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Cochlospermum regium is well-known as "Algodãozinho do cerrado" in folk Brazilian medicine, and is used to fight infections, inflammation and skin disorders.To identify the phytochemical constituents and the effects of the ethanolic extract of C. regium leaves (EECR) on inflammation and pain, and the effects of C. regium gel (GEECR) on wound healing.Animals were treated with EECR (30-300 mg/kg) or GEECR (1.25 and 2.5%) and studies were conducted using carrageenan-induced pleurisy and paw edema tests, formalin-induced pain model, and excision wound model.In total, 25 compounds, including quercitrin, methyl gallate, and 1,2,3,4,6-pentagalloylhexose, with highest detectability were identified. The treatments reduced leukocyte migration, nitric oxide production, protein extravasation, edema, mechanical hyperalgesia, pain in both phases (neurogenic and inflammatory), cold hypersensitivity, and improved wound closure and tissue regeneration.The present findings established the anti-inflammatory, anti-nociceptive, and wound healing potential of the leaves of C. regium, confirming the potential therapeutic effect of this plant.

Pachira aquatica is a species used for medicinal and food purposes and has numerous phytochemicals that may have systemic toxic effects and damage to genetic material. This study aimed to evaluate acute and short-term oral toxicity, as well as genotoxic and clastogenic effects of oil extracted from P. aquatica (PASO) seeds in rats and Drosophila melanogaster. The results obtained with biochemical and hematological analyses did not show significant changes in any evaluated parameters when compared with reference values for the species used in the study. Data from the histopathological analysis corroborated results found in this study. These findings indicate low acute and short-term toxicity following oral PASO exposure in rats under the experimental conditions tested. Tests performed in rats showed that PASO did not present significant genotoxic or clastogenic effects on the cells analyzed with the three doses tested. Treatment with PASO in the offspring of HB crossing, which showed high cytochrome P450 levels, did not exhibit genotoxic activity, as demonstrated by the SMART test. These results suggest that products from the hepatic oil metabolism did not show genotoxicity under the conditions tested. Together, the results indicate that, under the experimental conditions tested, PASO is safe for repeated intake. As PASO exhibited low potential to cause harmful effects on living organisms, our study encourages further research aimed at assessing its pharmacological activity, since it is a widely consumed plant.

Dipyrone is an analgesic and antipyretic drug commonly used in many countries. Although generally not recommended during pregnancy, it is known that many women use dipyrone during the gestational period. In this study, we investigated the endocrine and reproductive effects of dipyrone in female and male offspring rats exposed in utero from gestational days 10–21. Pregnant rats were treated with dipyrone at 25, 75, and 225 mg/kg/day via oral gavage. Developmental landmarks—anogenital index (AGI), number of nipples, vaginal opening, first estrus, and preputial separation—were evaluated in the offspring. Reproductive parameters, including estrous cycle regularity, daily sperm production, weight and histopathology of reproductive organs, steroid hormone levels, and gene expression of selected markers of reproductive function were assessed at adulthood. At the highest dose, dipyrone induced a significant increase in postimplantation losses/fetal death and delayed parturition in dams. Offspring exposed in utero to the highest dose also exhibited significant changes in some early life markers of endocrine disruption, in particular increased AGI in females, indicating a proandrogenic effect, and increased rate of retained nipples in males, indicating an antiandrogenic response. No changes were observed in markers of puberty onset or reproductive parameters at adulthood. These results suggest that exposure to therapeutically relevant doses of dipyrone may induce mild endocrine disruptive effects that can be detected in late pregnancy and early life. Such effects may be relevant considering dipyrone use by pregnant women and the possibility of coexposures with other endocrine disruptors.

Diclofenac is a non-steroidal anti-inflammatory drug widely used by the general population and, although generally contraindicated during pregnancy, it is also used by some pregnant women. This study investigated endocrine, reproductive and behavioral effects of diclofenac in male and female offspring rats exposed in utero from gestational days 10-20. Pregnant rats were treated with diclofenac at doses of 0.2, 1 and 5 mg/kg/day via oral gavage. Anogenital distance (AGD), number of nipples, and developmental landmarks of puberty onset - vaginal opening (VO), first estrus (FE) and preputial separation (PPS) - were evaluated in the offspring. At adulthood, behavioral and reproductive parameters were assessed. Male and female rats were tested in the elevated plus maze test to assess locomotor activity and anxiety-like behaviors, while male rats were also evaluated in the partner preference test. No significant effects were observed on AGD and number of nipples in both males and females. Diclofenac treatment induced an overall delay in developmental landmarks of puberty onset in male and female offspring, which reached statistical significance for PPS at the lowest diclofenac dose. Prenatal exposure to all tested doses abolished the preference of male rats for an estrous female, suggesting an impairment of brain masculinization. No changes were observed on male or female reproductive parameters at adulthood. Overall, our results indicate that prenatal exposure to therapeutically relevant doses of diclofenac may have an impact in the pubertal development of rats and negatively affect male partner preference behavior.

Background: Hemorrhagic cystitis is an inflammatory complication that can be caused by the administration of cyclophosphamide, which is widely used as an antineoplastic agent. In the search for new therapeutic alternatives, probiotics can suppress the inflammatory process and, therefore, can be used to prevent this disease. Objective: Thus, this study aimed to evaluate the effects of using Lactobacillus acidophilus NCFM in the treatment of cyclophosphamide-induced hemorrhagic cystitis in Wistar rats. Methods: Lactobacillus acidophilus NCFM (2x108 CFU) was used in the treatment of cyclophosphamide- induced hemorrhagic cystitis (200 mg/kg, intraperitoneal) in 77 female Wistar rats. Rats were distributed into experimental groups (n = 9): control group (GC), zero control group (GCZ), inflammation group (GI), 24-hour acute treatment groups: 24-hour lactobacilli treatment group (GL24H) and mesna group (GM), and 30-day chronic treatment groups: lactobacilli treatment group (GTL) and mesna+lactobacilli group (GM+L). After treatment, animals were euthanized and biological materials were collected for blood count, biochemical analyses, examination of abnormal sediment elements (EAS), and histopathological analysis. Results: GI results showed development of edema, macroscopic alterations, and signs of bleeding in the bladder; in addition, lesions in the urothelium and hemorrhage were also found. GL24H and GM presented intact urothelium, without inflammatory reaction and hematological or biochemical urine alterations. Conclusion: Therefore, this study demonstrated that L. acidophilus presented uroprotective effect against the action of cyclophosphamide in both the short and long term.

