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ABSTRACT Introduction Increasing urbanization has particularly affected rivers and their outer edges in cities, including Kathmandu Valley, which encompasses Lalitpur, the nation's third‐largest city. This study aims to conduct a parasitological survey to investigate the occurrence of zoonotic intestinal protozoa and helminths along the Karmanasa River bank in central Nepal. Methods Faecal samples from openly defaecating animals were collected via non‐invasive techniques, and coproscopy was carried out using direct wet mount, concentration and acid‐fast staining methods to ensure reliable findings. Results The findings showed that all the faecal samples were positive for intestinal parasites as follows: buffaloes (15/15), cats (5/5), cattle (30/30), chickens (7/7), dogs (15/15), goats (15/15), pigs (20/20) and rats (12/12). A total of 28 intestinal parasites were reported, out of which 21 species possess zoonotic potentialities, and each host was reported to harbour at least one zoonotic parasite. Entamoeba sp., Cryptosporidium sp., hookworm, Trichuris sp., Trichostrongylus and Balantidium coli were among the predominant zoonotic parasites. The use of the landscape for livestock grazing and the presence of free‐ranging animals could have all added to the zoonotic risks. Moreover, the excretion of a moderate to large number of zoonotic parasite eggs in the faecal samples indicates a transmission risk. Conclusions The study detected 21 species of intestinal zoonotic parasites circulating along the landscape in the study area, indicating a higher risk of cross‐transmission. Therefore, strategic treatment of livestock and free‐ranging animals and periodic public health awareness programs for the local inhabitants are highly recommended.

ABSTRACT Background The placement of ligatures in the cervical area of rat molars is considered as a predictable model to induce periodontitis. Objectives The present explorative study aimed to compare the efficacy of metal wires (MWs), without or with sandblasting, versus silk ligatures (SLs) in inducing periodontal bone loss in rats. Materials and Methods Twenty‐four Wistar rats were randomly divided into three groups of eight rats that received three different types of ligatures (MW, sandblasted wire [SMW], and SL) around their first right mandibular molar, while the contralateral tooth was left without the ligature and served as a control. Bone loss was assessed by measuring the distance from the cementoenamel junction (CEJ) to the bone crest at the distal aspect of the first molar on central mesiodistal sections generated from micro‐CT scans taken 24 and 35 days after ligature placement. Results In the SL group, only in two rats the ligatures were retained until the end of the 24‐day period; in all other animals, the ligatures were lost at some time point. In the SMW, the ligatures were retained only for the 24‐day period. In the MW group, no ligatures were lost. Irrespective of the group or experimental period, the difference in the crestal bone level between ligated and control teeth was in most cases z

Abstract Current rodent models of anorexia nervosa (AN) often have accelerated weight loss that often do not allow for investigation of physiological ramifications of prolonged low weight status characteristic of AN. The purpose of this project was to refine a rodent model of AN to extend the duration of low weight status and allow for investigation of recovery. Eight‐week‐old female Sprague Dawley rats underwent 50%–60% food restriction for 30 days. Rats were group‐housed except during feeding, where AN rats were individually housed and given up to 2 h to consume food. Control (CON) rats were allowed to consume food ad libitum. To simulate recovery, a separate cohort of animals underwent the same food restriction protocol for 30 days, then rats (AN‐R) were allowed to consume food ad libitum to facilitate weight recovery for an additional 30 days. AN‐R rats were compared to age matched controls (CON‐R). AN rats lost ~15% bodyweight and were ~30% lighter than CON. Compared to CON, AN rats had ~35% lower fat content, ~18% lower bone mineral density, ~22% smaller plantaris muscle mass and ~52% smaller ovaries. Upon reintroduction of food, AN‐R rats achieved comparable bodyweights to CON‐R rats after ~10 days. However, after 30 days of recovery, AN‐R rats still had ~14% lower bone mineral density and ~11% smaller plantaris mass and ~21% smaller ovaries. This refinement of rodent AN results in physiological side effects of AN without reaching excessive weight loss requiring euthanasia. Moreover, some physiological consequences of simulated AN are not resolved with weight restoration.

Abstract Objectives Frankincense (Luban) is a resin obtained from trees of genus Boswellia. The south of Oman hosts Boswellia sacra trees known to have many social, religious and medicinal uses. The anti‐inflammatory and therapeutic potential of Luban has recently attracted the interest of the scientific community. The aim is to study the efficacy of Luban water extract and its essential oils on experimentally induced renal stones in rats. Materials and Methods A rat model of urolithiasis induced by trans‐4‐hydroxy‐L‐proline (HLP) was used. Wistar Kyoto rats (27 males, 27 females) were randomly distributed into nine equal groups. Treatment groups were given Uralyt‐U (standard) or Luban (50, 100 and 150 mg/kg/day), starting Day 15 from HLP induction for a duration of 14 days. The prevention groups were given Luban in similar doses, starting Day 1 of HLP induction for 28 days. Several plasma biochemical and histological parameters were recorded. Data were analysed with GraphPad Software. Comparisons were performed by one‐way analysis of variance (ANOVA) and the Bonferroni test. Results The lithogenic effects of HLP, such as an increase in urine oxalate and cystine, an increase in plasma uric acid and an increase in kidney levels of calcium and oxalate, have all been best significantly reversed by the Luban dose of 150 mg/kg/day. The histological changes of HLP on the kidney tissue including calcium oxalate crystal formation, cystic dilatation, high degree of tubular necrosis, inflammatory changes, atrophy and fibrosis have also been ameliorated by Luban dose of 150 mg/kg/day. Conclusion Luban has shown a significant improvement in the treatment and prevention of experimentally induced renal stones, particularly at a dose of 150 mg/kg/day. Further studies on the effect of Luban in other animal models and humans with urolithiasis are warranted.

Abstract Background The upper limit for liver resections in rats is approximately 90%. In the early postoperative phase, mortality increases. The aim of the present study was to validate the rat model of 90% partial hepatectomy (PH) as a model of post‐hepatectomy liver failure (PHLF). Further, we wanted to test a quantitative scoring system as a detector of lethal outcomes caused by PHLF in rats. Methods Sixty‐eight rats were randomized to 90% PH, sham operation, or no surgery. Further, block randomization was performed based on time of euthanization: 12, 24, or 48 h after surgery. A general distress score (GDS) ≥10 during the day or ≥6 at midnight prompted early euthanization and classification as nonsurvivor. Animals euthanized as planned were classified as survivors. During euthanization, blood and liver tissue were collected, and liver‐specific biochemistry was evaluated. Results Based on the biochemical results, all animals subjected to 90% PH experienced PHLF. Seventeen rats were euthanized due to irreversible PHLF. The GDS increased for nonsurvivors within 12–18 h after surgery. The mean time for euthanization was 27 h after surgery. Conclusion Based on the GDS and liver‐specific biochemistry, we concluded that the model of 90% PH seems to be a proper model for investigating PHLF in rats. As a high GDS is associated with increased mortality, the GDS appears to be valuable in detecting lethal outcomes caused by PHLF in rats.

Abstract Heart rate variability (HRV) is commonly used in experimental studies to assess sympathetic and parasympathetic activities. The belief that HRV in rodents reflects similar cardiovascular regulations in humans is supported by evidence, and HRV in rats appears to be at least analogous to that in humans, although the degree of influence of the parasympathetic division of the autonomic nervous system (ANS) may be greater in rats than in humans. Experimental studies are based on control or baseline values, on the basis of which the change in ANS activity after a given experimental intervention is assessed, but it is known that the ANS in rats is very sensitive to various stress interventions, such as the manipulation itself, and ANS activity can also differ depending on sex, the time of measurement, and whether the animals are under general anaesthesia. Thus, for correct assessment, changes in ANS activity and their relationship to the observed parameter should be based on whether ANS activity does or does not change but also to what extent the activity is already changed at the start of the experiment. Since rats are considered to be the most suitable model animal for basic cardiovascular research, in this review we point out existing differences in individual HRV frequency parameters at the start of experiments (control, baseline values), taking into account sex in relation to time of measurement and anaesthesia.

Abstract While phosphodiesterase‐5 inhibition (PED5i) may prevent hypertrophy and failure in pressure‐overloaded heart in an experimental model, the impact of PDE5i on volume‐overload (VO)‐induced hypertrophy is unknown. It is also unclear whether the hypertrophied right ventricle (RV) and left ventricle (LV) differ in their responsiveness to long‐term PDE5i and if this therapy affects renal function. The goal of this study was to elucidate the effect of PDE5i treatment in VO due to aorto‐caval fistula (ACF) and to compare PDE5i treatment with standard heart failure (HF) therapy with angiotensin‐converting enzyme inhibitor (ACEi). ACF/sham procedure was performed on male HanSD rats aged 8 weeks. ACF animals were randomized for PDE5i sildenafil, ACEi trandolapril, or placebo treatments. After 20 weeks, RV and LV function (echocardiography, pressure‐volume analysis), myocardial gene expression, and renal function were studied. Separate rat cohorts served for survival analysis. ACF led to biventricular eccentric hypertrophy (LV: +68%, RV: +145%), increased stroke work (LV: 3.6‐fold, RV: 6.7‐fold), and reduced load‐independent systolic function (PRSW, LV: −54%, RV: −51%). Both ACF ventricles exhibited upregulation of the genes of myocardial stress and glucose metabolism. ACEi but not PDE5i attenuated pulmonary congestion, LV remodeling, albuminuria, and improved survival (median survival in ACF/ACEi was 41 weeks vs. 35 weeks in ACF/placebo, p = .02). PDE5i increased cyclic guanosine monophosphate levels in the lungs, but not in the RV, LV, or kidney. PDE5i did not improve survival rate and cardiac and renal function in ACF rats, in contrast to ACEi. VO‐induced HF is not responsive to PDE5i therapy.

