the other hand, the LT group animals still remained hypertensive compared to CT group (121.0±1.4 and 107.1±1.8 mmHg respectively). In relation to HR, was observed resting bradycardia for the trained animals. The groups CS and CT showed no changes in relative and absolute weight of the heart. An increase in the absolute weight of the heart to the LS group compared to the CS and also observed an increase in the relative and absolute weight for the LT group compared to CT. Histologic analysis showed that exercise training can reduce the amount of damage caused by chronic administration of L-NAME. In conclusion, we observed that mild exercise was effective in reducing mean arterial pressure in hypertensive rats. Additionally, exercise training did not induced cardiac hypertrophy, but the L-NAME increase the absolute and relative weight of the heart in sedentary and trained rats.

A menopausa é resultante da expressiva redução da produção dos hormônios ovarianos que ocorre durante o climatério. Sabe-se que esses hormônios, principalmente os estrogênios, influenciam positivamente a regulação autonômica cardiovascular, inclusive por meio de suas ações sobre o sistema renina-angiotensina-aldosterona (SRAA). No entanto, em mulheres que mesmo antes do climatério apresentam doença cardiovascular, caso da hipertensão arterial sistêmica (HAS), as consequências sobre a regulação autonômica cardiovascular podem ser ainda mais graves. Nesse caso, a prescrição farmacológica, inclusive com drogas com ação sobre o SRAA, associada às mudanças nos hábitos de vida, como a prática regular de exercícios físicos aeróbios, é frequentemente indicada e apresenta resultados hemodinâmicos bastante animadores. Contudo, desconhecemos os efeitos da associação do tratamento farmacológico e treinamento físico aeróbio sobre o controle autonômico cardiovascular dessas mulheres hipertensas, antes e após a menopausa. Objetivos: investigar em mulheres hipertensas, tanto em idade reprodutiva quanto após a menopausa fisiológica, os efeitos do tratamento farmacológico com Maleato de Enalapril e Losartan associados ao treinamento físico aeróbio sobre a hemodinâmica e sobre o controle autonômico cardiovascular. Métodos: 60 mulheres hipertensas e 60 mulheres normotensas sedentárias foram divididas em quatro grupos; grupo pré-climatério (30 - 45 anos; N=30) e grupo pós-menopausa (55 - 65 anos; N=30). Todos os grupos foram submetidos aos seguintes procedimentos experimentais: lipidograma completo; análise antropométrica; avaliação da capacidade cardiopulmonar por meio do teste ergoespirométrico; avaliação do controle autonômico cardiovascular por meio da análise da sensibilidade barorreflexa e da variabilidade da frequência cardíaca (VFC) e da pressão arterial (VPA). Adicionalmente, os grupos de mulheres hipertensas (pré-climatério e pós-menopausa) foram submetidas a um protocolo de treinamento físico aeróbio, supervisionado e monitorado por 16 semanas, 3 vezes na semana, com duração de 70 minutos por sessão. Resultados: O controle da pressão arterial (PA) por meio do tratamento farmacológico, não normalizou os parâmetros autonômicos cardiovasculares, quando comparado com as mulheres normotensas, tanto antes quanto após a menopausa. Contudo, o grupo hipertenso pré-climatério apresentou um predomínio do componente autonômico vagal no balanço da modulação autonômica cardíaca. Por outro lado, o treinamento físico aeróbio promoveu efeitos positivos sobre a hemodinâmica reduzindo a PA, frequência cardíaca e aumentando o VO2pico, e sobre a modulação autonômica cardiovascular aumentando parâmetros que representam a modulação vagal sob diferentes métodos de análises da VFC e reduziu as oscilações de LF da VPA. Conclusão: A terapia farmacológica é efetiva no controle da PA, porém não normalizou a modulação autonômica cardiovascular em mulheres hipertensas, tanto antes como após a menopausa. O treinamento físico aeróbio foi importante para reduzir a influência simpática sobre os vasos, e promoveu efeitos positivos sobre a VFC. Menopause results from the significant reduction in the ovarian hormones production that occurs during the climacteric. These hormones, especially estrogens, are known to positively influence cardiovascular autonomic regulation, including through their actions on the renin-angiotensin-aldosterone system (RAAS). However, in women who even before the climacteric have cardiovascular disease, in the case of systemic arterial hypertension (SAH), the consequences on cardiovascular autonomic regulation can be even more serious. In this case, the pharmacological prescription, including drugs with action on the RAAS, associated with changes in lifestyle, such as the regular practice of aerobic physical exercises, is often indicated and presents very encouraging hemodynamic results. However, we are not aware of the effects of the association of pharmacological treatment and aerobic physical training on the cardiovascular autonomic control of these hypertensive women, before and after menopause. Aim: to investigate in women with SAH, both at reproductive age and after physiological menopause, the effects of pharmacological treatment with Enalapril Maleate and Losartan associated with aerobic physical training on hemodynamics and cardiovascular autonomic control. Methods: 60 hypertensive women and 60 normotensive sedentary women were divided into four groups; pre-climacteric group (30 - 45 years; N=30) and post-menopausal group (55 - 65 years; N=30). All groups underwent the following experimental procedures: co lipid profile dosage; anthropometric analysis; assessment of cardiopulmonary capacity through the ergospirometric test; assessment of cardiovascular autonomic control by analyzing baroreflex sensitivity and heart rate (HRV) and blood pressure (BPV) variability. Additionally, the groups of women with SAH (pre-climacteric and post-menopausal) underwent a protocol of aerobic physical training, supervised, and monitored for 16 weeks, 3 times a week, lasting 70 minutes per session. Results: Blood pressure (BP) control through pharmacological treatment did not normalize cardiovascular autonomic parameters, when compared with normotensive women, both before and after menopause. However, the pre-climacteric hypertensive group showed a predominance of the vagal autonomic component in the balance of cardiac autonomic modulation. Aerobic physical training promoted positive effects on hemodynamics by reducing BP, heart rate and increasing VO2peak, and on cardiovascular autonomic modulation by increasing parameters that represent vagal modulation under different methods of HRV analysis and reducing LF of the BPV. Conclusion: Pharmacological therapy is effective in controlling BP, but it did not normalize cardiovascular autonomic modulation in women with SAH, both before and after menopause. Aerobic physical training was important to reduce the sympathetic influence on the vessels and promoted positive effects on HRV.