Croton urucurana Baill. is a native Brazilian tree, popularly known as “sangra-d'água” or “sangue-de-dragão,” based on the red resinous sap of the trunk. Its use has been transmitted through generations based on popular tradition that attributes analgesic, anti-inflammatory, and cardioprotective properties to the tree. However, its cardioprotective effects have not yet been scientifically investigated. Thus, the present study investigated the pharmacological response to an ethanol-soluble fraction from the leaves of C. urucurana in Wistar rats exposed to smoking and dyslipidemia, two important cardiovascular risk factors. The extract was evaluated by high-performance liquid chromatography. Wistar rats received a 0.5% cholesterol-enriched diet and were exposed to cigarette smoke (9 cigarettes/day for 10 weeks). During the last 5 weeks, the animals were orally treated with vehicle (negative control group), C. urucurana extract (30, 100, and 300 mg/kg), or simvastatin (2.5 mg/kg) + enalapril (15 mg/kg). One group of rats that was not exposed to these risk factors was also evaluated (basal group). Electrocardiograms and systolic, diastolic, and mean blood pressure were measured. Blood was collected to measure total cholesterol, triglycerides, urea, and creatinine. The heart and kidneys were collected and processed for oxidative status and histopathological evaluation. The phytochemical analysis revealed different classes of flavonoids and condensed tannins. The model induced dyslipidemia and cardiac and renal oxidative stress and increased levels of urea and creatinine in the negative control group. Treatment with the C. urucurana extract (300 mg/kg) and simvastatin + enalapril decreased cholesterol and triglyceride levels. In contrast to simvastatin + enalapril treatment, the C. urucurana extract exerted cardiac and renal antioxidant effects. No alterations of electrocardiograms, blood pressure, or histopathology were observed between groups. These findings indicate that C. urucurana exerts lipid-lowering, renal, and cardioprotective effects against oxidative stress in a preclinical model of multiple risk factors for heart disease.

Ethnopharmacological relevance Talinum paniculatum (Jacq.) Gaertn. (Talinaceae) is a medicinal species that is widely distributed throughout Brazil. Popularly known as “major-gomes,” the species is used in folk medicine for the treatment of cardiovascular disorders. Aim of the study To evaluate the effect of an ethanolic extract of T. paniculatum (EETP) in rats with renovascular hypertension and heart failure and determine its chemical composition. Materials and methods First, EETP was obtained, and its chemical profile was analyzed by LC-DAD-MS. The acute toxicity was evaluated in female Wistar rats. The model of renovascular hypertension was established in male Wistar rats by combining the Goldblatt 2K1C method and intraperitoneal doxorubicin administration for 6 weeks. The animals were then treated daily with EETP (30, 100, and 300 mg/kg) or metoprolol (25 mg/kg) by gavage for 28 days. The negative control group was treated with vehicle (filtered water). The sham group consisted of animals that were not subjected to 2K1C or cardiotoxicity and were treated with vehicle. Renal function was evaluated on days 1, 14, and 28. At the end of treatment, the electrocardiographic profile, blood pressure, and mesenteric vascular reactivity were investigated. Serum urea, creatinine, angiotensin converting enzyme, nitrotyrosine, malondialdehyde, nitrite, aldosterone, and sodium and potassium levels were measured. The heart, aorta artery, liver, and right kidney were collected, weighed, and processed for histopathological analysis. Cardiac chambers also underwent morphometric analysis. Results No signs of toxicity were observed in female Wistar rats. Thirty-two compounds were annotated from EETP, including flavonoids, chlorogenic acids, and saponins. EETP treatment resulted in a significant cardiorenal-protective response, normalizing electrocardiographic and hemodynamic alterations, and preventing ventricle remodeling. These effects were associated with serum antioxidant activity and angiotensin-converting enzyme (ACE) inhibition. Conclusion The present study demonstrated that EETP may exert cardioprotective effects through serum antioxidant activity and ACE inhibition, preventing alterations of hemodynamic and endothelial function, and reducing damage to cardiac structure. Thus, EETP, especially at the 100 and 300 mg/kg doses, may be useful for preventing doxorubicin-induced cardiotoxicity in hypertensive patients.

Although leaves of Anchietea salutaris are used in Brazilian traditional medicine, there is no available data in the literature proving its efficacy and safety. Thus, the aim of the study was to perform a meticulous botanical, phytochemical, toxicological, and pharmacological investigation of A. salutaris in Wistar rats. At first, a morphoanatomical characterization of Anchietea pyrifolia leaves and stems was performed. Then, a purified infusion (ethanol-soluble fraction obtained from A. pyrifolia [ESAP]) was obtained followed by its chemical profile elucidation. Furthermore, an acute toxicity test was performed, and the acute and prolonged diuretic and hypotensive effects were also evaluated in Wistar rats. Finally, the vasodilatory responses of ESAP in mesenteric vascular beds were investigated. The main secondary metabolites identified from ESAP were O-glycosylated flavonoids, chlorogenic acids, and phenylpropanoic acid derivatives. ESAP did not promote any toxic effects in female rats nor increased urinary excretion in male rats after a single exposure. However, ESAP significantly reduced renal elimination of sodium, potassium, and chloride after prolonged treatment. An ESAP highest dose promoted significant acute hypotension without affecting blood pressure levels after prolonged use. Furthermore, its cardiovascular effects seem to be related with the calcium-activated potassium channel activation in resistance vessels.