Abstract Vibration acceleration (VA) using a whole‐body vibration device is beneficial for skeletal muscles. However, its effect at the cellular level remains unclear. We aimed to investigate the effects of VA on muscles in vitro and in vivo using the C2C12 mouse myoblast cell line and cardiotoxin‐induced injury in male rat soleus muscles. Cell proliferation was evaluated using the WST/CCK‐8 assay and proportion of Ki‐67 positive cells. Cell migration was assessed using wound‐healing assay. Cell differentiation was examined by the maturation index in immunostained cultured myotubes and real‐time polymerase chain reaction. Regeneration of soleus muscle in rats was assessed by recruitment of satellite cells, cross‐sectional area of regenerated muscle fibers, number of centrally nucleated fibers, and conversion of regenerated muscle from fast‐ to slow‐twitch. VA at 30 Hz with low amplitude for 10 min promoted C2C12 cell proliferation, migration, and myotube maturation, without promoting expression of genes related to differentiation. VA significantly increased Pax7‐stained satellite cells and centrally nucleated fibers in injured soleus muscles on Day 7 and promoted conversion of fast‐ to slow‐twitch muscle fibers with an increase in the mean cross‐sectional area of regenerated muscle fibers on Day 14. VA enhanced the proliferation, migration, and maturation of C2C12 myoblasts and regeneration of injured rat muscles.

Abstract With the technological advances made to expand space exploration, astronauts will spend extended amounts of time in space before returning to Earth. This situation of unloading and reloading influences human physiology, and readaptation to full weight‐bearing may significantly impact astronauts' health. On Earth, similar situations can be observed in patients who are bedridden or suffer from sport‐related injuries. However, our knowledge of male physiology far exceeds our knowledge of female's, which creates an important gap that needs to be addressed to understand the sex‐based differences regarding musculoskeletal adaptation to unloading and reloading, necessary to preserve health of both sexes. Using a ground‐based model of total unloading for 14 days and reloading at full weight‐bearing for 7 days rats, we aimed to compare the musculoskeletal adaptations between males and females. Our results reveal the existence of significant differences. Indeed, males experienced bone loss both during the unloading and the reloading period while females did not. During simulated microgravity, males and females showed comparable muscle deconditioning with a significant decline in rear paw grip strength. However, after 7 days of recovery, muscle strength improved. Additionally, sex‐based differences in myofiber size existing at baseline are significantly reduced or eliminated following unloading and recovery.

Abstract Caffeine is the most widely consumed psychostimulant drug which could affect learning and memory acting through central adenosine receptors. Although caffeine has been suggested to impair the acquisition and the expression of auditory fear conditioning, its effect on the extinction has not been elucidated. To address this issue, in the present study, we investigated whether caffeine affects the extinction of conditioned fear in an auditory fear conditioning paradigm. During conditioning, rats received pairings of auditory cues (conditioned stimulus, CS) and foot shocks (unconditioned stimulus). On the next day, the rats were intraperitoneally administrated saline or caffeine (5 or 10 mg/kg) and then subjected to the extinction training, in which CSs were repeatedly presented without the foot shocks. Twenty‐four hours later, the rats were re‐exposed to the presentations of CSs (retrieval test). We found an acute caffeine administration dose‐dependently decreased freezing rates during the presentations of CS in the extinction training. Furthermore, caffeine‐treated animals showed lower conditioned freezing responses in the retrieval test. These findings suggest that caffeine facilitates the extinction of conditioned fear.

Abstract Rationale Systemic dehydration negatively alters the expression of vocal fold inflammatory and cell junction markers. These biological changes can have downstream effects on the healing processes of injured vocal folds. In the dermis, reduced hydration prolongs inflammation and delays healing. It is unknown whether this biological effect is observed in vocal fold tissue. Objective To investigate the effects of systemic dehydration on vocal fold healing outcomes following acute, bilateral vocal fold injury in a rodent model. Methods Eighteen systemic dehydrated and 18 euhydrated adult male Sprague Dawley rats experienced bilateral vocal fold injuries or no injury (N = 9/group). Vocal fold gene expression levels of inflammatory mediators and epithelial cell junction markers were measured 24 h post‐injury. Results Pro‐inflammatory gene markers (IL‐1β; TNF‐α) were differentially expressed in response to systemic dehydration with vocal fold injury compared to non‐injury. Epithelial cell junction markers (Cadherin‐3, Desmoglein‐1) also exhibited divergent trends following systemic dehydration, but these data were not statistically significant. Conclusions Systemic dehydration may affect cellular vocal fold healing processes within 24 h. These findings lay the groundwork for further investigation of how hydration status can affect vocal fold tissue recovery and influence clinical care.

Abstract Objective Acute‐on‐chronic liver failure (ACLF) causes organ system failures in patients and increases the risk of mortality. One of the main predictors of ACLF development in patients is the severity of systemic inflammation. The purpose of this study was to explore the effects of resolvin D1 (RvD1) on the rat model of ACLF. Methods The ACLF rats were induced by first intraperitoneally (ip) injecting CCl4 and porcine serum for 6 weeks to establish the chronic liver injury, followed by once administration (ip) of lipopolysaccharide and d‐galactose d‐GalN to cause acute liver injury (ALI). An hour before the ALI‐induced treatment, rats were administrated (ip) with 0.9% saline or different doses of RvD1 (0.3 or 1 µg/kg). Afterward, the control and treated rats were killed and samples were collected. Biochemical analysis, hematoxylin‐eosin and Sirius red staining, flow cytometry assay, and real‐time polymerase chain reaction were used to assess the rat liver histopathological injury, the percentage of Treg cells in the spleen, and the messenger RNA (mRNA) levels of transcription factors and immunologic cytokines in liver. Results The necroinflammatory scores and the serum levels of transaminase significantly increased in ACLF rats compared with those in control rats. These impaired changes observed in ACLF rats could be attenuated by the administration of a low dose of RvD1 before the induction of ALI, which was associated with the increased proportion of regulatory T cells (Treg) in the spleen together with the increased gene expression ratio of Foxp3/RORγt and decreased mRNA level of Il‐17a and Il‐6 in the liver. Conclusion A low dose of RvD1 can promote the resolution of inflammation in ACLF rats by increasing the proportion of Treg cells. RvD1, therefore, may be used as a potential drug for the treatment of patients with ACLF.

Abstract Introduction This study aimed to investigate the treatment effect of G protein‐coupled receptor 30 (GPR30) agonist G1 combined with hypothermia (HT) on cognitive impairment and anxiety‐like behavior after subarachnoid hemorrhage (SAH) in rats. Methods Fifty male rats were randomly assigned to one of five groups: Sham group, SAH group, SAH + G1 group, SAH + HT group, and SAH + G1 + HT group. The SAH rat model was established by modified endovascular puncture in all groups except the Sham group. Neurological function after the operation was assessed by Garcia scoring. The degree of rat cerebral edema was determined using dry‐wet weighing method on the 28th day after operation. Moreover, the behavioral test was performed on rats on the 4th and 28th days after operation. Results Compared with Sham group, the Garcia score of each SAH rat model group decreased significantly on the first day and thereafter increased gradually. However, the recovery rate of each treatment group was higher than the SAH group (no treatment), and the Garcia score of SAH + G1 + HT group was much higher than the SAH group on the seventh day after operation. In addition, each treatment group could obviously reduce the cerebral edema degree of SAH rats, among which rats in SAH + G1 + HT group had lower cerebral edema degree than SAH + G1 group and SAH + HT group. Behavioral test results showed that the combination of GPR30 agonist G1 and HT markedly improved the learning and memory ability of SAH rats, alleviated their anxiety‐ and emotion‐related behavior, and enhanced their social interaction. Conclusion GPR30 agonist G1 combined with HT reduces cognitive impairment and anxiety‐like behavior in rats with SAH.