Maresina-1 (MaR1) é um mediador lipídico pró-resolução (SPM) com efeito anti-inflamatório potente. Assim, o objetivo geral dessa tese foi avaliar o efeito da Maresina-1 em silenciar nociceptores, podendendo assim, atuar no controle da dor inflamatória e como alternativa não hormonal para o tratamento da endometriose. Espcificamente, os objetivos dessa tese foram 1) avaliar o efeito analgésico e anti-inflamatório Maresina-1 em modelo de dor e inflamação induzidos pelo adjuvante completo de Freund (CFA) e carragenina; 2) padronizar um modelo não cirúrgico de dor associada à endometriose; 3) avaliar o efeito da MaR1 no modelo de endometriose. Ademais, essa tese oferece revisões sobre o efeito analgésico dos SPMs e sobre importância da capsaicina e TRPV1 na modulação da dor e inflamação. No primeiro trabalho, demonstramos que a MaR1 reduziu a hiperalgesia mecânica e térmica induzida por carragenina e CFA e o recrutamento de neutrófilos e macrófagos próximo a fibras CGRP+ na pele da pata. Além disso, a MaR1 reduziu ativação das células da glia e produção de IL-1β e TNF-α e na medula espinal. No gânglio da raiz dorsal (DRG), MaR1 reduziu a expressão do RNAm de Nav1.8 e Trpv1, e a liberação de CGRP. Os efeitos analgésicos e anti-inflamatórios duradouros da MaR1 (5 dias com um único tratamento) sugerem que os SPMs se tornar uma nova classe de drogas importantes para o tratamento da dor inflamatória. Para o segundo trabalho, nos propusemos a estabelecer um modelo não cirúrgico de dor associada à endometriose em camundongos. A endometriose é uma doença inflamatória que afeta aproximadamente 10% das mulheres e que cursa com dor pélvica ou abdominal debilitante. Além de requererem cirurgia, os modelos existentes não induzem a dor espontânea, que é o principal sintoma de pacientes com endometriose. A endometriose foi induzida não cirurgicamente pela injeção intraperitoneal de fragmentos dissociados de útero de camundongo doador em camundongo receptor. Nosso modelo produz mudanças neuronais e comportamentais (como presença de dor evocada e dor espontânea) que mimetizam a doença em mulheres. Esse modelo também responde a fármacos clinicamente ativos (letrozol e danazol) e pode ser usado, portanto, para avaliar o efeito de novas terapias. Dessa forma, o próximo objetivo foi investigar o efeito da MaR1 no modelo de endometriose. O tratamento com a MaR1 não só reduz a dor abdominal, mas também diminuiu a porcentagem de camundongos com lesões e o tamanho das lesões. Parte desse efeito está relacionado com a inibição da ativação de neurônios CGRP+, como observado pela diminuição da colocalização de CGRP com NF-κB por imunofluorescência no DRG. Em conclusão, demonstramos que a MaR1 apresenta um efeito analgésico duradouro (5 dias após um único tratamento) no modelo de dor induzida por CFA que está relacionado a sua capacidade em bloquear a ativação de nociceptores TRPV1+ e a liberação do neuropeptídeo CGRP. Ademais, padronizamos um modelo não-cirúrgico de dor associada à endometriose que é sensível a fármacos clinicamente ativos, como letrozol e danazol. Demonstramos também que a MaR1 reduz não apenas a dor abdominal, mas também a porcentagem de camundongos com lesões e o tamanho das lesões endometrióticas. Parte desse efeito está relacionado com a habilidade da MaR1 em reduzir a ativação de nociceptores CGRP+. Esses dados indicam que a MaR1 é uma molécula promissora para o tratamento de doenças inflamatórias, como a endometriose. Maresin-1 (MaR1) is a specialized pro-resolving lipid mediator with potent anti-inflammatory activity. We aimed at determining whether MaR1 can silence nociceptors to reduce inflammatory pain and emerge as a non-hormonal alternative for endometriosis treatment. Specifically, we aimed at 1) address the analgesic and anti-inflammatory effect of MaR1 in the carrageenan or complete Freund’s Adjuvant (CFA) models; 2) set out a non-surgical model of endometriosis-associated pain, and 3) investigate the effect of MaR1 in the endometriosis model. This thesis also brings two reviews about 1) the analgesic (without immunosuppressive) effects of SPMs and 2) the role of capsaicin and TRPV1 in modulating inflammatory response. In the first original research work, MaR1 reduced carrageenan- and CFA-induced mechanical and thermal hyperalgesia, and neutrophil and macrophage recruitment proximal to CGRP+ fibers in the paw skin. Moreover, MaR1 reduced spinal cord glial cells activation and TNF-α and IL-1β production. In the DRG, MaR1 reduced CFA-induced Nav1.8 and Trpv1 mRNA expression, calcium influx in TRPV1+ nociceptors, and capsaicin-induced release of CGRP by DRG neurons. The enduring analgesic and anti-inflammatory of MaR1 suggest that the SPMs may become a new class of drugs important for the treatment of inflammatory pain. For the second original research work, we set out to establish a murine model of endometriosis-associated pain. Endometriosis is an estrogen-dependent inflammatory disease that affects approximately 10% of women. Debilitating pelvic or abdominal pain is one of its major clinical features. Current animal models of endometriosis-associated pain require surgery either to implant tissue or to remove the ovaries and do not induce spontaneous pain, which is the primary symptom of patients with chronic pain, including endometriosis. Endometriosis was induced non-surgically by injecting intraperitoneally a dissociated uterine horn into a recipient mouse. In conclusion, we have established a model of endometriosis-associated pain that recapitulates neuronal and behavioral changes in women with endometriosis. In addition, this model responds to clinically active drugs and can be used, therefore, to screen for novel therapies. Therefore, we next addressed whether MaR1 reduces endometriosis-associated pain. We showed that MaR1 reduces the activation of a subpopulation of nociceptors that are responsible for producing neuropeptides such as CGRP. We also demonstrated that MaR1 not only reduced pain but also the number of mice with endometriotic lesions, indicating it is a promising drug for the treatment of endometriosis. In conclusion, we demonstrated that MaR1 presents a long-lasting analgesic effect (5 days after a single treatment) in the CFA-induced pain model which is related to the its ability to block the activation of TRPV1+ nociceptors and the release of the neuropeptide CGRP. Furthermore, we set out a non-surgical pain model associated with endometriosis that is sensitive to clinically active drugs such as letrozole and danazol. In terms of new molecules, treatment with MaR1 reduced not only abdominal pain but also the percentage of mice with lesions and lesion size. Part of that effect is related to MaR1's ability to reduce the activation of CGRP+ nociceptors. These data indicate that MaR1 is a promising molecule for the treatment of inflammatory diseases such as endometriosis.

Com o envelhecimento, o risco de doenças cardiovasculares (DCVs) é maior no sexo feminino, devido principalmente à interrupção na produção dos hormônios ovarianos e é intensificado quando em conjunto à obesidade. O estradiol é enfatizado como um agente cardioprotetor e o uso de terapia hormonal tem sido sugerido como estratégia para prevenção de DCVs em mulheres, mas esta conduta permanece controversa quanto ao papel benéfico ou maléfico ao organismo feminino. Dessa forma, o objetivo deste estudo foi de investigar os efeitos da ausência hormonal ovariana e do tratamento com estradiol, sobre parâmetros cardiovasculares, autonômicos e inflamatórios de ratas induzidas a obesidade com glutamato monossódico (MSG). Este trabalho foi realizado em duas partes, no qual foram utilizadas ratas Wistar adultas e divididas em 4 grupos para cada etapa. Parte 1 Grupos: não obesas não ovariectomizadas (CTR SHAM); não obesas ovariectomizadas (CTR OVX); obesas não ovariectomizadas (MSG SHAM); obesas ovariectomizadas (MSG OVX). Parte 2 Grupos: não obesas ovariectomizadas tratadas com óleo de amêndoas (CTR OL); não obesas ovariectomizadas tratadas com estradiol (CTR ESTR); obesas ovariectomizadas tratadas com óleo de amêndoas (MSG OL); obesas ovariectomizadas tratadas com estradiol (MSG ESTR). A obesidade foi induzida pela administração intradérmica de 4 mg/g de peso corporal de MSG ou salina equimolar nos 5 primeiros dias de vida de ratas. Em todos os grupos a OVX ou falsa OVX foram realizadas aos 75 dias de vida das ratas. O tratamento com estradiol (1mg/kg