The main physiological consequence of diabetes mellitus is chronic hyperglycemia. This condition is related to the formation of free radicals including advanced glycation end products (AGES) and to an increase in inflammatory processes. Cochlospermum regium (Schrank) Pilg., part of the Bixaceae family, is a cerrado plant known for its anti-inflammatory effects. The objectives of this study were to analyze the constituent compounds of C. regium roots and to evaluate the antioxidant, antiglycation, antidiabetic, and anticholinergic effects of its hydromethanolic extract through in vitro and in vivo experimental models. The presence of phenols, flavonoids, condensed tannins, and flavonols was analyzed by liquid chromatography - photodiode array (LC/PDA) analysis. Whereas antioxidant activity was investigated via DPPH, ABTS, β-carotene/linoleic acid, and malondialdehyde colorimetric assays. Inhibition of AGEs was examined via a bovine serum albumin system whose glycosylating agent was glucose. Antidiabetic potential was examined in normoglycemic Wistar rats that received glucose overload, in alloxan-induced diabetic rats, and in rats that received a hyperglycemic diet. Disaccharidase inhibition was assessed using in vitro and in vivo methods, as was acetylcholinesterase (AChE) inhibition in brain structures. The hydromethanolic extract (CRHE) possessed a high concentration of phenolic compounds and showed antioxidant effects. The LC-DAD results revealed that CRHE contained a high concentration of phenolic acids and the majority was gallic acid. Treatment with CRHE caused significant inhibition of AGE formation. The oral glucose tolerance test (OGTT) in normoglycemic animals showed a reduction in blood glucose levels after treatment with 100 mg/kg CHRE, accompanied by an increase in hepatic glycogen content. There was also a significant reduction in the fasting glucose levels of alloxan-induced diabetic animals after 7 days of treatment with daily doses of 100 mg/kg. After 14 weeks of hyperglycemic diet, the last four or which were combined with 100 mg/kg CRHE treatment, there was a decrease in blood triglyceride levels. There was also a statistically significant decrease in the enzymatic activity of maltase, lactase and sucrase. The results demonstrate that oral administration of 30 and 100 mg/kg CRHE inhibited AChE activity in different brain structures. Thus, the extract of C. regium showed promising antioxidant, antiglycation, and antidiabetic effects that may be related to its high phenolic content.

Ethnopharmacological relevance Psidium guineense, popularly known as “araca-do-campo”, is used in popular medicine for the treatment of inflammatory diseases. Our research group studied an essential oil obtained from its leaves and reported anti-inflammatory and analgesic properties. However, to date, the anti-inflammatory actions of the leaf extract have not been evaluated although the traditional folk use of this plant has these indications. Aim of study The current study was designed to evaluate the antioxidant and anti-inflammatory effects and toxicity of the hydromethanolic extract of the leaves from P. guineense (HME-PG), as well as to investigate the chemical composition. Materials and methods HME-PG was chemically investigated by Ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). The antioxidant activity was evaluated with 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and malondialdehyde (MDA). Swiss mice were orally (p.o.) pretreated with HME-PG (30, 100 and 300 mg/kg), and after 1 h received carrageenan via paw injection (edema, cold sensitivity and mechanical hyperalgesia were analyzed) or pleural injection (leukocyte migration was analyzed after 4 h) and for nociception using the formalin model. Acute (14 days) and subacute (28 days) toxicity was assessed with female Wistar rats orally treated with 500 and 2000 mg/kg HME-PG. Results HME-PG showed high levels of phenolic and flavonoid compounds. Six compounds were identified based on UHPLC-MS/MS analysis, including gallic acid, quercetin, 3′-formyl-2′,4′,6′-trihydroxy-5′-methyldihydrochalcone, vanillic acid, ursolic acid and corilagin. HME-PG exhibited an IC50 of 48.14 μg/mL in the MDA assay and an IC50 of 45.15 μg/mL in the DPPH test. The treatment with HME-PG (100 and 300 mg/kg) significantly inhibited edema at all time points evaluated, mechanical hyperalgesia after 4 h and the response to cold 3 and 4 h after carrageenan injection and anti-nociceptive effects in both phases of formalin nociception. All oral HME-PG treatments significantly inhibited leukocyte migration and plasma extravasation in the pleurisy model. Toxicity tests did not cause signs of toxicity in the treated animals. Conclusions The present study showed that HME-PG has antioxidant and anti-inflammatory properties, and no toxicity was detected after acute or subacute treatment with HME-PG, showing the possibility for the safe traditional use of P. guineense.

Background: Costus spicatus (Jacq.) Sw. is a medicinal species frequently prescribed for the treatment of cardiovascular diseases. This study aims to evaluate the effects of this species against the development of atherosclerosis. Methods: First, an anatomical study of the C. spicatus leaves was performed. Then, the extract (ESCS) was obtained and submitted to phytochemical analysis. Female rats were treated with a single dose of ESCS (2000 mg/kg) to assess acute toxicity. Other groups of female rats received an atherogenic diet for 60 days. After 30 days, the animals were treated orally with ESCS (30 and 300 mg/kg), rosuvastatin (5 mg/kg), or vehicle once daily for 30 days. Serum lipids oxidized low-density lipoprotein, soluble adhesion molecules, interleukins 1β and 6, and markers of renal and liver function were measured. Renal function, blood pressure, electrocardiography, and vascular reactivity were also evaluated. Arteries, heart, liver, and kidney were also collected to evaluate the tissue redox state and histopathological analysis. Results: Prolonged treatment with ESCS induces significant hypolipidemic and antioxidant effects, that prevent endothelial dysfunction and modulated the local inflammatory process, reducing the evolution of the atherosclerotic disease. Conclusions: This study provides a scientific basis for the popular use of C. spicatus for the treatment of atherosclerosis.