Abstract Glucocorticoid excess adversely affects male reproduction. This study evaluates effects of pharmacological doses of niacin on testicular structure and function in dexamethasone‐treated rats. Adult rats (48) were randomly assigned to 6 equal groups: (1) Negative control (NC): normal rats; (2) Positive control (PC): dexamethasone at 7 mg/kg/day by intraperitoneal injections for 7 days; groups 3–6 (N50, N100, N200, and N400): dexamethasone and concomitant treatment with niacin at 50, 100, 200, and 400 mg/kg/day by oral gavages. Testicular weight and volume of PC rats were significantly lower than the NC group (p

Abstract The cardiovascular system is primarily controlled by the autonomic nervous system, and any changes in sympathetic or parasympathetic activity also have an impact on myocardial activity. Heart rate variability (HRV) is a readily available metric used to assess heart rate control by the autonomic nervous system. HRV can provide information about neural (parasympathetic, sympathetic, reflex) and humoral (hormones, thermoregulation) control of myocardial activity. Because there are no relevant reference values for HRV parameters in rats in the scientific literature, all experimental results are only interpreted on the basis of changes from currently measured control or baseline HRV values, which are, however, significantly different in individual studies. Considering the significant variability of published HRV data, the present study focused primarily on comparing control or baseline HRV values under different conditions in in vivo experiments involving rats. The aim of the study was therefore to assess whether there are differences in the starting values before the experiment itself.

Abstract The present study assessed a complex of biochemical parameters at the anaerobic threshold (AT) in untrained male Wistar rats with different times to exhaustion (Tex) from swimming. The first group of rats was randomly divided into six subgroups and subjected to a swimming test to exhaustion without a load or with a load of 2%–10% of body weight (BW). In the first group, we established that for untrained rats, the load of 4% BW in the swimming to exhaustion test was optimal for endurance assessment in comparison with other loads. The second group of rats went through a preliminary test with swimming to exhaustion at 4% BW and was then divided into two subgroups: long swimming time (LST, Tex > 240 min) and short swimming time (SST, Tex

Abstract Background Jintiange capsule is composed of bionic tiger bone powder and has similar ingredients to natural tiger bone. Objective To characterize the subacute toxicities of Jintiange capsule in rats and beagle dogs for preclinical safety assessment. Methods Suspensions of Jintiange capsule were given via gastric lavage over a 26‐week period at low (500 mg/kg), mid (1500 mg/kg) and high doses (4000 mg/kg) in SD rats. Beagles were given by gastric lavage of suspensions of Jintiange capsule once daily for 6 days per week for 39 weeks at low (300 mg/kg), mid (900 mg/kg) or high dose (2000 mg/kg). Results Repeated gastric lavages of suspensions of Jintiange capsule at doses from 500 to 4000 mg/kg over 26 weeks caused no significant toxicity (No Observed Adverse Effect Level, NOAEL) in rats. In addition, repeated gastric lavages of suspensions of Jintiange capsule at doses from 300 to 2000 mg/kg over 39 weeks caused NOAEL in beagles. Conclusions Jintiange capsule was safe in rats at a dose 66.7 times the clinically recommended dose and in beagles at 33.3 times the clinically recommended dose. Our subacute toxicity studies in rats and beagles demonstrated no apparent overall toxicities including haematotoxicities, hepatotoxicities and renal toxicities.

Abstract The study examines the influence of three types of hypercaloric diets on metabolic parameters, inflammatory markers, and oxidative stress in experimental model. Male Wistar rats (n = 40) were randomized in control (C), high‐sucrose (HS), high‐fat (HF), and high‐fat with sucrose (HFHS) for 20 weeks. Nutritional, metabolic, hormonal, and biochemical profiles, as well as histological analysis of adipose and hepatic tissues were performed. Inflammation and oxidative stress were determined. HF model caused obesity and comorbidities as glucose intolerance and arterial hypertension. In relation to hormonal and biochemical parameters, there was no significant difference between the groups. All groups showed increased deposition of fat droplets in the hepatic tissue, even though adipocyte areas were similar. Biomarkers of oxidative stress in serum and adipose tissues were similar among the groups. HF model was effective in triggering associated obesity and comorbidities in male rats, but all hypercaloric diets were unable to promote oxidative stress and inflammation.

Ischemia-reperfusion (I/R) injury is a significant clinical problem impacting the heart and other organs, such as the kidneys and liver. This study explores the protective effects of oxycodone on myocardial I/R injury and its underlying mechanisms. Using a myocardial I/R model in Sprague-Dawley (SD) rats and an oxygen-glucose deprivation/reoxygenation (OGD/R) model in H9c2 cells, we administered oxycodone and inhibited AMP-activated protein kinase (AMPK) with Compound C (C.C). Our results showed that oxycodone significantly reduced lactate dehydrogenase (LDH) release and reactive oxygen species (ROS) production while stabilizing mitochondrial membrane potential (MMP). Western blot and RT-qPCR analyzes confirmed that oxycodone enhances AMPK phosphorylation and upregulates the expression of Silent Information Regulator 1 (SIRT1) and Peroxisome Proliferator-Activated Receptor γ Coactivator 1α (PGC-1α), thereby protecting myocardial cells. These findings suggest that oxycodone exerts significant protective effects against I/R injury by activating the AMPK pathway, offering new potential therapeutic targets for myocardial protection., (© 2025 The Author(s). Journal of Biochemical and Molecular Toxicology published by Wiley Periodicals LLC.)

Lycopene (LYC) is an extremely powerful antioxidant with the potential to treat a range of diseases and to inhibit ferroptosis. This research aims to elucidate how LYC impacts polycystic ovarian syndrome (PCOS) and the action mechanisms. A PCOS rat model was constructed by injecting DHEA. Different doses of LYC were injected intraperitoneally in PCOS rats, the estrous cycle was recorded. The histopathological damage of ovary in PCOS rats was observed by HE staining, testosterone (T), estradiol (E2), luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels were examined by ELISA kits. Transmission electron microscopy, prussian blue staining, biochemical kits to determine ferroptosis. Immunohistochemistry and Western blot to assess the levels of ferroptosis-related and AMPK/Nrf2 pathway-related proteins to explore whether LYC affects ferroptosis in PCOS through this pathway. PCOS rats had significantly higher body weights, ovarian weights and ovarian indices, and disorganized estrous cycles, which were dose-dependently ameliorated by LYC. In addition, LYC significantly ameliorated the histopathological damage of ovary in PCOS rats and restored the normal secretion of T, E2, LH, and FSH. LYC attenuates iron deposition in PCOS ovarian tissues, reduces iron and ROS levels, and inhibits ferroptosis. Notably, LYC activated the AMPK/Nrf2 pathway, and AMPK inhibitor intervention attenuated the therapeutic effect of LYC in PCOS rats, suggesting that LYC acts through the AMPK/Nrf2 pathway. LYC attenuates estrous cycle disruption, ameliorates pathological impairments, and inhibits ferroptosis in PCOS rats by modulating the AMPK/Nrf2 pathway., (© 2025 Wiley Periodicals LLC.)

Gualou-Xiebai-Banxia (GXB) decoction shows potential for treating myocardial ischemia (MI), although its underlying mechanism is not fully understood. In this study, a multimodal metabolomics approach, combining gas chromatography-mass spectrometry (GC-MS) and 1 H-NMR, was employed to investigate the cardioprotective effects of GXB in a rat model of myocardial ischemia induced by ligation. ELISA assays and HE staining demonstrated that GXB effectively reduced myocardial injury, oxidative stress markers, and myocardial fibrosis. Orthogonal partial least-squares discriminant analysis identified 62 biomarkers, 20 of which were confirmed using standard compounds. The GC-MS method showed excellent linearity across a wide concentration range (0.004-29.7 μg/mL, R 2  > 0.9995), with intra- and inter-day precision RSD values below 4.72% and 4.96%, respectively. Method recoveries ranged from 95.40% to 104.83%, with RSD values under 4.84%. Pathway enrichment analysis revealed that GXB decoction alleviates myocardial ischemia-induced damage primarily by modulating energy metabolism and branched-chain amino acid metabolism. These findings provide valuable support for the clinical application of GXB decoction in treating myocardial ischemia., (© 2025 John Wiley & Sons Ltd.)

Choerospondias axillaris is a medicinal plant used for treating coronary heart disease (CHD) due to its broad spectrum of anti-inflammatory activities. Cyclooxygenase 2 (COX-2) and lipoxygenase 5 (5-LOX) were immobilized on magnetic nanoparticles for selective ligand-extraction of these two enzymes present in C. axillaris. Sixteen extracted components were identified and analyzed using ultraperformance liquid chromatography plus Q-Exactive Orbitrap tandem mass spectrometry (UHPLC-Q-ExactiveOrbitrap-MS/MS), including flavonoids Curcumin and Epicatechin. The metabolization of the aforementioned 16 components in rats with acute blood stasis indicated a solid pharmacodynamic foundation. This method offers an effective approach for conducting basic research on the enrichment, analysis, and efficacy evaluation of active components in complex natural products., (© 2025 John Wiley & Sons Ltd.)