animal) ou seu veículo (óleo de amêndoas) foram realizados durante 8 semanas após a OVX. Após as 8 semanas todas as ratas foram submetidas à cateterização da artéria femoral para a monitorização da pressão arterial média (PAM), frequência cardíaca (FC) e submetidas à eutanásia para a coleta do encéfalo, coração, rins, aorta, útero, tíbia, tecido adiposo retroperitoneal e coleta do plasma. Os resultados da parte 1 do estudo mostraram que as ratas CTR OVX apresentaram maior peso corporal em relação as CTR SHAM. O peso do tecido adiposo retroperitoneal, foi maior nas ratas MGSs OVX e não OVX comparados aos respectivos controles. O peso uterino das ratas OVX tanto MSG e CTR foram menores do que das ratas não OVX. A frequência cardíaca (FC) das ratas MSGs foram maiores comparadas com as CTRs. A análise espectral do intervalo de pulso (IP) no grupo MSG OVX obteve elevação do LF, e diminuição do HF. Em relação ao óxido nítrico (NO), houve aumento nos rins no grupo CTR OVX e diminuição no MSG OVX em relação ao CTR OVX. No entanto, no coração diminuiu no CTR OVX e MSG OVX comparados ao CTR SHAM e MSG SHAM. No plasma, a concentração de NO foi maior no grupo MSG OVX quando comparado ao MSG SHAM. A expresão da enzima iNOS no tecido cardíaco, é elevado no grupo MSG OVX comparado ao CTR OVX e ao MSG SHAM, nos rins está maior no grupo MSG OVX em relação ao grupo CTR OVX e no cérebro a expressão da enzima é maior no grupo MSG OVX comparado ao MSG SHAM. Os resultados das análises da parte 2 do estudo, mostraram que o ganho de peso corporal do grupo MSG ESTR foi menor do que nos grupos CTR ESTR e MSG OL, houve aumento do peso da tecido adiposo retroperitoneal nos animais do grupo MSG OL em relação aos grupos CTR OL e MSG ESTR, sendo que os animais do grupo MSG ESTR apresentaram valor significativo menor de peso da tecido adiposo retroperitoneal em comparação com o grupo MSG OL e CTR ESTR. Os animais obesos tanto tratados com óleo quanto com estradiol apresentaram menor comprimento em relação aos seus grupos controles (CTR OL e CTE ESTR). Houve um aumento no cálculo do índice de Lee nos animais MSGs (MSG OL e MSG ESTR) em relação aos seus controles (CTR OL e CTR ESTR) respectivamente e uma diminuição deste cálculo no grupo obeso tratado com estradiol (MSG ESTR) comparado ao grupo MSG OL. O peso uterino das ratas do grupo CTR ESTR e MSG ESTR foi maior quando comparados aos grupos controles CTR OL e MSG OL. Os resultados da FC mostraram-se elevados no grupo MSG OL quando comparado ao grupo CTR OL. Na análise da modulação autonômica da pressão arterial sistólica (PAS) houve aumento da banda LF no grupo CTR OL em relação aos grupos CTR ESTR e MSG OL. A efetividade do barorreflexo (ALL Bei) foi menor no grupo MSG OL em relação ao CTR OL e o ganho down foi menor no grupo MSG ESTR comparado ao CTR ESTR. O grupo obeso tratado com estradiol diminuiu a concentração plasmática de NO comparado ao grupo controle com estradiol. No rim o grupo MSG ESTR demonstra maior expressão de iNOS no grupo MSG ESTR em relação ao grupo MSG OL, já no cérebro esta elevado nos grupos MSG OL e MSG ESTR, comparados aos seus controles respectivamente e ainda é maior a expressão da enzima iNOS no grupo MSG ESTR comparado ao grupo MSG OL. Pode se concluir que a ausência dos hormônios ovarianos em associação com a obesidade MSG, atuam de forma efetiva em alterações cardiovasculares, autonômica e inflamatória, no entanto, diante da terapia hormonal com estradiol, conclui-se que o tratamento diminui os fatores de risco para desenvolver DCVs, inibindo a ação do SNS regulando os parâmetros cardiovasculares e autonômicos, porém a terapia não reverte o processo inflamatório. With aging, the risk of cardiovascular disease (CVD) is higher in females, mainly due to the interruption in the production of ovarian hormones and is intensified when combined with obesity. Oestradiol is emphasized as a cardioprotective agent and the use of hormone therapy has been suggested as a strategy for preventing CVD in women, but this conduct remains controversial as to the beneficial or harmful role to the female organism. Thus, the objective of this study was to investigate the effects of ovarian hormonal absence and oestradiol treatment on cardiovascular, autonomic and inflammatory parameters of rats induced obesity with monosodium glutamate (MSG). This work was performed in two parts, in which adult Wistar rats were divided into 4 groups for each stage. Part 1 Groups: non-obese non-ovariectomized (CTR SHAM); non-obese ovariectomized (OVX CTR); obese non-ovariectomized (MSG SHAM); ovariectomized obese women (MSG OVX). Part 2 Groups: ovariectomized nonobese treated with almond oil (CTR OL); ovariectomized ovariectomized women treated with estradiol (CTR ESTR); obese ovariectomized treated with almond oil (MSG OL); ovariectomized obese women treated with estradiol (MSG ESTR). Obesity was induced by the intradermal administration of 4 mg / g body weight of MSG or equimolar saline in the first 5 days of rat life. In all groups the OVX or false OVX were performed at the 75 days of rats life. Treatment with estradiol (1mg / kg animal) or your vehicle (almond oil) was performed for 8 weeks after OVX. After 8 weeks, they underwent femoral artery catheterization for monitoring of mean arterial pressure (MAP), heart rate (HR) and submitted to euthanasia for collection of the brain, heart, kidneys, aorta, uterus, tibia and retroperitoneal fat, and plasma collection. The results from part 1 of the study showed that CTR OVX rats had higher body weight than CTR SHAM. The weight of retroperitoneal fat was higher in rats MGSs OVX and non-OVX compared to the respective controls. The uterine weight of both MSG and CTR OVX rats were lower than non-OVX rats. HR of MSG rats were higher compared to CTRs. The spectral analysis of the pulse interval (PI) in the MSG OVX group obtained elevation of LF, and decrease of HF. In relation to nitric oxide (NO), there was an increase in the kidneys in the OVX CTR group and a decrease in the OVX MSG in relation to the OVX CTR. However, the heart decreased in the CTR OVX and MSG OVX compared to CTR SHAM and MSG SHAM. In plasma, NO concentration was higher in the MSG OVX group when compared to MSG SHAM. Expression of iNOS enzyme in cardiac tissue is elevated in MSG OVX group compared to CTR OVX and MSG SHAM, kidney is elevated in MSG OVX group compared to CTR OVX group and brain expression of enzyme is higher in MSG group OVX compared to MSG SHAM. The results of the analyzes of part 2 of the study showed that the body weight gain of the MSG ESTR group was lower than in the CTR ESTR and MSG OL groups. CTR OL and MSG ESTR, and animals from the MSG ESTR group had a significantly lower value of retroperitoneal fat weight compared to the MSG OL and CTR ESTR group. Obese animals treated with both oil and estradiol were shorter in length than their control groups (CTR OL and CTE ESTR). There was an increase in the Lee index calculation in the MSGs (MSG OL and MSG ESTR) animals compared to their controls (CTR OL and CTR ESTR) respectively and a decrease of this calculation in the estradiol-treated obese group (MSG ESTR) compared to the group. MSG OL. The uterine weight of CTR ESTR and MSG ESTR group rats was higher when compared to CTR OL and MSG OL control groups. HR results were high in the MSG OL group when compared to the CTR OL group. In the analysis of autonomic modulation of systolic blood pressure (SBP) there was an increase in the LF band in the CTR OL group compared to the CTR ESTR and MSG OL groups. Baroreflex effectiveness (ALL Bei) was lower in the MSG OL group compared to CTR OL and down gain was lower in the MSG ESTR group compared to CTR ESTR. The estradiol-treated obese group decreased the NO plasma concentration compared to the estradiol control group. In the kidney the MSG ESTR group demonstrates higher iNOS expression in the MSG ESTR group compared to the MSG OL group, in the brain it is higher in the MSG OL and MSG ESTR groups compared to their controls respectively and the iNOS enzyme expression is higher in the brain. MSG ESTR group compared to MSG OL group. It can be concluded that the absence of ovarian hormones in association with obesity MSG, act effectively in cardiovascular, autonomic and inflammatory changes. develop CVDs, inhibiting the action of the NHS by regulating cardiovascular and autonomic parameters, but therapy does not reverse the inflammatory process.