Ethnopharmacology relevance Aleurites moluccana is popularly used for the diseases like ulcers, fever, headache, asthma, conjunctivitis, gonorrhea, inflammation, hepatitis, and rheumatism. The seed, also known as “noz da India”, has been popularly consumed for weight loss purposes but reports of toxicity have been associated with its ingestion. In the literature, there are not enough studies to elucidate its toxicology, so evaluating the general and genetic toxicological of A. moluccana seeds can provide data to ensure their intake. Aim of the study: The objective of the present study was to elucidate the oral toxicity, mutagenicity, genotoxicity and cytotoxicity of A. moluccana seeds in vitro and in vivo assays. Materials and methods The chemical composition of the aqueous extract of A. moluccana seeds (AEAMS) was analyzed in relation to phenolic compounds, tannins, flavonoids and fatty acid. For the in vitro assays, the cytotoxic potential was assessed by the MTS assay whereas the mutagenic potential was assessed by the Ames test. For in vivo assays, was conducted an acute oral toxicity study, with “Up-and-Down Procedure” and repeated dose toxicity with “Repeated Dose 28-Day Oral Toxicity”. To assess genetic damage, mutagenic potential was assessed by the micronucleus test whereas the polychromatic erythrocyte/normochromatic erythrocyte ratio was obtained with bone marrow cells to determine the cytotoxic potential and genotoxic potential was assessed by the comet assay using peripheral blood cells. Results AEAMS did not show cytotoxic and mutagenic potential in vitro. No clinical signs of toxicity were observed in animals after the acute oral toxicity test, suggesting that the LD50 of aqueous extract of A. moluccana seeds > 2000 mg/kg in a single dose by intragastric route. However, in toxicity at repeated doses for 28 days, the doses initially established (250; 500 and 750 mg/kg/day by intragastric route) caused mortality in the animals and the reestablished doses (25, 50 and 100 mg/kg/day by intragastric route) showed no changes in parameters or clinical signs attributed to toxicity. Furthermore, AEAMS also did not show mutagenic, genotoxic and cytotoxic potential in vivo. Conclusions AEAMS did not show cytotoxic, genotoxic and mutagenic potential in vitro and in vivo. And although the AEAMS has an LD50 > 2000 mg/kg, and does not have physiological, biochemical, hematological, histopathological changes or clinical signs related to toxicity when administered in low concentrations and for a short period, in high concentrations and continued use caused toxicity and mortality in Wistar rats. In order to obtain complementary results, is recommended highly that further mid and long-term toxicological studies are investigated, and in no-rodent specie.

Made available in DSpace on 2022-04-28T19:47:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Croton urucurana Baill. is a native Brazilian tree, popularly known as sangra-d'águaor sangue-de-dragão,based on the red resinous sap of the trunk. Its use has been transmitted through generations based on popular tradition that attributes analgesic, anti-inflammatory, and cardioprotective properties to the tree. However, its cardioprotective effects have not yet been scientifically investigated. Thus, the present study investigated the pharmacological response to an ethanol-soluble fraction from the leaves of C. urucurana in Wistar rats exposed to smoking and dyslipidemia, two important cardiovascular risk factors. The extract was evaluated by high-performance liquid chromatography. Wistar rats received a 0.5% cholesterol-enriched diet and were exposed to cigarette smoke (9 cigarettes/day for 10 weeks). During the last 5 weeks, the animals were orally treated with vehicle (negative control group), C. urucurana extract (30, 100, and 300 mg/kg), or simvastatin (2.5 mg/kg) + enalapril (15 mg/kg). One group of rats that was not exposed to these risk factors was also evaluated (basal group). Electrocardiograms and systolic, diastolic, and mean blood pressure were measured. Blood was collected to measure total cholesterol, triglycerides, urea, and creatinine. The heart and kidneys were collected and processed for oxidative status and histopathological evaluation. The phytochemical analysis revealed different classes of flavonoids and condensed tannins. The model induced dyslipidemia and cardiac and renal oxidative stress and increased levels of urea and creatinine in the negative control group. Treatment with the C. urucurana extract (300 mg/kg) and simvastatin + enalapril decreased cholesterol and triglyceride levels. In contrast to simvastatin + enalapril treatment, the C. urucurana extract exerted cardiac and renal antioxidant effects. No alterations of electrocardiograms, blood pressure, or histopathology were observed between groups. These findings indicate that C. urucurana exerts lipid-lowering, renal, and cardioprotective effects against oxidative stress in a preclinical model of multiple risk factors for heart disease. Laboratory of Preclinical Research of Natural Products Post-Graduate Program in Medicinal Plants and Phytotherapeutics in Basic Attention Paranaense University Laboratory of Preclinical Research of Natural Products Post-Graduate Program in Animal Science with Emphasis on Bioactive Products Paranaense University Laboratory of Cardiovascular Pharmacology Faculty of Health Sciences Federal University of Grande Dourados Institute of Research Pelé Pequeno Príncipe Pequeno Príncipe Faculty Laboratory of Histology Faculty of Health Sciences Federal University of Grande Dourados Laboratory of Genetics and Cell Therapy São Paulo State University Laboratory of Preclinical Research of Natural Products Post-Graduate Program in Medicinal Plants and Phytotherapeutics in Basic Attention Post-Graduate Program in Animal Science with Emphasis on Bioactive Products Laboratory of Genetics and Cell Therapy São Paulo State University

Ethnopharmacological relevance Cecropia pachystachya Trecul (Urticaceae) is a medicinal plant popularly known as ‘embauba’. In Brazil, the leaves of this species are used for the treatment of various kidney and cardiovascular diseases. However, there are no detailed studies on the renal and cardiovascular activities of this species. No studies on the anatomy or the quality control of this herbal drug is available thus far. Aim This study was aimed to investigate the ethnopharmacological properties of the leaves of C. pachystachya. Material and methods The leaves of C. pachystachya were analyzed by light and scanning electron microscopy for pharmacobotanical and anatomical characterization. The ethanol-soluble fraction of C. pachystachya leaf extract (ESCP) was characterized by high-performance liquid chromatograph equipped with diode array detector and mass spectrometry (HPLC-DAD-MS). The acute oral toxicity of ESCP on female Wistar rats was assessed. The acute and prolonged diuresis and antioxidant effects of ESCP (30, 100, and 300 mg/kg) were evaluated in male Wistar rats. In addition, the hypotensive effects of the ESCP as well as the vasodilatory activity in isolated and perfused mesenteric vascular beds were investigated. Results The anatomical markers obtained in this study can help in the identification of C. pachystachya, as well as to distinguish it from the other ‘embaubas’. The metabolites found in the ESCP were phenolic compounds, mainly C- and O-glycosylated flavonoids. The ESCP did not exhibit any toxic effects at a dose of 2000 mg/kg. Significant diuretic activities were observed at the doses of 30, 100, and 300 mg/kg. In addition, a significant modulating activity of the tissue redox state was observed after prolonged treatment. On the other hand, no hypotensive or vasodilator activity was observed. Conclusion The key findings of the present study can contribute to the taxonomy, species identification and quality control of C. pachystachya. Chemical studies have shown the presence of glycosylated flavonoids, phenylpropanoid derivative and proanthocyanidins. The pharmacological studies showed significant diuretic and antioxidant effects of C. pachystachya leaf extract, indicating a possible validation of its popular medicinal use.