Colorectal cancer (CRC) represents a significant global health challenge, with approximately 1.8 million new cases diagnosed annually and a mortality toll exceeding 881,000 lives each year. This study aimed to evaluate the chemoprotective efficacy of Cyanidin-3-glucoside (C3G) in a rat model of CRC induced by 1,2-dimethylhydrazine (DMH). Rats were stratified into groups and administered C3G at doses of 10 and 15 mg/kg following DMH exposure to initiate CRC. Key parameters, including organ weights, tumor burdens, and biochemical markers, were meticulously assessed. Administration of C3G significantly restored body weight while reducing the weights of colon and spleen tissues. Moreover, C3G treatment substantially suppressed tumor incidence and weight in DMH-induced CRC rats. Biochemical analysis revealed that C3G markedly reduced levels of CFA, CA19.9, LDH, and nitric oxide (NO). It also modulated lipid profiles, antioxidant activities, and the expression of both Phase I and II enzymes. Inflammatory mediators, including TNF-α, IL-1β, IL-1α, IL-2, IL-4, IL-6, IL-10, IL-12, and IL-17, were significantly downregulated. Notably, C3G inhibited inflammatory markers such as COX-2, PGE2, iNOS, and NF-κB while promoting Caspase-3, -6, and -9 activity. Furthermore, it regulated the Bax/Bcl-2 apoptotic axis, reducing the Bcl-2/Bax ratio. Cyanidin-3-glucoside demonstrated potent chemopreventive effects against colorectal cancer in this experimental model. Its mechanism of action is likely mediated through modulation of NF-κB and the Bcl-2/Bax/Caspase pathway, suggesting its potential as a therapeutic agent in CRC management., (© 2025 Wiley Periodicals LLC.)

Panax notoginseng (P. notoginseng) is one of the most famous natural medicines and widely used to promote blood circulation in health care. However, the active component group of P. notoginseng for activating blood is not clear. We aim to screen and validate the pharmacodynamic component group (PCG), which could exert the same blood-activating effect as P. notoginseng. To clarify the active components, the chemical components were determined by liquid chromatography-tandem mass spectrometry, and the fingerprint of P. notoginseng was established. Twenty candidate active monomers were selected through the spectrum-effect relationship analysis. Eleven active monomers, including Ginsenoside Rg1, Rb1, Rd, F1, Rh1, Rg2, Rb2, Rg3, and Rk1 and Notoginsenoside R1 and R2, were screened out as the PCG through validation by platelet aggregation test. Among them, the antiplatelet aggregation activity of Ginsenoside Rh1 was directly confirmed for the first time. The active component group could exert similar efficacy to the P. notoginseng extract in vitro and in vivo through the validation of in vitro platelet aggregation test and the rats with cerebral ischemia. This study laid the foundation for the quality evaluation of P. notoginseng and provided a reference for the research on the material basis of the pharmacodynamics of other Chinese herbs., (© 2025 John Wiley & Sons Ltd.)

Vodobatinib is a Bcr-Abl 1 inhibitor, currently entering into Phase 2 clinical trials as a potential drug to treat glioblastoma patients. In the present work, a validated high-performance liquid chromatography (HPLC) detection method for the quantification of vodobatinib in rat plasma was established. Sample preparation involved liquid-liquid extraction method. The chromatographic separation of vodobatinib and the IS was accomplished on an XBridge C 18 column using 10 mM ammonium acetate and acetonitrile in gradient condition. The flow-rate and injection volume were 0.8 mL/min and 20 μL, respectively. The eluent was detected at 310 nm. Vodobatinib and the IS eluted at 7.5 and 5.9 min, respectively, with a total run time of 10 min. Validation tests were performed according to FDA requirements. The established method showed good linearity in the range of 50.5-6534 ng/mL. Interday and intraday coefficients of variations were < 5.11%. Stability studies, dilution integrity, and incurred sample reanalysis met the acceptance criteria. Subsequently, the validated HPLC method was used to study the disposition kinetics of vodobatinib in rats., (© 2025 John Wiley & Sons Ltd.)

Background and Purpose: The dorsal hippocampus (dHIP) is pivotal for learning, memory, and defensive responses. Transient receptor potential vanilloid type 1 (TRPV1) receptors in the dHIP modulate contextual fear conditioning by triggering a cascade involving glutamate release, nitric oxide (NO) formation and cyclic guanosine monophosphate (cGMP) production. The present study investigated the involvement of dHIP TRPV1 receptors and their interaction with the glutamate/NO/cGMP signalling pathway in modulating the expression of contextual fear conditioning (CFC)., Experimental Approach: Male Wistar rats were submitted to an aversive contextual conditioning session and, 48 h later, were re-introduced to the same aversive environment where the freezing response and autonomic activity (evidenced by increased arterial pressure and heart rate and a decrease in tail temperature) were measured., Key Results: The results demonstrated that the TRPV1 antagonist 6-I-CPS in dHIP reduced the expression of CFC, whereas the agonist capsaicin had the opposite effect. Furthermore, dHIP pre-treatment with an NMDA receptor antagonist (AP7), neuronal NO synthase inhibitor (N-propyl-L-arginine), NO scavenger (c-PTIO) or guanylate cyclase inhibitor (ODQ) attenuated capsaicin-induced increases in CFC. Finally, we observed that re-exposure to the aversive chamber increased dHIP NO levels in conditioned animals compared with a non-conditioned group, which was prevented by the administration of the TRPV1 antagonist, 6-I-CPS., Conclusion and Implications: Our study revealed that TRPV1 receptors in the dHIP play a crucial role in modulating contextual fear expression by acting through the NMDA receptor/NO/cGMP signalling pathway, providing important insights into the underlying mechanisms and potential therapeutic avenues associated with these pathways., (© 2024 British Pharmacological Society.)

Magnetic resonance spectroscopic imaging (MRSI) enables the simultaneous noninvasive acquisition of MR spectra from multiple spatial locations inside the brain. Although 1 H-MRSI is increasingly used in the human brain, it is not yet widely applied in the preclinical setting, mostly because of difficulties specifically related to very small nominal voxel size in the rat brain and low concentration of brain metabolites, resulting in low signal-to-noise ratio (SNR). In this context, we implemented a free induction decay 1 H-MRSI sequence ( 1 H-FID-MRSI) in the rat brain at 14.1 T. We combined the advantages of 1 H-FID-MRSI with the ultra-high magnetic field to achieve higher SNR, coverage, and spatial resolution in the rat brain and developed a custom dedicated processing pipeline with a graphical user interface for Bruker 1 H-FID-MRSI: MRS4Brain toolbox. LCModel fit, using the simulated metabolite basis set and in vivo measured MM, provided reliable fits for the data at acquisition delays of 1.30 ms. The resulting Cramér-Rao lower bounds were sufficiently low (< 30%) for eight metabolites of interest (total creatine, N-acetylaspartate, N-acetylaspartate + N-acetylaspartylglutamate, total choline, glutamine, glutamate, myo-inositol, and taurine), leading to highly reproducible metabolic maps. Similar spectral quality and metabolic maps were obtained with one and two averages, with slightly better contrast and brain coverage due to increased SNR in the latter case. Furthermore, the obtained metabolic maps were accurate enough to confirm the previously known brain regional distribution of some metabolites. The acquisitions proved high reproducibility over time. We demonstrated that the increased SNR and spectral resolution at 14.1 T can be translated into high spatial resolution in 1 H-FID-MRSI of the rat brain in 13 min using the sequence and processing pipeline described herein. High-resolution 1 H-FID-MRSI at 14.1 T provided robust, reproducible, and high-quality metabolic mapping of brain metabolites with minimal technical limitations., (© 2024 John Wiley & Sons Ltd.)

Background and Purpose: Therapeutic hypothermia (TH) has become the standard care to reduce morbidity and mortality in neonates affected by moderate-to-severe hypoxic-ischaemic encephalopathy (HIE). Despite the use of TH for HIE, the incidence of mortality and disabilities remains high., Experimental Approach: Nerve growth factor (NGF) is a potent neurotrophin, but clinical use is limited by its pain eliciting effects. CHF6467 is a recombinant modified form of human NGF devoid of algogenic activity (painless NGF)., Key Results: In rodent hippocampal slices exposed to oxygen and glucose deprivation, CHF6467 protected neurons from death and reverted neurotransmission impairment when combined with hypothermia. In a model of rat neonatal HIE, intranasal CHF6467 (20 μg kg -1 ) significantly reduced brain infarct volume versus vehicle when delivered 10 min or 3 h after the insult. CHF6467 (20 and 40 μg kg -1 , i.n.), significantly decreased brain infarct volume to a similar extent to TH and when combined, showed a synergistic neuroprotective effect. CHF6467 (20 μg kg -1 , i.n.) per se and in combination with hypothermia reversed locomotor coordination impairment (Rotarod test) and memory deficits (Y-maze and novel object recognition test) in the neonatal HIE rat model. Intranasal administration of CHF6467 resulted in meaningful concentrations in the brain, blunted HIE-induced mRNA elevation of brain neuroinflammatory markers and, when combined to TH, significantly counteracted the increase in plasma levels of neurofilament light chain, a peripheral marker of neuroaxonal damage., Conclusion and Implications: CHF6467 administered intranasally is a promising therapy, in combination with TH, for the treatment of HIE., (© 2024 British Pharmacological Society.)