O câncer de mama é conhecido pela sua alta incidência entre as mulheres no mundo e muito já se conhece sobre os diversos fatores envolvidos no seu surgimento e desenvolvimento. Entre os fatores ambientais, os agrotóxicos vêm ganhando destaque por se mostrarem capazes de atuar na progressão tumoral e gerar quadros de pior prognóstico. Vários mecanismos de lesão ao DNA podem estar envolvidos, mas, pouco se sabe sobre os efeitos dos pesticidas nos níveis plasmáticos de citocinas pró-inflamatórias, hormônios e substâncias envolvidas com o perfil redox, as quais são importantes no contexto da carcinogênese, e, a relação destas substâncias com características prognósticas importantes no câncer de mama. Além disso, fatores relacionados ao estilo de vida das pacientes, como a obesidade, também parecem contribuir para um desfecho de pior prognóstico, principalmente pela existência de um ambiente pró-inflamatório de baixo grau crônico instalado, o qual promove alterações hormonais importantes, como no eixo do cortisol. Assim este trabalho teve como objetivos: avaliar se mulheres portadoras de câncer de mama expostas cronicamente aos agrotóxicos apresentam relação entre desregulação imunoendócrina e desbalanço redox com desfechos de pior prognóstico e, analisar a correlação dos níveis de cortisol sistêmico e parâmetros de pior prognóstico em pacientes com câncer de mama, levando em consideração a categorização das pacientes de acordo com IMC (Índice de Massa Corpórea). A população de estudo foram mulheres com indicativo de câncer de mama atendidas no Hospital do Câncer de Francisco Beltrão, Paraná, Brasil, entre os períodos de 2015 e 2017. Durante a cirurgia para retirada de material para biópsia, no período das 14 às 17 horas, amostras de sangue (10 mL) foram coletadas assim como, das pacientes com diagnóstico positivo para câncer de mama também foram coletadas características clinicopatológicas dos prontuários médicos. Todos os dados foram analisados através do Graph Pad Prism 7.0 e SPSS Statistic 20.0, considerando significância estatística p≤ 0,05. Para demonstrar se há relação dos agrotóxicos com marcadores endócrinos, imunológicos e de estresse oxidativo, iniciamente as pacientes (n=73), que responderam a um questionário sobre exposição direta aos agrotóxicos, foram categorizadas como expostas ou não-expostas. Das amostras de sangue destas pacientes foram dosados: cortisol, citocinas (TNF-α e IL-1β) e o perfil redox (hidroperóxidos, NO e TRAP), para posterior correlação com parâmetros clinicopatológicos. Observamos que os níveis de TNF-α, cortisol e TRAP variaram em relação ao status menopausal das pacientes. Os níveis de TNF-α e TRAP foram aumentados nas pacientes menopausadas não expostas aos pesticidas quando comparadas às expostas (p=0,0443 e p=0,0488, respectivamente). Já nas pacientes na pré-menopausa, estas apresentaram um aumento significativo nos níveis de cortisol no grupo não exposto em relação aos expostos (p=0,0262). Ao contrário, o TRAP nas mulheres na pré-menopausa expostas apresentaram níveis aumentados em comparação com as não expostas (p=0,0006). Além disso, as pacientes não expostas portadoras de tumores de grau 2 apresentaram níveis reduzidos de hidroperóxidos quando comparadas as pacientes expostas do mesmo grau (p=0,0488). Outra diferença observada foi à redução dos níveis de TNF-α nas pacientes expostas quando comparadas as não expostas, em relação a presença de êmbolos angiolinfáticos (p = 0,0128). Em relação a IL-1β, pacientes obesas não expostas apresentaram níveis aumentados em relação às expostas aos pesticidas (p=0,0247). Desta forma, podemos concluir que a exposição crônica aos pesticidas em mulheres com câncer de mama pode contribuir com desfechos de pior prognóstico, em decorrência de desregulação imunoendócrina e redox, influenciada principalmente pelo status menopausal das pacientes. Em seguida, foi analisado se o nível de cortisol plasmático está associado a parâmetros de pior prognóstico quando as pacientes são categorizadas de acordo com o IMC (n=115), sem levar em consideração a exposição aos agrotóxicos. Observou-se que o cortisol sistêmico foi significativamente aumentado nas pacientes com câncer de mama obesas em relação às eutróficas (p=0,051). Pacientes obesas apresentaram cortisol elevado em associação com tumor Luminal B em relação às eutróficas (p=0,0211) e as sobrepesos (p=0,0191), enquanto as sobrepesos apresentaram níveis elevados de cortisol em associação com tumores Triplo Negativo em relação às eutróficas (p=0,0310) e as obesas (p=0,0296). Além disso, a presença de metástase linfonodal, de êmbolos angiolinfáticos e de tumores de alto grau foi relacionada com níveis aumentados de cortisol sérico em pacientes com excesso de peso corporal. Assim, IMC elevado parece ter um efeito negativo sobre o câncer de mama, levando em consideração os níveis de cortisol sérico e parâmetros de pior prognóstico. Breast cancer is known for its high incidence among women in the world and much is already known about the various factors involved in its emergence and development. Among the environmental factors, the pesticides have been gaining prominence because they are able to act in the tumor progression and generate pictures of worse prognosis. Several mechanisms of DNA damage may be involved, but little is known about the effects of pesticides on the plasma levels of proinflammatory cytokines, hormones and substances involved with the redox profile, which are important in the context of carcinogenesis, and the of these substances with important prognostic features in breast cancer. In addition, factors related to patients' lifestyle, such as obesity, also seem to contribute to a worse prognosis outcome, mainly due to the existence of a chronic low-level pro-inflammatory environment, which promotes important hormonal changes, such as cortisol axis. The objective of this study was to evaluate whether women with breast cancer who are chronically exposed to pesticides present a relationship between immunoendocrine dysregulation and redox imbalance with worse prognosis, and to analyze the correlation between systemic cortisol levels and worse prognostic parameters in patients with breast cancer, taking into consideration the categorization of patients according to BMI (Body Mass Index).The study population were women with indicative of breast cancer attended at the Francisco Beltrão Cancer Hospital, Paraná, Brazil, between the periods of 2015 and 2017. During the surgery to remove material for biopsy, from 2 pm to 5 pm, blood samples (10 mL) were collected, as well as the clinicopathological characteristics of the patients’ medical records with positive diagnosis for breast cancer. All data were analyzed using Graph Pad Prism 7.0 and SPSS Statistic 20.0, considering statistical significance p≤0.05. In order to demonstrate the relationship between pesticides with endocrine, immunological and oxidative stress markers, patients (n = 73), who responded to a questionnaire on direct exposure to pesticides, were initially categorized as exposed or non-exposed. Of the blood samples from these patients, cortisol, cytokines (TNF-α and IL-1β) and the redox profile (hydroperoxides, NO and TRAP) were measured for posterior correlation with important clinicopathological parameters in breast cancer. We observed that levels of TNF-α, cortisol and TRAP varied in relation to the menopausal status of the patients. TNF-α and TRAP levels were increased in menopausal patients non-exposed to pesticides when compared to exposed ones (p = 0.0443 and p = 0.0488, respectively). In pre-menopausal patients, these presented a significant increase in cortisol levels in the non-exposed group compared to those exposed (p = 0.0262). In contrast, TRAP in exposed pre-menopausal women had increased levels compared to non-exposed (p = 0.0006). In addition, non-exposed patients with grade 2 tumors had reduced levels of hydroperoxides when compared to exposed patients of the same grade (p = 0.0488). Another difference observed was the reduction of TNF-α levels in exposed patients when compared to non-exposed patients, in relation to the presence of angiolymphatic emboli (p = 0.0128). Regarding IL-1β, obese non-exposed patients presented increased levels in relation to those exposed to pesticides (p = 0.0247). In this way, we can conclude that there is a relation between chronic exposure to pesticides in women with breast cancer and outcomes of worse prognosis, due to immunoendocrine and redox dysregulation, mainly influenced by the menopausal status of the patients. Next, it was analyzed whether the level of plasma cortisol is associated with worse prognostic parameters when the patients are categorized according to the BMI (n = 115), without taking into account exposure to pesticides. It was observed that systemic cortisol was significantly increased in patients with obese breast cancer in relation to eutrophic (p = 0.051). Obese patients had elevated cortisol in association with Luminal B tumor in relation to eutrophic (p = 0.0211) and overweight (p = 0.0191), whereas overweight had high levels of cortisol in association with Negative Triple tumors in relation to eutrophic (p = 0.0310) and obese (p = 0.0296). In addition, the presence of lymph node metastasis, clots and high-grade tumors was related to increased levels of serum cortisol in patients with excess body weight. Thus, high BMI has a negative effect on breast cancer, taking into account serum cortisol levels and worse prognostic parameters.