Ethnopharmacological relevance Aloysia polystachya (Griseb) Moldenke (Verbenaceae), popularly known as “burrito”, is a South American species widely prescribed by local Brazilian healers for the treatment of cardiovascular diseases. However, its antihypertensive and cardioprotective effects are still unknown. Aim To evaluate the role of the ethanol-soluble fraction of A. polystachya leaves (ESAP) against hypertension in spontaneously hypertensive rats (SHRs), as well as its safety, morphoanatomical and phytochemical aspects. Materials and methods First, the leaves and stems of A. polystachya were analyzed by optical and scanning electron microscopy in order to provide anatomical data for quality control. Then, ESAP was obtained and its chemical profile was analyzed by LC-DAD-MS. In addition, the cytotoxic and acute toxicity potential of ESAP were evaluated in six cell lines and in female Wistar rats, respectively. Next, female spontaneously hypertensive rats (SHRs) received ESAP (30, 100, 300 mg/kg), hydrochlorothiazide (25 mg/kg), or vehicle once daily for 28 days. Weekly kidney function was monitored by analyzing urinary parameters. At the end of the 28-day treatment, the electrocardiographic profile, blood pressure, and renal and mesenteric vascular reactivity were evaluated. Relative organ (heart, kidney, and liver) weights and biochemical parameters were also evaluated. Finally, the heart, kidneys, and aorta were collected for determination of the tissue redox state, cardiac morphometry, and histopathological analysis. Results The chemical profile of ESAP was composed by organic acids, a nucleoside, methoxylated flavones and glycosylated compounds including phenolic acids, phenylpropanoids, iridoids and monoterpenes. No signs of toxicity were observed in all cell's lines nor in female Wistar rats submitted to this trial. All SHRs from the negative control group presented a reduction in renal function, alterations in the renal and mesenteric vascular reactivity, and electrocardiographic and morphometric changes typical of ventricular hypertrophy. Oral prolonged ESAP-administration in SHRs was able to reverse renal, electrocardiographic and hemodynamic changes induced by hypertension. Moreover, ESAP-treatment was able to modulate the vascular and renal arterial reactivity and tissue redox state. The aforementioned data were accompanied by reduction of cardiac hypertrophy. Conclusion In this study, we present important anatomical and phytochemical data that contributed to the correct identification and quality control of A. polystachya. In addition, we have shown that ESAP is safe after acute administration and present significant cardioprotective effects (at 30, 100, and 300 mg/kg doses) in SHRs after prolonged treatment.

Ethnopharmacological relevance Although Luehea divaricata Mart. (Malvaceae) is popularly used by the population of the Brazilian Pantanal for the treatment of different types of kidney diseases, no study has been carried out to prove this ethnobotanical indication. Aim To investigate the possible cardiorenal effects of an herbal preparation obtained from L. divaricata leaves. Materials and methods First, to provide quality control standards, a detailed morphological and microchemical characterization of L. divaricata leaves was performed. Then, the purified aqueous extract was obtained from the leaves of this species (ESLD) and a thorough phytochemical characterization was performed. Subsequently, acute oral toxicity test was performed after single administration of different doses (5, 50, 300, 2000 mg/kg) in male and female Wistar rats. Finally, the diuretic, hypotensive and antioxidant properties of ESLD (30, 100, 300 mg/kg) were evaluated after acute and prolonged treatment and the role of angiotensin converting enzyme, aldosterone, vasopressin, and nitric oxide in these effects was investigated. Results Analyses performed by liquid chromatography–mass spectrometry showed that the main secondary metabolites present in ESLD were flavonol O-glycosides and flavone C-glycosides. Acute and prolonged treatment with ESLD was able to expressively increase urinary volume and electrolyte excretion. Mean blood pressure and systolic blood pressure were also significantly reduced after acute treatment. Moreover, treatment with ESLD was able to reduce thiobarbituric acid reactive species and increase serum nitrate levels. Conclusion The data obtained showed that ESLD has an important diuretic and hypotensive effect, which is probably dependent on the reduction of oxidative stress and increased bioavailability of nitric oxide.

Bupivacaine is the most employed local anesthetic in surgical procedures, worldwide. Its systemic toxicity has directed the synthesis of the less toxic, S(−) enantiomer. This work describes a formulation of ionic gradient liposomes (IGL) containing S75BVC, an enantiomeric excess mixture of 75% S(−) and 25% R(+) bupivacaine. IGL prepared with 250 mM (NH4)2SO4 in the inner aqueous core of phosphatidylcholine and cholesterol (3:2 mol%) vesicles plus 0.5% S75BVC showed average sizes of 312.5 ± 4.5 nm, low polydispersity (PDI

Lipid nanocapsules (LNC) are special drug delivery system (DDS) carriers obtained by the phase-inversion temperature method (PIT). This study describes the encapsulation of the local anesthetics (LA) prilocaine (PLC) and lidocaine (LDC) in lipid nanocapsules (LNCPLC+LDC) optimized by 23 factorial design, characterized through DLS, NTA, CRYO-EM and release kinetics and incorporated in carbopol gel (GelLNC PLC+LDC) prior to in vivo anesthetic effect (in mice) evaluation. A very homogeneous population of small (50 nm; polydispersity index = 0.05) spherical nanocapsules with negative zeta potentials (−21 mV) and ca. 2.3 × 1015 particles/mL was obtained. The encapsulation efficiency was high (81% and 89% for prilocaine and lidocaine, respectively). The release rate profile was free PLC = free LDC > LNCPLC+LDC > GelLNC PLC+LDC. The hybrid system increased (4x) the anesthesia time in comparison to an equipotent gel formulation prepared without LNC. No tissue damage was detected on the tail skin of mice that received the formulations. This study shows that lipid nanocapsules are suitable carriers for PLC and LDC, promoting longer and safer topical anesthesia. GelLNC PLC+LDC is mucoadhesive and suitable for application in the mouth, where it could be used as a pre-anesthetic, to reduce pain of needle stick (infiltrative anesthesia).