Background and Purpose: In male rats, the serotonergic system modulates sympathetic outflow at vascular levels, causing sympatho-inhibition and sympatho-excitation, mainly via 5-HT 1D/1A and 5-HT 3 receptors, respectively. However, sex influence on vascular serotonergic regulation has not yet been elucidated. This study aimed to analyse the 5-HT sympatho-modulatory role in female rats, characterising the 5-HT receptors involved., Experimental Approach: Female Wistar (14- to 16-week-old) rats were prepared for sympathetic stimulation. Mean blood pressure (MBP) and heart rate (HR) were continuously measured. Vasopressor responses were obtained by electrical stimulation of the sympathetic outflow (0.1-5 Hz) or i.v. noradrenaline (0.01-0.5 μg·kg -1 ). 5-HT-related drug effects on adrenergic system were determined. Age-matched male rats were used as control., Key Results: Basal MBP in females was lower than in male rats, whereas electrical-induced increases in MBP were similar. In females, 5-HT exerted a dose-dependent inhibition on the sympathetic-evoked vasoconstrictions, that was reproduced by some agonists; 5-CT (5-HT 1/5/7 ) and L-694,247 (5-HT 1D ), whereas the selective 5-HT 2A/2B/2C (α-methyl-5-HT) and 5-HT 3 agonist (1-PBG) increased the electrically-produced vasopressor responses. None of the other drugs tested (targeting 5-HT 1A/1B/1F , 5-HT 2B/2C , 5-HT 4 , 5-HT 5A or 5-HT 7 ) modified these vasoconstrictions. Only 1-PBG (5-HT 3 ) modified the vasoconstrictions induced by exogenous noradrenaline., Conclusions and Implications: In female rats, vascular serotonergic sympatholytic effects are due to prejunctional 5-HT 1D receptor activation, whereas pre and/or postjunctional 5-HT 3 and prejunctional 5-HT 2A receptor activation is involved in the potentiating effect of vascular sympathetic neurotransmission. These findings may open novel sex-differential therapeutic strategies for treating cardiovascular conditions., (© 2024 The Author(s). British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of British Pharmacological Society.)

Background: Painful neuropathy is a pathological condition caused by numerous factors including diabetes, chemotherapy or cancer. ART26.12 is a novel fatty acid-binding protein 5 inhibitor, which our group showed could prevent and treat persistent pain in a preclinical model of oxaliplatin-induced peripheral neuropathy., Methods: In the current study, the efficacy of orally dosed ART26.12 was tested in multiple neuropathy models of different aetiology. Paw withdrawal threshold to von Frey monofilaments and latency to escape a cold plate were used as measurements of mechanical and cold sensitivity., Results: ART26.12 (25 and 50 mg/kg BID), dosed prior to the induction of paclitaxel-induced peripheral neuropathy (PIPN), reversed mechanical allodynia induced by paclitaxel in both male and female rats, and ART26.12 (50 mg/kg BID) prevented the induction of PIPN in female rats. ART26.12 (50 mg/kg BID) also had a protective effect on body weight in the PIPN model. ART26.12 (25 and 100 mg/kg BID) reversed mechanical allodynia when treating established streptozotocin-induced diabetic neuropathy in male rats. In a model of breast cancer-induced bone pain in female rats, ART26.12 (100 mg/kg BID) reversed mechanical allodynia within 1 h of dosing. In the same model, ART26.12 (25 mg/kg BID) reversed mechanical allodynia from day 4 of treatment., Conclusion: Overall, these preclinical data suggest that ART26.12 is a safe and efficacious therapeutic drug for continued development towards the prevention and treatment of peripheral neuropathy., Significance Statement: This work now shows that ART26.12, a novel and selective inhibitor of FABP5, can prevent and treat multiple preclinical models of peripheral neuropathy. Given its excellent safety profile, further work is warranted to develop ART26.12 as a potential therapeutic tool for pain management., (© 2024 The Author(s). European Journal of Pain published by John Wiley & Sons Ltd on behalf of European Pain Federation ‐ EFIC ®.)

Background and Purpose: Botulinum neurotoxin type A1 (BoNT/A) is one of the most potent neurotoxins known. At the same time, it is also one of the safest therapeutic agents used for the treatment of several human disorders and in aesthetic medicine. Notwithstanding great effectiveness, strategies to accelerate the onset and prolong BoNT/A action would significantly ameliorate its pharmacological effects with beneficial outcomes for clinical use., Experimental Approach: Here, we combined BoNT/A with two fast-acting inhibitors of excitation-contraction coupling inhibitors (ECCI), either the μ-conotoxin CnIIIC or dantrolene, and tested the effect of their co-injection on a model of hind-limb paralysis in rodents using behavioural, biochemical, imaging and electrophysiological assays., Key Results: The BoNT/A-ECCI combinations accelerated the onset of muscle relaxation. Surprisingly, they also potentiated the peak effect and extended the duration of the three BoNT/A commercial preparations OnabotulinumtoxinA, AbobotulinumtoxinA and IncobotulinumtoxinA. ECCI co-injection increased the number of BoNT/A molecules entering motoneuron terminals, which induced a faster and greater cleavage of SNAP-25 during the onset and peak phases, and prolonged the attenuation of nerve-muscle neurotransmission during the recovery phase. We estimate that ECCI co-injection yields a threefold potentiation in BoNT/A pharmacological activity., Conclusions and Implications: Overall, our results show that the pharmacological activity of BoNT/A can be combined and synergized with other bioactive molecules and uncover a novel strategy to enhance the neuromuscular effects of BoNT/A without altering the neurotoxin moiety or intrinsic activity, thus maintaining its exceptional safety profile., (© 2024 The Author(s). British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of British Pharmacological Society.)

Lead (Pb) exposure during perinatal development alters testosterone (T) concentrations and delays puberty in children and laboratory rodents. In addition, exposure to the metal during adult life decreases T and libido in men and affects male reproductive behaviour (MRB) in rats. MRB is regulated by various brain nuclei including the medial preoptic area (MPOa) and the medial amygdala (MeA), in which T and oestradiol (E 2 ) act through their respective androgen (AR) and oestrogen (ER) receptors. However, the mechanism by which MRB is affected by Pb exposure is not known. The objectives of the present study were to evaluate whether perinatal Pb exposure affects MRB and the number of cells immunoreactive to AR and ERα in the MPOa and the MeA. Male Wistar rats exposed to Pb (320 ppm) in drinking water from the beginning of pregnancy until weaning were used. The experimental group experienced significant alterations in MRB, an important decrease in T and E 2 concentrations, and a significant increase in Pb concentrations in the blood, MPOa (hypothalamus) and MeA. In addition, in the studied areas the number of cells immunoreactive to AR and ERα, or detected using the Nissl technique, decreased significantly. These results show that perinatal exposure to Pb alters MRB. This event may be related to a decrease in both the concentrations of sex hormones and the number of cells that express their receptors as well as in the neuronal Nissl staining population. This ultimately affects the quality of life of the individual., (© 2024 Company of the International Journal of Experimental Pathology (CIJEP).)

The high mortality rate of chronic heart failure (CHF) makes it a primary battlefield in the field of cardiovascular diseases. Qiangxin Lishui Prescription (QLP) is a traditional Chinese medicine (TCM) prescription used clinically for treating CHF, but its underlying mechanism remains unclear. This study integrated plasma metabolomics, network pharmacology, and experimental validation to reveal the pharmacological effects of QLP and its potential mechanism of anti-CHF. Using ultra-high-performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS/MS), 119 absorbed prototype compounds of QLP were identified from rat plasma. By applying network pharmacology and molecular docking techniques, a QLP absorption components-target-CHF network was constructed. The IL6/JAK/STAT3 signaling pathway is likely critical to QLP's therapeutic effect on CHF. A CHF mouse model was established using aortic ligation surgery to investigate the regulation of the IL6/JAK/STAT3 pathway by QLP in CHF mice. Network pharmacology analysis and in vivo experimental data indicate that QLP alleviates myocardial injury and inflammatory response in CHF mice by modulating the IL6/JAK/STAT3 pathway, significantly improving cardiac function. This presents a promising therapeutic strategy for CHF treatment., (© 2024 John Wiley & Sons Ltd.)