A depressão é uma doença que acomete grande parte da população mundial e interfere drasticamente na qualidade de vida do indivíduo. Estudos mostram uma conexão entre depressão e doenças cardiovasculares (DCV) em que a depressão pode aumentar a incidência ou ser causa de piora das mesmas. Dessa maneira o uso de fármacos antidepressivos, sendo os inibidores seletivos da recaptação de serotonina os mais utilizados, pode ser benéfico em pacientes depressivos que apresentem algum risco cardiovascular. Dentre eles, o escitalopram (ESC) destaca-se por sua melhor eficácia e segurança. Assim, o presente trabalho teve como objetivo avaliar os efeitos cardiovasculares do tratamento com ESC durante 21 dias em ratos que foram submetidos à separação materna precoce e apresentaram comportamento depressivo-símile. Foram utilizados ratos Wistar (75 dias) de ambos os sexos para o acasalamento e no período do dia pós-natal 2 ao 15 foi realizado o protocolo de separação materna precoce. Aos 60 dias de idade, os filhotes machos passaram pelo teste do nado forçado para verificação do estado depressivo-símile. Os animais que apresentaram o comportamento depressivo-símile receberam salina ou ESC 5 mg/kg, via intraperitoneal, durante 21 dias e foram submetidos ao registro invasivo da pressão arterial para análise dos seguintes parâmetros: valores basais de pressão arterial e frequência cardíaca, variabilidade da frequência cardíaca, variabilidade da pressão sistólica, bloqueio autonômico da frequência cardíaca, avaliação da frequência cardíaca intrínseca, atividade barorreflexo, resposta hemodinâmica a drogas vasoativas e resposta cardiovascular frente ao estresse agudo por restrição. O tratamento com ESC reduziu o parâmetro de muito baixa frequência da VPS (p= 0,0101), indicando que o tratamento com ESC pode exercer efeitos sobre a modulação miogênica vascular, assim como a frequência cardíaca intrínseca relação ao grupo controle (p= 0,0002). Além disso, o tratamento com ESC aumentou o platô inferior (P1) da curva sigmóide da atividade do barorreflexo em relação ao grupo controle (p= 0,0236), associado ao aumento da atividade parassimpática cardíaca durante a atividade do barorreflexo. No entanto, o tratamento com ESC não modificou os demais parâmetros cardiovasculares analisados no presente trabalho. Levando-se em consideração os resultados obtidos pode-se sugerir que o tratamento com ESC em ratos com comportamento depressivo-símile favorece a resposta bradicárdica do barorreflexo e apresenta uma boa segurança para ser utilizado em pacientes depressivos com DCV, mas a redução da frequência cardíaca intrínseca poderia ser prejudicial em caso de pacientes idosos ou com arritmia cardíaca. Depression is a disease that affects a large part of the world's population and drastically interferes with the individual's quality of life. Studies show a connection between depression and cardiovascular disease (CVD) in which depression can increase its incidence. Therefore, the use of antidepressant drugs could be beneficial in depressive patients who present cardiovascular risk, and selective serotonin reuptake inhibitors are the most used in this case. Among them, escitalopram (ESC) stands out for its better effectiveness and safety. Thus, the present study aimed to evaluate the cardiovascular effects of ESC treatment for 21 days in rats that were underwent to early maternal separation and developed depressive-like behavior in adult age. Male and female Wistar rats (75 days) were used for mating and the early maternal separation was performed between the postnatal day 2 to 15. At 60 days of age, the male offspring performed the forced swimming test to verify the depressive-like state. The animals that presented depressive-like behavior received saline or ESC 5 mg/kg, intraperitoneally, for 21 days and were submitted to invasive blood pressure recording to analyze the following parameters: baseline blood pressure and heart rate, heart rate variability, blood pressure variability, autonomic blockade of heart rate, intrinsic heart rate evaluation, baroreflex activity, hemodynamic response to vasoactive drugs and cardiovascular response to acute restraint stress. Treatment with ESC reduced the very low frequency parameter of blood pressure variability (p = 0.0101), indicating that ESC treatment may exert effects on vascular myogenic modulation, as well as the intrinsic heart rate compared to the control group (p = 0.0002). In addition, the ESC treatment increased the lower plateau (P1) of the sigmoid curve of baroreflex activity compared to the control group (p = 0.0236) associated with increased on cardiac parasympathetic activity However, the ESC treatment did not modify the other cardiovascular parameters analyzed in the present study. Considering that, the results suggest that the ESC treatment in rats with depressive-like behavior favors the bradycardic response of the baroreflex and presents a good safety to be used in depressive patients with CVD, but the reduction of the intrinsic heart rate observed could be harm in elderly or patients that present cardiac arrhythmia.

A doença de Parkinson (DP) é uma doença neurodegenerativa progressiva que se manifesta clinicamente após atingir um estágio patológico avançado. Além dos sinais motores, os pacientes com DP apresentam alterações cardiovasculares e autonômicas. Dados recentes mostraram que ratos induzidos ao parkinsonismo pela administração de 6-hidroxidopamina (6-OHDA) na substância negra pars compacta (SNpc) apresentaram menor pressão arterial média (PAM) e frequência cardíaca (FC), e redução da modulação simpática. O núcleo paraventricular do hipotálamo (PVN) é um local importante para o controle autonômico e cardiovascular em situações fisiológicas e patológicas, e as neurotransmissões gabaérgica e glutamatérgica possuem um papel fundamental nesse controle. Desta forma, avaliou-se a participação da neurotransmissão mediada pelos receptores GABAA e NMDA no PVN no controle cardiovascular, autonômico e baroreflexo de ratos induzidos ao parkinsonismo por 6-OHDA. Ratos Wistar foram submetidos a implante de cânulas-guia direcionadas ao PVN e a toxina 6-OHDA ou solução salina estéril foi administrada bilateralmente no SNpc. Após 6 dias, foi realizado o cateterismo da artéria femoral e 24 horas após recuperação do cateterismo, foram realizados os registros dos parâmetros cardiovasculares dos animais não anestesiados. Os valores basais da PAM e FC dos ratos lesados com 6-OHDA (99 ± 3 mmHg, 291 ± 4 bpm) mostraram-se reduzidos quando comparados ao grupo SHAM (111 ± 6 mmHg, 350 ± 4 bpm). A administração de bicuculina (antagonista dos receptores GABAA) no PVN promoveu um aumento na PAM e FC no grupo sham e acentuou os aumentos de PAM e FC no grupo 6-OHDA, bem como acentuou o aumento da modulação simpática no grupo 6-OHDA. A administração de muscimol (agonista dos receptores GABAA) ou NBQX (antagonista dos receptores não-NMDA) no PVN não promoveu alterações cardiovasculares e autonômicas em ambos os grupos. Já a microinjeção bilateral de LY235959 (antagonista dos receptores NMDA) no PVN do grupo sham não induziu alterações cardiovasculares, mas diminuiu a PAM e a FC no grupo 6-OHDA, bem como reduziu a modulação autonômica no grupo 6-OHDA. Os resultados sugerem que as alterações cardiovasculares e autonômicas observadas nos animais 6-OHDA são decorrentes do aumento do tônus gabaérgico no PVN e que através dos receptores glutamatérgicos NMDA no PVN, a neurotransmissão glutamatérgica está envolvida nesta modulação. Do mesmo modo, os resultados sugerem que através de sua ligação aos receptores NMDA, o glutamato promove uma maior ativação do sistema nervoso simpático pelo PVN e participa da manutenção da PAM e da FC em animais induzidos ao parkinsonismo por 6-OHDA. Parkinson's disease (PD) is a progressive neurodegenerative disease that is clinically manifested after reaching an advanced pathological stage. In addition to the motor signals, patients with PD present cardiovascular and autonomic alterations. Recent data have shown that rats induced to parkinsonism by 6-hydroxydopamine (6-OHDA) in the substantia negra pars compacta (SNpc) showed lower mean arterial pressure (MAP) and heart rate (HR), and reduced sympathetic modulation. The paraventricular nucleus of the hypothalamus (PVN) is an important site of autonomic and cardiovascular control in physiological and pathological situations, and gabaergic and glutamatergic neurotransmission play a fundamental role in this control. In this way, the participation of the neurotransmission mediated by the GABAA and NMDA receptors in PVN in the cardiovascular, autonomic and baroreflex control of rats induced to 6-OHDA parkinsonism was evaluated. Male Wistar rats were submitted to implantation of PVN-directed cannulae and the 6-OHDA toxin or sterile saline solution was administered bilaterally in the SNPC. After 6 days, the femoral artery catheterization was performed and 24 hours after surgery recovery, the cardiovascular parameters of non-anesthetized animals were recorded. The baseline values of MAP and HR of 6-OHDA animals (99 ± 3 mmHg, 291 ± 4 bpm) were reduced when compared to the SHAM group (111 ± 6 mmHg, 350 ± 4 bpm). Administration of bicuculline (GABAA receptor antagonist) in PVN promoted an increase in MAP and HR in the sham group and accentuated the increases in MAP and HR in the 6-OHDA group, as well as accentuated the increase in sympathetic modulation in the 6-OHDA group. Administration of muscimol (GABAA receptor agonist) or NBQX (non-NMDA receptor antagonist) in PVN did not promote cardiovascular and autonomic changes in both groups. However, bilateral microinjection of LY235959 (NMDA receptor antagonist) into sham PVN did not induce cardiovascular changes, but decreased MAP and HR in the 6-OHDA group, as well as reduced autonomic modulation in the 6-OHDA group. The results suggest that the cardiovascular and autonomic changes observed in 6-OHDA animals are due to the increase of gabaergic tonus in PVN and that through glutamatergic NMDA receptors in PVN, glutamatergic neurotransmission is involved in this modulation. Likewise, the results suggest that through its binding to NMDA receptors, glutamate promotes a greater activation of the sympathetic nervous system by PVN and participates in the maintenance of MAP and HR in animals induced by 6-OHDA parkinsonism.