Although Acanthospermum hispidum is used in Brazilian folk medicine as an antihypertensive, no study evaluated its effects on a renovascular hypertension and ovariectomy model. So, this study investigated the mechanisms involved in the antihypertensive effects of an ethanol-soluble fraction obtained from A. hispidum (ESAH) using two-kidney-one-clip hypertension in ovariectomized rats (2K1C plus OVT). ESAH was orally administered at doses of 30, 100, and 300 mg/kg, daily, for 28 days, after 5 weeks of surgery. Enalapril (15 mg/kg) and hydrochlorothiazide (25 mg/kg) were used as standard drugs. Diuretic activity was evaluated on days 1, 7, 14, 21, and 28. Systolic, diastolic, and mean blood pressure and heart rate were recorded. Serum creatinine, urea, thiobarbituric acid reactive substances, nitrosamine, nitrite, aldosterone, vasopressin levels, and ACE activity were measured. The vascular reactivity and the role of nitric oxide (NO) and prostaglandins (PG) in the vasodilator response of ESAH on the mesenteric vascular bed (MVB) were also investigated. ESAH treatment induced an important saluretic and antihypertensive response, therefore recovering vascular reactivity in 2K1C plus OVT-rats. This effect was associated with a reduction of oxidative and nitrosative stress with a possible increase in the NO bioavailability. Additionally, a NO and PG-dependent vasodilator effect was observed on the MEV.

Therapeutic approaches for the treatment of dyslipidemia and atherosclerosis have radically changed in recent decades. Part of this advance undeniably stems from basic biomedical research that has provided a better understanding and identification of new therapeutic targets. The aim of this work was to develop a model to induce atherogenesis and hepato-renal impairment in female Wistar rats. The following groups received the respective treatments for 60 days: control animals, non-ovariectomized rats that received an atherogenic diet (NEAD), ovariectomized rats that received an atherogenic diet (NOAD), non-ovariectomized rats that received an atherogenic diet and oral N&omega, nitro-l-arginine methyl ester hydrochloride (l-NAME, LEAD), and ovariectomized rats that received an atherogenic diet and oral l-NAME (LOAD). Animals in the NEAD, NOAD, LEAD, and LOAD groups also received methimazole and cholecalciferol daily. Urinary, biochemical, hemodynamic, and electrocardiographic parameters and renal function were assessed. Samples of the liver, heart, kidney, and arteries were collected to investigate redox status and perform histopathological analyses. All of the groups developed dyslipidemia and hepatic steatosis. Only the NEAD group developed arterial lesions that were compatible with fatty streaks. Renal function was significantly impaired in the LEAD and NOAD groups. These results indicate a viable alternative to induce atherogenesis and hepato-renal impairment in female rats.

Ethnopharmacological relevance Plinia cauliflora (Mart.) Kausel (Myrtaceae) is popularly known as “jaboticaba” or “jaboticaba”. The fruit is appreciated for both fresh consumption and the manufacture of jelly, juice, ice cream, fermented beverages, and liqueurs. The more widespread traditional use of the plant involves the treatment of diarrhea, which utilizes all parts of the plant, including the fruit peels. Aim of the study: We sought to elucidate possible risks of the administration of an ethanol-soluble fraction that was obtained from an infusion of P. cauliflora fruit peels (SEIPC). We performed a series of experiments to evaluate possible toxicity, in which we administered SEIPC orally both acutely and repeatedly for 28 days. We also evaluated possible endocrine-disruptive and genotoxic effects in eukaryotic cells. The possible mutagenic activity of SEIPC was evaluated using reverse mutation (Ames) assays. Materials and methods SEIPC was produced and chemically characterized by LC-DAD-MS. Acute toxicity and behavioral and physiological alterations were evaluated in the modified Irwin test. Respiratory rate, arterial blood gas, electrocardiography, respiratory rate, heart rate, and blood pressure were evaluated, and hematological, biochemical, and histopathological analyses were performed after 28 days of oral treatment. The comet assay, mammalian erythrocyte micronucleus test, uterotrophic test, Hershberger bioassay, and AMES test were performed using appropriate protocols. Results From SEIPC, ellagic acid and derivatives, flavonols and anthocyanidins, as well as citric acid and gallic acid, were annotated by LC-DAD-MS. We did not observed any significant toxic effects after acute or prolonged SEIPC treatment. No endocrine-disruptive or mutagenic effects were observed. Conclusions The present study found that SEIPC did not cause any significant alterations of various corporeal systems, including cardiac electrical activity, body temperature, respiratory rate, and arterial pressure. No alterations of biochemical, hematological, or blood gas parameters were observed. SEIPC did not cause any perturbations of the endocrine system or mutagenic, cytotoxic, or genotoxic effects. These findings substantiate the safe clinical use of P. cauliflora.

Talisia esculenta (A. St.-Hil.) Radlk. is a large tree belonging to family Sapindaceae and popularly known as “pitombeira” or “pitomba.” Although species have relevant economic and medicin...

Ethnopharmacological relevance Talinum paniculatum (Jacq.) Gaertn. (Talinaceae), popularly known as “major gomes” and “erva gorda”, is a non-conventional food plant extensively distributed throughout the Brazilian territory. In Brazilian folk medicine, this species is used as aphrodisiac, to treat gastrointestinal problems, and as a cardioprotective agent. However, there are no reports in the literature proving its cardiovascular effects. Aim To perform a whole-ethnopharmacological investigation of the cardiorenal properties of the ethanol soluble fraction from T. paniculatum (ESTP) in Wistar rats. Material and methods First, plant samples were collected, properly identified and a morpho-anatomical characterization was carried out to provide quality control parameters. Then, ESTP was obtained and its chemical profile was determined by LC-DAD-MS. In addition, an acute toxicity assay was conducted in female Wistar rats in order to observe any toxic effects after one single administration. Finally, the diuretic and hypotensive potential of ESTP (30, 100 and 300 mg/kg) were investigated in male rats followed by the evaluation of its possible effects on peripheral vascular resistance. Results Chemical compounds identified from ESTP were chlorogenic acids, amino acids, nucleosides, O-glycosylated flavones and organic acids. No signs of toxicity as well as no changes in urine volume or electrolyte elimination were observed after ESTP acute treatment. On the other hand, prolonged treatment with all doses of ESTP significantly increased urine volume and electrolyte excretion (Na+, K+ and Cl−) without affecting blood pressure or heart rate. Apparently, these effects are involved with the activation of the small conductance calcium-activated potassium channels contributing to the increase of renal blood flow and glomerular filtration rate. Conclusion Data presented show important information about the ethnomedicinal properties of T. paniculatum. In addition, the study presents the ESTP as a possible herbal medicine, especially when a sustained diuretic effect is required.