Background and Purpose: Recent research linking choline-containing lipids to degeneration of basal forebrain cholinergic neurons in neuropathological states illustrates the challenge of balancing lipid integrity with optimal acetylcholine levels, essential for memory preservation. The endocannabinoid system influences learning and memory processes regulated by cholinergic neurotransmission. Therefore, we hypothesised that activation of the endocannabinoid system may confer neuroprotection against cholinergic degeneration., Experimental Approach: We examined the neuroprotective potential of sub-chronic treatments with the cannabinoid agonist WIN55,212-2, using ex vivo organotypic tissue cultures including nucleus basalis magnocellularis and cortex and in vivo rat models of specific cholinergic damage induced by 192IgG-saporin. Levels of lipids, choline and acetylcholine were measured with histochemical and immunofluorescence assays, along with [ 35 S]GTPγS autoradiography of cannabinoid and muscarinic GPCRs and MALDI-mass spectrometry imaging analysis. Learning and memory were assessed by the Barnes maze and the novel object recognition test in rats and in the 3xTg-AD mouse model., Key Results: Degeneration, induced by 192IgG-saporin, of baso-cortical cholinergic pathways resulted in memory deficits and decreased cortical levels of lysophosphatidylcholines (LPC). WIN55,212-2 restored cortical cholinergic transmission and LPC levels via activation of cannabinoid receptors. This activation altered cortical lipid homeostasis mainly by reducing sphingomyelins in lesioned animals. These modifications were crucial for memory recovery., Conclusion and Implications: We hypothesise that WIN55,212-2 facilitates an alternative choline source by breaking down sphingomyelins, leading to elevated cortical acetylcholine levels and LPCs. These results imply that altering choline-containing lipids via activation of cannabinoid receptors presents a promising therapeutic approach for dementia linked to cholinergic dysfunction., (© 2024 The Author(s). British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of British Pharmacological Society.)

Background and Purpose: Cisplatin-induced sensorineural hearing loss is a significant clinical challenge. Although the potential effects of brain-derived neurotrophic factor (BDNF) have previously been investigated in some ototoxicity models, its efficacy in cisplatin-induced hearing loss remains uncertain. This study aimed to investigate the therapeutic potential of recombinant human BDNF (rhBDNF) in protecting cells against cisplatin-induced ototoxicity., Experimental Approach: Using an in vivo model of cisplatin-induced hearing loss, we investigated the beneficial effects of transtympanic administration of rhBDNF in a thermogel solution on hearing function and cochlear injury, using electrophysiological, morphological, immunofluorescence and molecular analyses., Key Results: Our data showed that local rhBDNF treatment counteracted hearing loss in rats receiving cisplatin by preserving synaptic connections in the cochlear epithelium and protecting hair cells (HCs) and spiral ganglion neurons (SGNs) against cisplatin-induced cell death. Specifically, rhBDNF maintains the balance of its receptor levels (pTrkB and p75), boosting TrkB-CREB pro-survival signalling and reducing caspase 3-dependent apoptosis in the cochlea. Additionally, it activates antioxidant mechanisms while inhibiting inflammation and promoting vascular repair., Conclusion and Implications: Collectively, we demonstrated that early transtympanic treatment with rhBDNF plays a multifaceted protective role against cisplatin-induced ototoxicity, thus holding promise as a novel potential approach to preserve hearing in adult and paediatric patients undergoing cisplatin-based chemotherapy., (© 2024 The Author(s). British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of British Pharmacological Society.)

Proteolysis targeting chimera (PROTAC) is emerging as a promising medicinal modality, which has aroused widespread interest among the field of pharmaceutical manufacturing in the recent years. ARV-471 is an orally active PROTAC estrogen receptor degrader against breast cancer, which leads to the ubiquitylation and subsequent degradation of estrogen receptors via the proteasome. In this study, we developed a highly sensitive liquid chromatography tandem mass spectrometry method (LLOQ = 0.5 ng/mL) for the measurement of ARV-471 in rat plasma. The acetonitrile precipitated sample was separated on ACQUITY BEH C 18 column using acetonitrile-0.1% formic acid as mobile phased with gradient elution. Multiple reactions monitoring in positive ESI mode was employed for the quantification of ARV-471 (m/z 724.4 → 396.2). The assay showed good linearity over the concentration range of 0.5-1000 ng/mL with correlation coefficient > 0.996. The assay was validated according to FDA guidance, and all the validation parameters were within the predefined acceptance criteria. After validation, the assay was applied to the pharmacokinetic study of ARV-471 in rats. Additionally, the metabolites in rat plasma were identified using liquid chromatography-high resolution mass spectrometry. Four metabolites were identified and characterized. Hydrolysis, glucuronidation and deamination were the main metabolic pathways of ARV-471 in rats., (© 2024 John Wiley & Sons Ltd.)

The aim of this study was to investigate the potential mechanism of Lu-Jiao Fang (LJF) inhibiting endothelial-to-mesenchymal transition (EndMT) in pressure overload-induced cardiac fibrosis. Pharmacokinetic behaviors of the ingredients of LJF were evaluated by LC-MS/MS analysis. Then putative pathways by which LJF regulates EndMT were analyzed by network pharmacology and verified in transverse aortic constriction-induced cardiac fibrosis rats. Loganin, morroniside, salidroside, isopsoralen, and psoralen showed higher plasma, left and right ventricular C max and AUC 0-t values than hesperidin, specnuezhenide, and icariside II. Twenty-four potential targets related to EndMT were identified, which were mainly involved in relaxin signaling pathway. AKT1, TP53, MMP9, HIF1A, Snail1, and MMP2 were key therapeutic targets in protein-protein interaction network. LJF reversed cardiac dysfunction, left ventricular dilation, and fibrosis and significantly downregulated collagen type I and III and EndMT regulators (Snail1 and Twist1) mRNA expression. In relaxin signaling pathway, the RXFP1 protein expression increased by 22.52%, and the protein phosphorylation of Smad2 and Smad3 decreased by 33.52% and 12.79%, in response to the treatment with LJF. This study initially revealed the EndMT inhibition effects and molecular mechanisms of LJF in cardiac fibrosis, providing a reference basis for the promotion of LJF in the clinic., (© 2025 John Wiley & Sons Ltd.)

Background and Purpose: Opioid analgesics can alleviate ischaemia/reperfusion (I/R) injury in chronic heart failure. However, the underlying mechanisms and targets remain unknown. Here, we investigate if caveolin-3 (Cav3) interacts with μ opioid receptors and if Cav3-μ receptor interactions play a role in morphine-induced cardioprotection in failing hearts., Experimental Approach: Cav3 and μ receptor proteins in human and rat heart tissue were determined by western blot, immunofluorescence and co-immunoprecipitation. Methyl-β-cyclodextrin (MβCD), a destroyer of caveolae, and AAV-Cav3 shRNA were used to reduce Cav3 expression in failing rat hearts. CTOP, a specific μ antagonist, was administrated before morphine preconditioning in perfused failing heart models of myocardial I/R injury., Key Results: Levels of Cav3 and μ receptor proteins were significantly higher in human and rat myocardial tissues with heart failure than in control tissues. Cav3 and μ receptor expression levels were positively correlated with disease severity. The signal of the cardiac Cav3 protein was colocalized with μ receptor in both the human and rat heart sections. Disruption of caveolae in the failing heart by either MβCD or AAV-Cav3 shRNA significantly inhibits morphine-induced phosphorylation of ERK1/2 and cardioprotection. Administration of CTOP substantially reduced Cav3 expression and morphine-induced cardioprotective effect in heart failure., Conclusion and Implications: Our data suggest that up-regulation of the Cav3/μ receptor complex is critical for morphine protection of the failing heart against I/R injury by regulating the ERK1/2 pathway. The activated Cav3/μ receptor complex is an understudied therapeutic target for opioid treatment of heart failure and ischaemic insult., (© 2024 British Pharmacological Society.)

Background: Ethnomedicine exhibits potential in developing affordable effective antidiabetic agents. Aim: This work aimed to explore the antidiabetic properties of Aloe niebuhriana latex extract both in vivo, utilizing alloxan-induced diabetic rats, and in vitro, through α -amylase enzyme testing. Additionally, it sought to formulate optimal effervescent granules derived from the extract. Methods: The α -amylase inhibition assay was performed using the α -amylase kit using biochemical analyzers. Experimental diabetes was induced in animals with alloxan. On Day 14 postdiabetes induction, body weight, fasting blood glucose, and lipid profile parameters were determined. Also, six effervescent granule preparations of the extract were formulated using wet granulation. Based on its physical and organoleptic properties, a formulation was selected and optimized. Results: The extract displayed modest α -amylase inhibition, with an IC 50 value of 439.2  μ g/mL. Both doses of A. niebuhriana extract (200 and 400 mg/kg) significantly reduced blood glucose level compared to their respective Day 1 levels ( p < 0.001). Moreover, the extract at a dose of 400 mg/kg significantly normalized lipid profile compared to the diabetic control groups ( p < 0.05 - 0.001). Six formulations containing the extract were prepared (F1-F6), and F6 containing 200 mg of the extract was selected for optimization due to its favorable odor, taste, foaming, and effervescent properties, high solubility, and absence of turbidity and adhesion. The formulated F6 granules successfully met the quality parameters assessed including flow time, pH effervescent time, angle of repose, bulk density, tapped density, Carr's index, and Hausner's ratio. Conclusion: This study highlights the antidiabetic potential of A. niebuhriana latex extract, potentially attributed to its hypolipidemic, hypoglycemic, and α -amylase inhibitory effects. The successful formulation and evaluation of the extract as effervescent granules suggest its potential as an antidiabetic drug., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2025 Bushra Abdulkarim Moharram et al. The Scientific World Journal published by John Wiley & Sons Ltd.)