Estudos têm demonstrado que o principal mediador do sistema renina angiotensina (SRA), a angiotensina II (Ang II) é um peptídeo chave na regulação da função muscular, estando relacionado com efeitos benéficos ou deletérios nessas células. Assim, abordagens farmacológicas capazes de modular o SRA podem representar potenciais benefícios na estrutura e função muscular. O objetivo do presente trabalho foi avaliar os efeitos do tratamento farmacológico com o inibidor da enzima conversora de angiotensina enalapril (ENA) ou com o antagonista do receptor da angiotensina losartan (LOS) em parâmetros físicos, cardiovasculares, moleculares e epigenéticos em ratos espontaneamente hipertensos (SHR), submetidos ao teste de esforço progressivo em esteira. Foram utilizados ratos espontaneamente hipertensos (SHR) adultos, divididos em três grupos: controle (CTL, água, n=11), enalapril (ENA: 10mg/kg/dia/vo, n=13), losartan (LOS: 10mg/kg/dia/vo, n=15), sendo tal administração realizada por 28 dias consecutivos. Avaliou-se a pressão arterial média (PAM), frequência cardíaca (FC) e a distância percorrida (metros) desses animais em teste progressivo em esteira, a cada 7 dias durante o período de administração das drogas. Após 28 dias de tratamento, parte da amostra (CTL: 5, ENA: 5, LOS:7) permaneceu por 7 dias sem receber tratamento, no intuito de avaliar o efeito tardio ao tratamento. Os animais foram submetidos a eutanásia, sendo coletados o músculo gastrocnêmio, sóleo e o plasma, estes utilizados para realizar análises da capacidade oxidante e antioxidante total e a metilação global do DNA. A ingestão de enalapril modula o peso corporal dos animais, impedindo o ganho de peso ao longo do tratamento, sendo mais evidente após o 14º dia de tratamento (p=0,0222) e reduzindo os níveis de gordura retroperitoneal (p=0,0272). Redução da PAM estatisticamente significativa foi observada após a 4ª semana de tratamento no grupo ENA comparado ao grupo CTL (p

A caquexia do câncer é uma doença caracterizada pela perda progressiva de massa corporal, não envolvendo apenas a musculatura esquelética, mas também a cardíaca. Embora existam relatos de perda de massa muscular cardíaca em ratos portadores de tumor, até o momento existem poucas relações entre estresse oxidativo e caquexia cardíaca, sendo que, diversas estratégias vem sendo estudadas afim de reverter o quadro da caquexia cardíaca. Desde o século passado a creatina passou a ser utilizada como suplementação em diversas doenças cardiovasculares e, apresentou efeitos promissores. Entretanto, ainda não se descreveu o efeito desta suplementação sobre a caquexia cardíaca induzida pelo câncer. Por essa razão, o objetivo desse trabalho foi investigar o efeito da suplementação com creatina durante o desenvolvimento tumoral em animais portadores da forma sólida do tumor de Walker-256. Para isso, ratos Wistar machos foram divididos em 5 grupos: controle (C), tumor (T – inoculados com 8,0x107 células viáveis em 300µL de PBS, intramuscularmente no flanco direito), tumor suplementado com creatina (TCr – inoculação do tumor + creatina diluída em água mineral – 8g/L). Os animais do grupo T e TCr foram acompanhados em dois tempos experimentais distintos, 5 e 10 dias, dessa forma, os 5 grupos experimentais foram: C, T5 e TCr5 (acompanhados durante 5 dias) e T10 e TCr10 (acompanhados durante 10 dias). Os animais foram pesados no início e ao final de cada período experimental e a ingestão alimentar e de água foi controlada a cada 2 dias. Antes da inoculação do tumor e início com a suplementação e antes da eutanásia, os animais foram avaliados através do exame de eletrocardiograma. No dia da eutanásia, os animais foram pesados, anestesiados, e o sangue foi coletado pela veia porta, em seguida, o coração, este separado entre ventrículos e átrios – ambos em direito e esquerdo –, o músculo gastrocnêmio e o tumor. A perda de peso corporal foi calculada através da diferença entre o peso final e o peso inicial, assim como a perda de musculatura esquelética, representada pelo músculo gastrocnêmio, e cardíaca em ambos ventrículos. A amostra de sangue foi utilizada para realização do hemograma, quantificação de glicose, AOPP, malondialdeido e TNF-α. Para o coração, quantificou-se creatina, creatinofosfato e creatina livre, apresentando aumento da concentração nos grupos suplementados, espessura da parede ventrícular por morfometria, perfil elétrico por eletrocardiograma, quantificação de cálcio no tecido, atividades proteassomal e de calpainas, peroxidação lipídica, proteínas carboniladas e produtos reativos ao ácido tiobarbitúrico. Os animais apresentaram perda de peso progressiva durante o desenvolvimento tumoral e diminuição na ingestão alimentar. Ao analisar o sangue, observou-se redução no número de hemácias, aumento de reticulócitos, plaquetas e leucócitos. A glicose sistêmica também reduziu nos animais com tumor, e o estresse oxidativo (lipoperoxidação e carbonilação de proteínas) e a inflamação sistêmica (TNF-α) aumentaram. Ao comparar essas análises com os animais, da qual, ofertou-se a suplementação com creatina, os mesmos parâmetros são semelhantes ao grupo controle. As análises cardíacas nos animais com tumor revelaram diminuição da espessura da parede do ventrículo direito, resultando em ineficácia da repolarização ventricular, aumento do batimento cardíaco e inversão da onda T. Assim, foi capaz de observar aumento da concentração de cálcio no tecido cardíaco e da atividade da calpaina no ventrículo direito. Em teste de estresse oxidativo, ocorreu aumento peroxidação lipídica, carbonilação de proteínas e de malondialdeído nos animais com tumor. Mais uma vez, a suplementação de creatina foi capaz de impedir que algumas alterações ocorressem. Apesar do desenvolvimento tumoral ser capaz de apresentar mudanças importantes no tecido cardíaco, a suplementação com creatina foi capaz de reverter quase todos os parâmetros analisados, apresentando efeitos protetores. Cancer cachexia is a disease characterized by progressive loss of body mass, involving not not only skeletal muscle, but also cardiac proteins. Although it was related the presence of cardiac muscle mass loss in tumor-bearing rats, there are few researches relating oxidative stress with cardiac cachexia and several strategies have been studied to reverse the cancer-induced cardiac atrophy. Since last century, creatine has been used as supplementation in several cardiovascular diseases and presented promising effects. However, the supplementation effect on cancer-induced cardiac cachexia has not yet been described. Therefore, the objective of this work was to investigate the effect of creatine supplementation during tumor development in animals with Walker-256 tumor solid form. For this, male albino Wistar rats were divided into 5 groups: control (C), tumor (T – 8,0x107 viables cells in 300µL PBS, inoculated intramuscularly in the right flank) and tumor/creatine (TCr – tumor implantation and supplemented with creatine, diluted in mineral water – 8g/L) for 5 and 10 days. Animals of T and TCr groups were followed in two different experimental times, 5 and 10 days, so, the 5 experimental groups were: C, T5 and TCr5 (followed for 5 days) and T10 and TCr10 (followed for 10 days). Animals were weighed in the beginning and in the end of each experimental period, food and water intake were monitored every 2 days. Before tumor inoculation and initiation of creatine supplementation and one day before euthanasia, the animals were evaluated using the electrocardiogram. In the euthanasia day, the animals were weighed, anesthetized and blood were collected through the portal vein, then the heart was removed (and separated into left and right ventricles) and the gastrocnemius muscle and tumor were also collected. Loss of body weight was calculated, as well as loss of skeletal muscle, and cardiac, in the two ventricles. Blood samples were used for complete blood count and quantification of glucose, advanced oxidative protein products (AOPP), malondialdehyde and TNF-α. For heart analysis, creatine, creatine phosphate and free creatine was quantified, as well as the wall thickness of the ventricles, electrical profile, tissue calcium quantification, proteasomal activity, lipid peroxidation, carbonylated proteins and thiobarbituric acid reactive products. Animals showed progressive weight loss during tumor development and decrease in food intake. When blood was analyzed, reduction in the number of available red blood cells was observed and increase in reticulocytes, platelets and leukocytes. Systemic glucose levels decreased and oxidative stress and TNF-α levels increased. Animals that received creatine supplementation demonstrated similarity with control in several parameters. The cardiac analysis in tumor animals demonstrated reduction in the thickness of the wall in right ventricle, resulting in ineffectiveness of ventricular repolarization, increased heart rate and inversion of T wave. Furthermore, it was also observed an increase of calcium concentration in cardiac tissue and in the calpain activity in right ventricle. Oxidative stress analyses showed increased lipid peroxidation, carbonylation of proteins and malondialdehyde levels. Creatine supplementation was able to prevent most of these changes. Although tumor development was able to induce significant alterations in cardiac tissue, the creatine supplementation was able to reverse almost all analysed parameters, presenting protective effect.