Campomanesia pubescens is a fruit plant widely distributed in South America and used by the population for medicinal and nutritional purposes, with important economic and cultural value. This study evaluated the toxic potential of the ethanolic extract from C. pubescens (EEFCP) fruits through acute and short-term toxicity tests. For the acute toxicity test, female rats received a single oral dose of 2000 mg/kg body weight of EEFCP and were observed for 14 days. In the short-term toxicity test, male and female rats received repeated oral doses of 125, 250, 500 or 1000 mg/kg of EEFCP, being treated and observed for 28 days, and after the treatment period, a satellite and satellite control group remained under observation for another 14 days. No mortality, clinical and organ weight alterations were observed, indicating that LD50 is greater than 2000 mg/kg body weight. In addition, the doses tested did not produce significant changes in the behavioral, physiological, hematological or histopathological parameters of animals. These results demonstrate the low acute and short-term toxicity of EEFCP in rats. The data obtained are of great relevance since they provide important information about a plant species of great economic, nutritional and ethnopharmacological value.

Ethnopharmacological relevance Gomphrena celosioides Mart., belonging to the Amaranthaceae family, is a weed known as “perpetua,” and its ethnopharmacological use is to treat of urinary tract disorders and kidney stones. Urinary tract disorders and kidney stones could include several pathological conditions such hypertension, diuretic and lithiasic problems. In the present work a model of renovascular hypertension was developed in vivo to investigate its usefulness as an antihypertensive drug. Aim of the study Evaluate the effect of acute and 28 day oral administration of G. celosioides extract on systemic arterial pressure and diuresis of renovascular-hypertensive rats, as well as its effect on cardiac remodeling and vascular reactivity. Materials and methods Ethanolic extract of G. celosioides (EEGC) was used. To induce renovascular hypertension, adult male Wistar rats were submitted to Goldblatt 1K1C or 2K1C surgery. The mean arterial pressure (MAP) of 1K1C animals was directly assessed by cannulation of the carotid artery before and after intraduodenal acute administration of 30, 100 or 300 mg/kg of EEGC. For the 4-week assay, 2K1C animals received daily treatments with water (control group), 100 mg/kg EEGC or 15 mg/kg enalapril for 28 days. Diuresis and caudal blood pressure were assessed weekly, and at the 28th day of treatment, the MAP was directly quantified shortly before euthanasia. Internal organs were removed, weighed and routinely processed for histology and the left ventricle wall was measured. Blood was collected for biochemical analysis and mechanism investigation by quantification of angiotensin converting enzyme (ACE) activity and aldosterone, nitrite and thiobarbituric acid reactive substances (TBARS) concentration. The rats’ mesenteric beds were isolated and cannulated to have their pressure variation assessed after crescent doses of phenylephrine (Phe), acetylcholine (ACh) and sodium nitroprusside (SNP). Results EEGC acutely reduced MAP the dose of 100 mg/kg. In the 4-week assay, EEGC acted as diuretic after acute administration after 1, 2, 3 and 4 weeks of treatment. EEGC also acted as an antihypertensive and it showed significant difference already after 1 week (and after 3 and 4 weeks) compared to control, with its MAP close to pre-surgery values at the end of the experiment. It promoted ACE inhibition, which led to lower aldosterone levels. The lower TBARS and higher nitrite concentration found in the EEGC group suggest antioxidant activity and NO maintenance. Moreover, EEGC counteracted the impairment of vascular reactivity induced by renovascular hypertension. The extract group presented thinner left ventricle wall compared to the control, meaning reduced hypertension-induced cardiac remodeling. Conclusions The G. celosioides diuretic effect is maintained on renovascular hypertensive rats and can reduce the blood pressure after the first week of treatment by inhibiting ACE and these effects are longstanding and strong enough to promote protection against cardiac remodeling. Therefore, it shows potential as an antihypertensive drug.

Casearia sylvestris Sw., Salicaceae, is an important medicinal plant widely used in Brazil for the treatment of various cardiovascular disorders. This species was included as of interest by Brazilian Unified Health System. Although preclinical studies described cardiovascular protective effects and apparent absence of toxicity, no studies have evaluated its carcinogenic potential. In this study, we proposed a short-term carcinogenesis evaluation of C. sylvestris in Wistar rats, aiming to check the safety of this species to use it as proposed by Brazilian Unified Health System. C. sylvestris leaves were obtained and the crude extract was prepared by maceration from methanol/water. Wistar rats were orally treated for 12 weeks with 50, 250 or 500 mg kg−1 of crude extract or vehicle. Body weight, daily morbidity and mortality were monitored. Blood and bone marrow samples were collect for micronucleus test, comet assay and tumor markers evaluation. Vital organs were removed to macro and histopathological analyses. The crude extract did not induce mutagenic and genotoxic effects and no alterations were observed in important tumor markers. Finally, no detectable signs of injury through gross pathology or histopathological examinations were observed. Our results certify the absence of the crude extract toxicity, indicating its safety, even at prolonged exposure as proposed by Brazilian Unified Health System. Keywords: Genotoxicity, Mutagenesis, Short-term carcinogenesis, Toxicity, Tumor markers

The objective of this study was to evaluate the maternal, embryotoxic and teratogenic effects of Caryocar brasiliense pulp oil (OPCB), oil widely used in Brazilian cuisine and traditional medicine. Pregnant Wistar female rats were used in this study for three treatment groups (250, 500 and 1000 mg/kg/day) and a control group. The OPCB was administered orally throughout the period of organogenesis of females (6th until the 15th day of gestation). The pregnant females were gross necropsied on d20, followed by maternal and fetus examination, to evaluate the teratogenicity, reproductive and developmental performance of OPCB. The results showed there was no significant statistical difference in the ponderal evolution of the pregnant females, as well as in the behavioral, hematological, biochemical or histopathological data, indicating the absence of maternal toxicity of the oil. The mean number of corpora lutea, implantation and resorption sites, as well as all calculated reproductive rates, also remained statistically similar between the groups, indicating low embryotoxic effects of the tested plant specie. In fetal examination, external anomalies and skeletal abnormalities were observed in all treated and control groups. The NOAEL for maternal toxicity and embryo/fetal development for the OPCB administered by gavage, was 1000 mg/kg/bw/day.