Amnesia is a memory disorder marked by the inability to recall or acquire information. Hence, drugs that also target the neurogenesis process constitute a hope to discover a cure against memory disorders. This study is aimed at evaluating the antiamnesic and neurotrophic effects of the aqueous extract of Parkia biglobosa ( P. biglobosa ) on in vivo and in vitro models of excitotoxicity. For the in vivo study, 42 adult male rats were divided into six groups of seven rats each and treated daily for 30 days as follows: normal control group (distilled water, 10 mL/kg, po), negative control group (distilled water, 10 mL/kg, po), positive control group (piracetam, 200 mg/kg, po), and 03 test groups (extract, 44, 88, and 176 mg/kg, po). Scopolamine (0.5 mg/kg, ip) was administered once daily, 45 min after these treatments, for 14 days, except in the normal control group. The animals were then subjected to short-term memory (new object recognition and T-maze) and long-term memory (radial arm maze) tests for 15 following days. Animals were then euthanized, and biochemical analyses (neurotransmitters, oxidative status, and neuroinflammation) were performed in the prefrontal cortex, hippocampus, and serum. Histological analysis of these organs was also carried out. In the in vitro study, the effect of the extract (5, 10, 19, 40, 77, 153, 306, 615, 1225, and 2450  μ g/mL) was assessed on the viability of primary cortical neurons exposed to L-glutamate (0.1 mg/mL). Scopolamine induced memory impairment and increased oxidative stress, neuroinflammation, and neuronal loss. P. biglobosa extract (44 mg/kg) reduced ( p < 0.001) short- and long-term memory deficit. It also increased ( p < 0.01) the concentration of acetylcholine, reduced ( p < 0.001) that of malondialdehyde, and limited ( p < 0.001) neuroinflammation and neuronal loss ( p < 0.001). In addition, the extract (2450  μ g/mL) increased ( p < 0.001) the percentage of viable cells. These results suggest that the extract has effects on amnesia and neurogenesis. These effects seem to be mediated by antioxidant and anti-inflammatory modulations., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2025 Antoine Kavaye Kandeda et al. Behavioural Neurology published by John Wiley & Sons Ltd.)

Bisphenol A (BPA), an environmental endocrine disrupting chemical, is one of the most widely used chemicals in the world and is widely distributed in the external environment, specifically in food, water, dust, and soil. BPA exposure is associated with abnormal cognitive behaviors. However, the underlying mechanism remains unclear. In this study, pregnant female Sprague Dawley rats were orally exposed to BPA at a low dose of 0, 0.04, 0.4, or 4 mg per kg·of body weight per day from embryonic Day 0 (ED 0) to postnatal Day 21 (PND 21). Spatial learning and memory were measured via a Morris water maze test on PND 22. PI3K/Akt/mTOR signaling pathway protein expression was detected in the hippocampi of male offspring using a western blot. The water maze test demonstrated that BPA exposure considerably reduced the learning and memory capacities of the male offspring exposure groups when compared to the control group. The male offspring rats' latency to escape increased significantly, the time taken to traverse a platform reduced, and latency to find a hidden platform showed an increasing trend. Meanwhile, maternal exposure to BPA downregulated the expression of PI3K/Akt/mTOR/p70S6K pathway in the hippocampi of the offspring. Moreover, BPA exposure improved the GSK3β and phosphorylated tau protein (T231) levels, increased malondialdehyde levels, and activated caspase-3 expression in the hippocampi of the male offspring rats. Taken together, these findings indicate that maternal exposure to BPA causes learning and memory impairment and that the PI3K/Akt/mTOR pathway participates in the mechanism of BPA-induced neurocognitive decline., (© 2024 Wiley Periodicals LLC.)

Phospholipase A2 receptor 1 (PLA2R1) exists in many animals and plays an important role in membranous nephropathy. In this study, we aimed to evaluate a PLA2R1 knock-in rat model with repaired kidney function to study the molecular mechanisms of membranous nephropathy. We constructed the PLA2R1 knockout [PLA2R1(-)] model and PLA2R1 knock in [PLA2R1(+)] model in rats. Consistent complement C3 and IgA expression was confirmed through colocalization studies. Urinary biochemical indicators were performed using Automatic Biochemistry Analyzer. The complement C3, IgG, and Nephrin were detected by immunofluorescence assay. The expression levels of complement C3, IgA, and PLA2R1 were detected by western blot. The differential expression proteins (DEPs) between control and PLA2R1(+) models were detected by liquid chromatography with tandem mass spectrometry. The PLA2R1(-) model showed proteinuria, complement C3 aggregation, and IgA and IgG deposition in the glomerulus. Comparing with the PLA2R1(-) model, the PLA2R1(+) model, the deposition of complement C3 and IgA in the glomerulus did not completely disappear, and IgG expression weakened. Moreover, the absolute value of urinary protein was much lower in the PLA2R1(+) model than in the PLA2R1(-) model, and some of the humanized PLA2R1 gene fragments repaired some of the kidney functions. Humanized PLA2R1-insertion in rats can repair part of the renal function and reduce proteinuria, which will help in studying the molecular mechanisms of membranous nephropathy, as well as the entire membranous nephropathy-related system and complement activation signaling pathway., (© 2024 Wiley Periodicals LLC.)

Ovarian ischemia is a pathological condition that usually occurs due to ovarian torsion, resulting in the interruption of blood supply to the ovaries and oxygen deficiency. Silymarin (SLM) is a flavonoid complex of plant origin with pharmacological properties such as antioxidant, anti-inflammatory, and antiapoptotic effects. In this study, we investigated the effects of SLM through different pathways in rats subjected to experimental ovarian ischemia/reperfusion (I/R). Female Wistar rats were divided into five groups: Control, SLM (50 mg/kg), I/R, I/R + SLM25 (25 mg/kg), and I/R + SLM50 (50 mg/kg). SLM was given orally for 7 days, followed by ischemia (2 h) and reperfusion (2 h) on day 8. Biochemical (MDA, GSH, SOD, CAT, GPx) and histological (H&E, Ki-67 IHC) analyses were performed. Also, molecular (qRT-PCR) analyses were performed to evaluate oxidative stress, inflammation, apoptosis, and Wnt signaling. I/R increased MDA and NO levels in ovarian tissue while decreasing SOD, CAT, GPx, and GSH. Antioxidant defense genes (Nrf-2, HO-1, NQO1) were suppressed, and inflammation markers (NF-ĸB, IL-1β, TNF-α) along with apoptotic markers (Bax, Caspase-3) were elevated, while Bcl-2 decreased. The Wnt signaling pathway was inhibited, particularly at Wnt-3A, LRP5, Dvl-2, and Cyclin-1, reducing Ki-67 protein levels and IHC positivity. Silymarin has shown a therapeutic effect on ovarian ischemia reperfusion injury with its antioxidant, antiapoptotic and anti-inflammatory effects and cell cycle regulatory activity., (© 2025 The Author(s). Journal of Biochemical and Molecular Toxicology published by Wiley Periodicals LLC.)

Acute myocardial infarction (AMI) is a leading cause of mortality among cardiovascular diseases, yet effective therapies for AMI are limited. Previous studies have suggested cardioprotective effects of columbianadin (CBN), but its specific role in AMI and the underlying mechanisms remain unclear. This study aims to investigate whether CBN influences AMI and to elucidate the underlying mechanisms. We conducted a network pharmacology analysis to investigate the relationship between CBN and AMI. The AMI model was established by ligating the left anterior descending (LAD) artery in C57BL/6J mice, which were subsequently administered CBN. Hypoxic H9c2 cells were utilized to evaluate the effects of CBN in vitro. Our study revealed that CBN treatment significantly reduced myocardial infarction in AMI mice. It enhanced mitochondrial function and suppressed autophagy flux in hypoxic H9c2 cells. Furthermore, CBN downregulated the expression of LC3, Beclin1, and Atg 5 genes and proteins. In response to CBN treatment, the phosphorylation levels of PI3K, Akt, and mTOR increased. Notably, RAPA attenuated the protective effect of CBN in enhancing the survival of hypoxic H9c2 cells and abolished its regulation of autophagy-related proteins via the PI3K/Akt/mTOR signaling pathway. In conclusion, CBN reduces myocardial damage by suppressing autophagy via the PI3K/Akt/mTOR signaling pathway in AMI mice and hypoxic H9c2 cells., (© 2024 John Wiley & Sons Ltd.)