A doença de Parkinson (DP) é uma condição neurodegenerativa caracterizada pela perda de neurônios dopaminérgicos na substância negra pars compacta (SNPc). Além dos sinais motores, há disfunções cardiovasculares, como hipotensão arterial postural, hipotensão arterial pós-prandial e labilidade da pressão arterial. Estudos mostram que o exercício físico diminui o risco de desenvolver a DP, assim como retarda sua progressão. Durante o treinamento físico, há um acréscimo na produção periférica de óxido nítrico (NO), o qual modula as adaptações cardiovasculares ao exercício. Contudo, nenhum estudo avaliou os efeitos do exercício sobre os níveis periféricos de NO e os seus efeitos modulatórios nas disfunções cardiovasculares de indivíduos com DP. Este trabalho foi aprovado pelo Comitê de Ética no Uso de Animais da Universidade Estadual de Londrina (nº do processo: 19653.2016.42). Animais treinados (T) ou sedentários (S) foram submetidos a cirurgia estereotáxica para microinfusão bilateral de 6-hidroxidopamina (6-OHDA) ou solução veículo (grupo Sham). Após 6 dias, os animais foram cateterizados (artéria e veia femoral) para registro dos parâmetros basais (24h após a cateterização), seguido da inibição da óxido nítrico sintase (NOS) pelo Nw-nitro-arginina-metil-ester (L-NAME, 10mg/kg ? i.v). A dosagem de nitrito foi realizada na aorta, coração, rim, adrenal e plasma por meio da reação colorimétrica de Griess. Após 4 semanas de natação, houve bradicardia de repouso nos grupos treinados (6-OHDA T e Sham T). O NO estava aumentado na aorta do grupo 6-OHDA S e diminuído nos animais 6-OHDA T. No coração, NO estava aumentado para Sham T quando comparado ao grupo sedentário e diminuído no grupo 6-OHDA T em relação aos animais 6-OHDA S e Sham T. No rim, o NO estava diminuído em 6-OHDA S e Sham T quando comparados ao Sham S, e nas glândulas adrenais, houve diminuição no grupo 6-OHDA T em relação ao 6-OHDA S. A administração de L-NAME promoveu menor aumento da pressão arterial média nos grupos 6-OHDA. As reduções na frequência cardíaca foram melhoradas devido ao treinamento físico. O grupo 6-OHDA S teve menor variância na pressão arterial sistólica, a qual não foi alterada pelo exercício. Nossos dados indicam que o exercício físico não promoveu proteção em relação a diminuição de dopamina no estriado e que as alterações nas respostas ao L-NAME na pressão arterial foram decorrentes da lesão e na frequência cardíaca parecem ter sido devido ao treinamento físico. Além disso, nossos dados mostram alterações no NO periférico o qual parece ser modulado diferentemente pelo exercício e o parkinsonismo. Parkinson's disease (PD) is a neurodegenerative condition characterized by the loss of dopaminergic neurons in the substancia nigra pars compacta (SNPc). In addition to motor signals, there are cardiovascular dysfunction as postural and postprandial arterial hypotension and blood pressure lability. Studies have shown that physical exercise decreases risk of developing PD as well as slowing its progression. During aerobic exercise, there is an increased production of nitric oxide (NO), which modulates the cardiovascular adaptations to exercise. However, no study has evaluated the effects of exercise on peripheral NO levels and its modulatory effects on cardiovascular dysfunctions of subjects with PD. This study was approved by the Ethics Committee of the State University of Londrina (process number: 19653.2016.42). Trained (T) or sedentary (S) animals underwent stereotactic surgery for bilateral 6-hydroxydopamine (6-OHDA) or vehicle microinfusion (Sham group). After 6 days, the animals were catheterized (femoral artery and femoral vein) for baseline parameters (24 hours after catheterization), followed by inhibition of nitric oxid synthase (NOS) by Nw-nitro-arginine-methyl ester (L-NAME, 10mg / kg - i.v). Nitrite concentration was performed in the aorta, heart, kidney, adrenal and plasma through Griess colorimetric reaction. After 4 weeks of swimming, trained animals presented resting bradycardia (6-OHDA T and Sham T). NO was increased in the aorta of 6-OHDA S, and decreased in 6-OHDA T animals. In the heart, NO was increased in Sham T compared to sedentary and decreased in 6-OHDA T relative to 6-OHDA S and Sham T animals. At the kidney, NO decrease in 6-OHDA S and Sham T when compared to Sham S and, in adrenal gland, there was a decrease in 6-OHDA T in relation to 6-OHDA S. Administration of L-NAME promoted a lower mean arterial pressure increase in the 6-OHDA groups. Reductions in heart rate were improved due to physical training. The 6-OHDA S group had lower variance in systolic blood pressure, which was not altered by exercise. Our data indicate that exercise did not promote protection against the decrease in dopamine in the striatum and that changes in L-NAME responses in blood pressure were due to injury and in heart rate appear to have been due to physical training. In addition, our data show alterations in peripheral NO which appears to be modulated differently by exercise and parkinsonism.

Obesidade e desordens hormonais na tireoide são muito comuns na população e ambos são considerados um problema de saúde pública, visto que esses distúrbios metabólicos agravam o risco de infertilidade, por alteração no perfil hormonal e anormalidades nos parâmetros funcionais do trato genital masculino. Assim, o objetivo desse estudo foi avaliar o impacto das desordens dos hormônios tireoidianos (hipotireoidismo e hipertireoidismo) em camundongos alimentados com dieta padrão ou dieta hiperlipídica sobre a morfologia e função de testículos e epidídimos. Para isso, camundongos C57BL/6 adultos machos foram distribuídos em 6 grupos experimentais alimentados com dieta padrão (DP) ou hiperlipídica (DHL) durante 12 semanas. Após ambos grupos foram tratados com salina (DP e DHL), PTU (droga antitireoidiana propiltiouracil – DP+PTU e DHL+PTU) ou T3 (hormônio tri-iodotironina – DP+T3 e DHL+T3) durante mais 30 dias. Ao final os animais foram anestesiados e submetidos à eutanásia. O soro do sangue foi utilizado para dosagem de hormônio luteinizante (LH), folículo estimulante (FSH), testosterona e T4 livre. O testículo e epidídimo foram removidos para dosagem do perfil inflamatório de citocinas (TNFIL1 e IL10) e a quantidade de macrófagos (NAG) e de neutrófilos (MPO), além da contagem espermática, análise histopatológica, morfométrica, estereológica e de dinâmica espermatogênica, somado às análises de estresse oxidativo. Os espermatozoides do ducto deferente foram retirados para análise de morfologia espermática. O peso da próstata e do fígado aumentaram nos grupos obeso (DHL+salina) e hipotireoideo obeso (DHL+PTU). O peso da vesícula cheia diminuiu no grupo hipertireoideo obeso (DHL+T3) e da vesícula vazia aumentou no grupo DHL+PTU. A eficácia na indução do hipotireoidismo e hipertireoidismo foi confirmada pela expressão pituitária de mRNA TSH, na qual ficou aumentada na condição de hipotireoidismo e reduzida na condição de hipertireoidismo independente da dieta; enquanto que a eficácia na indução da obesidade foi confirmada com o aumento significativo do peso dos animais alimentados com DHL+salina e DHL+PTU. A dosagem de LH ficou aumentada no grupo hipertireoideo (DP+T3); enquanto, a dosagem de T4 livre diminuiu nos grupos hipo (DP+PTU e DHL+PTU) e hipertireoideo (DP+T3 e DHL+T3), independente da dieta. A contagem de espermátides e a produção diária de espermatozoides (PDE) no testículo aumentaram no grupo DHL+salina e diminuíram no grupo DP+T3 comparado aos demais grupos. A altura do epitélio germinativo aumentou nos grupos DHL+PTU e DHL+T3. A estereologia epididimária da cabeça teve aumento de epitélio no grupo DP+PTU e diminuição na região intersticial do grupo DHL+T3; na cauda teve aumento na região intersticial do grupo DP+T3. A dosagem das enzimas NAG (macrófago) e MPO (neutrófilo) não tiveram resultado significativo entre os grupos. A dosagem de IL1 no testículo aumentou no grupo DHL+T3 e diminuiu no grupo DHL+salina comparado ao controle (DP+salina), enquanto que as demais citocinas (TNF e IL10) não tiveram resultados significativos. A peroxidação lipídica aumentou no grupo DP+PTU comparado ao controle. Com isso, podemos concluir que tanto a dieta hiperlipídica como a desregulação dos hormônios tireoidianos impactaram o tecido testicular e epididimário, porém o hipertireoidismo e a obesidade associados ou isolados prejudicaram com maior intensidade os parâmetros espermáticos. Obesity and hormonal disorders in the thyroid are very common in the population and both are considered a public health problem, since these metabolic disorders aggravate the risk of infertility, altered hormonal profile and abnormalities in the functional parameters of the male genital tract. The objective of this study was to evaluate the impact of thyroid hormone disorders (hypothyroidism and hyperthyroidism) on mice fed a standard diet or a high-fat diet on the