Ethnopharmacological relevance Acanthospermum hispidum DC. is an important medicinal herb that belongs to family Asteraceae, popularly used as a diuretic and hypotensive in the region of Pantanal, state of Mato Grosso do Sul, Brazil. Despite the relevance of this species throughout the country, there are no detailed studies about its possible ethnobotanical indication. Aim To carry out a detailed ethnopharmacological investigation of the cardio-renal properties of A. hispidum. Materials and methods First, a detailed morpho-anatomical study with the purpose of characterizing and providing quality control parameters for the species was carried out. Then, purified aqueous extract (ESAH) was obtained and a detailed phytochemical investigation about its main secondary metabolites was performed. In addition, a thorough acute toxicological study was conducted to evaluate the actual toxic effects of this preparation. Finally, the possible diuretic and hypotensive effects of ESAH on male Wistar rats (30, 100, 300 mg/kg; intraduodenally) were evaluated, and using pharmacological antagonists or inhibitors, the involvement of prostaglandin/cAMP and nitric oxide/cGMP pathway and potassium channels in ESAH-induced hypotension was investigated. Results The analyses performed by liquid chromatography–mass spectrometry showed that the main secondary metabolites present in ESAH were phenolic compounds, such as caffeoylquinic acids (chlorogenic acid), dicaffeoylquinic acids and glycosylated flavonoids (quercetin glucoside and galactoside). ESAH did not induce any acute toxic effects and did not affect the urinary volume or renal excretion of electrolytes in Wistar rats. On the other hand, intraduodenal administration of ESAH induces a significant acute hypotensive effect. Previous treatment with N(G)-nitro- L -arginine methyl ester, methylene blue, or tetraethylammonium fully avoided the hypotensive effect of ESAH. All other parameters were not affected by treatment with ESAH. Conclusion Data obtained in this study allow us to suggest that ESAH obtained from A. hispidum presents an important acute hypotensive effect, which appears to be dependent on the nitric oxide/cGMP pathway. This study presents new evidences about the therapeutic potential of this species when acute hypotensive response is required.

Attalea phalerata Mart. ex Spreng., popularly known as “bacuri”, is a native plant from the brazilian Cerrado and used in folk medicine as a pulmonary decongestant, an anti-inflammatory for joints and antipyretic. There is an expectation about the use in chronic disease of the Attalea phalerata oil since its composition is high in carotenoids and beneficial fatty acids. The aim of the study was to evaluate the toxicological profile of the oil extracted from Attalea phalerata Mart. ex Spreng. pulp (APO). Acute and subacute toxicity studies were performed in male and female Wistar rats according to the OECD - Guidelines 425 and 407. For the acute toxicity, one single dose of the APO (2000 mg/kg) was administered by gavage to five female rats. In the subacute toxicity, four different doses (125, 250, 500 and 1000 mg/kg) of the APO were administered to male and female rats for 28 consecutive days. No deaths or behavioral changes were observed during both experiments as well as no changes in organ weights, hematological, histopathological parameters. The biochemical parameters showed changes in phosphatase alkaline and albumin levels, however these values are within the normal range for the species. A significant reduction in cholesterol and triglycerides was also observed in some of the animals treated with the APO. Therefore, the LD50 is higher than 2000 mg/kg and the APO oil can be considered safe at the doses tested in rats. However, further assessments are required in order to proceed to clinical studies in humans.

The Caryocar brasiliense (pequi) is a Brazilian fruit of important geographic distribution and of broad popular use for nutritional purposes. This study aimed to evaluate the toxicological potential of pequi through the acute and subchronic toxicity tests. For the acute toxicity test, female Wistar rats received, orally, a single dose of 2000 mg/kg/bw of pequi oil and were observed for 14 days. In subchronic toxicity test, Wistar male and female rats received, orally, repeated doses of 125, 250, 500 or 1000 mg/kg/bw of the oil, being treated and observed for 28 days. In the acute toxicity test, no changes as well as no mortality were observed, indicating that the LD50 is higher than 2000 mg/kg/bw. In the subchronic toxicity test, the tested doses produced no significant changes in behavioral, physiological, biochemical or histopathologic parameters in animals. Some hematological abnormalities were found after subchronic exposure and need to be clarified. These results demonstrate the low toxicity of acute and subchronic to the oil of pequi in rats. However, additional studies are required in order to verify if the hematological abnormalities have any relation to the oil exposure and also provide sufficient safety evidence for the use of the oil in humans.

Ethnopharmacological relevance Alibertia edulis, popularly known as “marmelo do Cerrado” is a native plant from the brazilian Cerrado. It has high food and ornamental potential and the tea leaves are currently used as hypoglycemic, antihypertensive and diuretic. Aim of the study In order to evaluate the safety of the aqueous extract of Alibertia edulis leaves (AEAE), the acute and subacute toxicity tests were performed in male and female Wistar albino rats. Materials and methods The experiments were performed in accordance with the OECD guidelines 425 and 407. For the acute toxicity, one single dose of the AEAE (2000 mg/kg) was administered by gavage to five female rats. The animals were observed for 14 days for any signs of toxicity and death. In the subacute toxicity, four different doses (125, 250, 500 and 1000 mg/kg) of the AEAE were administered to male and female rats for 28 consecutive days. A satellite group received the maximum dose (1000 mg/kg) for 28 days and remained untreated for 14 more days in order to observe reversibility, persistence, or delayed occurrence of toxic effects. The five parameters of the Hippocratic screening, body weight, food and water intake were daily observed. At the end of the experiment, blood samples were collected for the hematological and biochemical analysis. The vital and reproductive organs were inspected for any histopathological changes. Results No deaths or behavioral changes were observed during both experiments as well as no changes on organ weights, biochemical, hematological and histopathological parameters that could indicate any signs of toxicity. Conclusion All doses tested can be considered safe in rats and the LD50 is higher than 2000 mg/kg. Therefore, further assessments are required in order to proceed to clinical studies in humans.

A retrospective study has been conducted for a period of 13 years, between 2000 and 2012, on files archived at the Laboratorio de Anatomia Patologica (LAP), at the Faculdade de Medicina Veterinaria e Zootecnia (FAMEZ) at Universidade Federal de Mato Grosso do Sul (UFMS). For this study, 2.359 1