Over 90% of protein degradation in eukaryotic cells occurs through the ubiquitin-proteasome system (UPS). In this system, the ubiquitin protein can bind to a substrate on its own or it can form a chain with multiple ubiquitin molecules in a process called polyubiquitination. There are 8 different sites on ubiquitin at which polyubiquitin chains can be formed, the second most abundant of which, lysine-63 (K63), is independent of the degradation process, though this mark has rarely been studied in the brain or during learning-dependent synaptic plasticity. Recently, we found that knockdown of K63 polyubiquitination in the amygdala selectively impaired contextual fear memory formation in female, but not male, rats. It is unknown, however, whether the sex-specific requirement of K63 polyubiquitination occurs in other brain regions that are required for contextual fear memory formation, including the hippocampus. Here, we found that CRISPR-dCas13-mediated knockdown of K63 polyubiquitination in the hippocampus significantly enhanced contextual fear memory in both male and female rats, a result that is in striking contrast to what we observed in the amygdala for both sex-specificity and directionality. Using unbiased proteomics, we found that following fear conditioning K63 polyubiquitination was primarily decreased at target proteins in the hippocampus of both males and females. Importantly, the target proteins and downstream functional pathways influenced by K63 polyubiquitination changes diverged significantly by sex. Together, these data suggest that unlike what we previously reported in the amygdala, decreases in K63 polyubiquitination in the hippocampus are a critical regulator of memory formation in the hippocampus of both males and females., (© 2024 Wiley Periodicals LLC.)

Background and Purpose: Cardioprotective miRNAs (protectomiRs) are promising therapeutic tools. Here, we aimed to identify protectomiRs in a translational porcine model of acute myocardial infarction (AMI) and to validate their cardiocytoprotective effect., Experimental Approach: ProtectomiR candidates were selected after systematic analysis of miRNA expression changes in cardiac tissue samples from a closed-chest AMI model in pigs subjected to sham operation, AMI and ischaemic preconditioning, postconditioning or remote preconditioning, respectively. Cross-species orthologue protectomiR candidates were validated in simulated ischaemia-reperfusion injury (sI/R) model of isolated rat ocardiomyocytes and in human AC16 cells as well. For miR-450a, we performed target prediction and analysed the potential mechanisms of action by GO enrichment and KEGG pathway analysis., Key Results: Out of the 220 detected miRNAs, four were up-regulated and 10 were down-regulated due to all three conditionings versus AMI. MiR-450a and miR-451 mimics at 25 nM were protective in rat cardiomyocytes, and miR-450a showed protection in human cardiomyocytes as well. MiR-450a has 3987 predicted mRNA targets in pigs, 4279 in rats and 8328 in humans. Of these, 607 genes are expressed in all three species. A total of 421 common enriched GO terms were identified in all three species, whereas KEGG pathway analysis revealed 13 common pathways., Conclusion and Implications: This is the first demonstration that miR-450a is associated with cardioprotection by ischaemic conditioning in a clinically relevant porcine model and shows cardiocytoprotective effect in human cardiomyocytes, making it a promising drug candidate. The mechanism of action of miR-450a involves multiple cardioprotective pathways., Linked Articles: This article is part of a themed issue Non-coding RNA Therapeutics. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v182.2/issuetoc., (© 2024 The Author(s). British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of British Pharmacological Society.)

For most of my career, I focused on understanding how and where spatial context, the place where things happen, is represented in the brain. My interest in this began in the early 1990's, during my postdoctoral training with David Amaral, when we defined the rodent homolog of the primate parahippocampal cortex, a region implicated in processing spatial and contextual information. We parceled out the caudal portion of the rat perirhinal cortex (PER) and called it the postrhinal cortex (POR). In my own lab at Brown University, I continued to study the anatomy of the PER, POR, and entorhinal cortices. I also began to characterize and differentiate the functions of these regions, particularly the newly defined POR and the redefined PER. Our electrophysiological and behavioral evidence supports a view of POR function that aligns with our anatomical evidence. Briefly, the POR integrates object and feature information from the PER with spatial information from the retrosplenial, posterior parietal, and secondary visual cortices and the pulvinar and uses this information to represent specific environmental contexts, including the spatial arrangement of objects and features within each context. In addition to maintaining a representation of the current context, the POR plays an attentional role by continually monitoring the context for changes and updating the context representation when changes occur. This context representation is accessible to other regions for cognitive processes, including binding life events with context to form episodic memories, guiding context-relevant behavior, and recognizing objects within scenes and contexts., (© 2024 Wiley Periodicals LLC.)

Serotonin (5-HT) has long been involved in response to stress and its effect may be, in part, mediated by 5-HT1a and 5-HT7 receptor subtypes in different brain structures. Both pre- and post-synaptic activation of 5-HT1a receptor, respectively, in the rat median raphe nucleus (MnRN) and hippocampus, lead to adaptation to acute inescapable stressors such as restraint and forced swim. 5-HT7 receptor (5HT7r), a stimulatory G-protein coupled receptor, has also been investigated as a possible candidate for mediating stress response. In the MnRN, activation of 5-HT7r has antidepressant effects, while in the hippocampus, 5HT7r mRNA expression is increased after exposure to restraint stress, but the functional significance of these receptors remains to be determined. Therefore, the aim of this study was to investigate whether blockade of hippocampal 5HT7r would prevent and/or attenuate the behavioral effects of stress. Male adult Wistar rats with bilateral cannulas aimed at the dorsal hippocampus were restrained for 2 h and tested in the elevated plus maze (EPM) 24 h later. SB 258741 (3 nmoles/0.5 μL/side; selective 5HT7r antagonist) was administered bilateraly into the hippocampus according to the experimental protocol: immediately before or after stress, or 24 h after it (immediately before the test). In a second experiment, rats were exposed to 15 min. of forced swim, and tested 24 h later. Intra-hippocampal treatment was performed as described for the restraint stress protocol. We found that blockade of hippocampal 5-HT7r immediately after, but not before, the exposure to restraint or forced swim attenuated stress-induced behavioral changes. Similar results were obtained when SB was administered before the test in previously stressed rats. Our data suggest that activation of hippocampal 5-HT7r is crucial for the consolidation and retrieval of aversive stimulus-related memories, such as those caused by a stressful experience, probably through mechanisms involving stress-induced changes in 5-HT7r expression., (© 2024 Wiley Periodicals LLC.)

Massively multidimensional diffusion magnetic resonance imaging combines tensor-valued encoding, oscillating gradients, and diffusion-relaxation correlation to provide multicomponent subvoxel parameters depicting some tissue microstructural features. This method was successfully implemented ex vivo in microimaging systems and clinical conditions with tensor-valued gradient waveform of variable duration giving access to a narrow diffusion frequency (ω) range. We demonstrate here its preclinical in vivo implementation with a protocol of 389 contrast images probing a wide diffusion frequency range of 18 to 92 Hz at b-values up to 2.1 ms/μm 2 enabled by the use of modulated gradient waveforms and combined with multislice high-resolution and low-distortion echo planar imaging acquisition with segmented and full reversed phase-encode acquisition. This framework allows the identification of diffusion ω-dependence in the rat cerebellum and olfactory bulb gray matter (GM), and the parameter distributions are shown to resolve two water pools in the cerebellum GM with different diffusion coefficients, shapes, ω-dependence, relaxation rates, and spatial repartition whose attribution to specific microstructure could modify the current understanding of the origin of restriction in GM., (© 2024 The Author(s). NMR in Biomedicine published by John Wiley & Sons Ltd.)

Subarachnoid hemorrhage (SAH) is a specific type of stroke. Dihydroquercetin (DHQ), a flavonoid, is known for its various pharmacological properties. This study aimed to explore the roles and mechanisms of DHQ in influencing the progression of SAH. A rat SAH model was established using the endovascular perforation technique. Following SAH induction, DHQ was administered orally 1 h later. Assessments included SAH scores, neurological function, brain swelling, blood-brain barrier (BBB) integrity, neuronal damage, apoptosis levels, inflammation, and indicators of ferroptosis using various treatments. The HT22 cells were exposed to hemin to simulate SAH-like conditions under in vitro settings. Cell counting kit-8 assays, flow cytometry, enzyme?linked immunosorbent assay, BODIPY 581/591 C11 staining, western blot analysis, and biochemical kits were employed to evaluate the potential effects of DHQ. Moreover, the mechanisms responsible for the protective effect of DHQ were examined by western blot analysis. The in vivo findings revealed that DHQ mitigated neurological impairments, brain swelling, BBB disruption, and neuronal injury at 24 h post-SAH. DHQ also reduced neuronal degeneration, inflammation, and ferroptosis following SAH. The in vitro findings revealed that DHQ enhanced cell survival and reduced ferroptosis at 24 h following hemin exposure. Mechanistically, DHQ activated phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/protein kinase B (AKT)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling in SAH rats and hemin-treated HT22 cells to exert neuroprotective effects. In conclusion, this study reveals that DHQ can effectively decrease BBB permeability, brain edema, neurological dysfunctions, and ferroptosis post-SAH by activating the PI3K/AKT/Nrf2/HO-1 pathway., (© 2024 Wiley Periodicals LLC.)