morphology and function of the testis and epididymis. For this, male C57BL/6 mice were distributed in 6 experimental groups fed standard diet (SD) or high-fat diet (HFD) for 12 weeks. After, both groups were treated with saline (SD+saline and HFD+saline), PTU (antithyroid drug propylthiouracil - SD+PTU and HFD+PTU) or T3 (hormone triiodothyronine - SD+T3 and HFD+T3) for another 30 days. At the end, the animals were anesthetized and submitted to euthanasia. Blood serum was used for dosage of luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone and free T4. The testis and epididymis were removed to quantify the inflammatory profile of cytokines (TNF, IL1 and IL10) and the number of macrophages (NAG) and neutrophils (MPO), as well as sperm count, histopathological, morphometric, stereological and spermatogenic dynamics, added to the analyzes of oxidative stress. The spermatozoa of the vas deferens were removed for analysis of spermatic morphology. The weight of the prostate increased in the obese (HFD+saline) and obese hypothyroid (HFD+PTU) groups. The weight of the full vesicle decreased in the obese hyperthyroid group (HFD+T3) and the empty vesicle increased in the HFD+PTU group. The efficacy in the induction of hypothyroidism and hyperthyroidism was confirmed by the pituitary expression of TSH mRNA, in which it was increased in the condition of hypothyroidism and reduced in the condition of diet-independent hyperthyroidism; while the efficacy in obesity induction was confirmed with a significant increase in the weight of the animals fed HFD+saline and HFD+PTU. The LH dosage was increased in the hyperthyroid group (SD+T3); while the free T4 levels decreased in the hypo (SD+PTU and HFD+ PTU) and hyperthyroid (SD+T3 and HFD+T3) groups, regardless of diet. The spermatid count and daily sperm production (PDE) in the testis increased in the HFD+saline group and decreased in the SD+T3 group compared to the other groups. The height of the germinal epithelium increased in the HFD+PTU and HFD+T3 groups. The epididymal stereology of the head had increased epithelium in the DP + PTU group and decreased in the interstitial region of the DHL + T3 group; in the tail had an increase in the interstitial region of the SD+T3 group. The dosages of the NAG (macrophage) and MPO (neutrophil) enzymes did not show significant results between the groups. The IL1 dosage in the testis increased in the HFD+T3 group and decreased in the HFD+saline group compared to the control (SD+saline), whereas the other cytokines (TNFα and IL10) did not have significant results. Lipid peroxidation increased in the SD+PTU group compared to the control group. With this, we can conclude that both the high-fat diet and the dysregulation of thyroid hormones had an impact on the testicular and epididymal tissue, but associated or isolated hyperthyroidism and obesity affected the sperm parameters more intensely.

O óxido nítrico (NO) no núcleo paraventricular do hipotálamo (PVN) é importante neuromodulador de respostas autonômicas e o treinamento físico assim como a suplementação de L-arginina pode induzir diferentes adaptações ao sistema cardiovascular. O objetivo deste estudo foi avaliar os efeitos da enzima nNOS do PVN na neuromodulação cardiovascular de ratos treinados ou suplementados com L-arginina. Após aprovação da Comissão de Ética no Uso de Animais da Universidade Estadual de Londrina (protocolo: 5185.2014.79), ratos Wistar (220-250g) foram submetidos a sedentarismo ou treinamento de natação, corrida ou suplementação oral com L-arginina (dose: 62,5mg/1ml/rato/dia). Posteriormente foi realizada estereotaxia para implante de cânulas-guia direcionadas ao PVN e cateterismo de artéria e veia femoral para registros cardiovasculares. Outros animais foram destinados à coleta de tecidos para análises de composição corporal, nitrito, expressão gênica e imunohistoquímica. A natação e a corrida proporcionaram menor ganho de peso corporal, diminuição de tecido adiposo perigonadal e retroperitoneal e aumento do peso cardíaco. Apenas animais treinados em esteira apresentaram maior peso de músculos gastrocnêmio e sóleo. Ambos os protocolos de treinamento provocaram: bradicardia em repouso, sem alterações na pressão arterial média (PAM); melhoraram a sensibilidade do barorreflexo espontâneo e modulação da variabilidade da frequência cardiaca (FC), através do aumento do componente parasimpático e diminuição do simpático. A microinjeção bilateral de inibidor seletivo da nNOS (N-Propyl-L-Arginina, 4nmol/100nl) no PVN provocou maior resposta pressórica e taquicárdica em ratos treinados e um déficit no controle autonômico cardiovascular. Ratos treinados apresentaram maior prevalência de neurônios imunoreativos para nNOS no PVN, maior expressão gênica de nNOS e níveis de nitrito no PVN. A suplementação com L-arginina proporcionou efeitos similares ao exercício. Animais suplementados apresentaram bradicardia de repouso sem alterações na PAM e maior resposta pressora e taquicárdica ao N-propyl no PVN. A análise espectral basal de ratos suplementados com L-arginina evidenciou menor modulação simpática para pressão arterial sistólica e FC e melhores índices de barorreflexo espontâneo. A suplementação com L-arginina favoreceu maior ganho máximo de sensibilidade barorreflexa, o qual foi abolido após a inibição da nNOS no PVN. Além disso, a microinjeção bilateral de L-arginina (0,17g/ml/100nL) no PVN de ratos controle melhorou o ganho máximo de sensibilidade barorreflexa. Animais suplementados apresentaram menor peso de tecido adiposo perigonadal e retroperitoneal, maior peso de músculo gastrocnêmio e rins e maiores concentrações de nitrito no PVN, rins e aorta torácica. Em conclusão, o treinamento físico e a suplementação oral de L-arginina modificam o controle autonômico do PVN via nNOS e ambos favorecem a biodisponibilidade de NO que, central e perifericamente pode ser relevante para adaptações cardiovasculares. Nitric oxide (NO) into the paraventricular nucleus of hypothalamus (PVN) is an important neuromodulator of autonomic responses, and physical training as well as L-arginine supplementation may induce different adaptations to the cardiovascular system. Thus, the objective of this study was to evaluate the effects of nNOS enzyme of PVN on cardiovascular neuromodulation in rats trained or supplemented with L-arginine. After approval by the Animal Ethics Committee of the State University of Londrina (protocol: 5185.2014.79), Wistar rats (220-250g) were submitted to sedentarism or swimming training, running or oral supplementation with L-arginine (dose: 62.5mg/1ml/rat/day). Subsequently, implantation of guide cannulae directed to the PVN by stereotactic surgery and catheterization of the femoral artery and vein were performed for cardiovascular records. Other animals were used to collect tissues for analysis of body composition, nitrite levels, gene expression and immunohistochemistry. Swimming and running training provided decreased body weight and adipose tissue perigonadal and retroperitoneal and increased heart weight. Only treadmill animals presented greater weight of muscles gastrocnemius and soleus. Both training protocols resulted in: resting bradycardia, no changes in mean arterial pressure (MAP); improved sensitivity of the spontaneous baroreflex and modulation of heart rate variability (HR), through an increase of the parasympathetic and reduction of the sympathetic component. The bilateral microinjection of nNOS selective inhibitor (N-Propyl-L-Arginine, 4nmol/100nl) in PVN caused a greater pressure and tachycardic response in trained rats and a deficit in the autonomic cardiovascular control. Trained rats showed higher prevalence of immunoreactive neurons for nNOS in PVN, higher nNOS gene expression, and nitrite levels in PVN. L-arginine supplementation provided similar effects to those observed by exercise. Supplemented animals had resting bradycardia without changes in MAP, as well as a greater pressure and tachycardic response to N-Propyl in PVN. Basal spectral analysis of L-arginine supplemented rats, showed lower sympathetic modulation for systolic blood pressure and HR and better indices of spontaneous baroreflex. L-arginine supplementation favored a greater maximum baroreflex sensitivity gain, which was abolished after nNOS inhibition in PVN. In addition, bilateral microinjection of L-arginine (0.17g/ml/100nL) into the PVN of control rats improved the maximum baroreflex sensitivity gain. In supplemented animals there was also lower adipose tissue weight perigonadal and retroperitoneal, higher weight of gastrocnemius muscle and kidneys; higher nitrite levels in PVN, kidneys and thoracic aorta. In conclusion, physical training as well as oral supplementation with L-arginine modifies the autonomic control of PVN via nNOS and both favor the bioavailability of NO, which, centrally and peripherally, may be relevant for cardiovascular adaptations.