Effects of moderate intensity endurance training and high-intensity interval training on the reproductive parameters of wistar rats overfed in infancy.

Studies indicate that rapid weight gain at critical development stages, such as the lactation period, is associated with the development of obesity, cardiovascular diseases, and diabetes in the long term. In addition to metabolic changes during adulthood, overweight/obesity may influence reproductive function. Human and animal studies suggest that lifestyle changes through exercise and/or controlled diet result in improved semen quality in obese individuals. However, the relationship between exercise volume/intensity and reproductive capacity effects remains inconclusive. The present study aimed to evaluate the effects of moderate intensity endurance training and high-intensity interval training (HIIT) on the reproductive parameters of lactating overfed male Wistar rats. Postnatal overfeeding was induced by applying the litter size reduction method. Forty males Wistar rats were used, divided into four groups: one with control litters (CLs) (10 animals/litter-sedentary) and three with small litters (SLs) (4 animals/litter), divided into sedentary, moderate endurance training, and HIIT. Morphologic, metabolic, and reproductive variables were analyzed. SL sedentary group showed increased body weight, adiposity, and decreased relative weight of the seminal vesicle, prostate, and epididymis as well as changes in the insulin tolerance and oral glucose tolerance tests glycemic tests compared to CL sedentary group. Endurance and HIIT protocols were efficient in improving the glycemic metabolism, central fat accumulation of trained groups and did not affect reproductive parameters. Endurance and HIIT protocols proved to be effective in reversing these metabolic changes without impairing the evaluated reproductive parameters.

Antimicrobial potential, phytochemical profile, cytotoxic and genotoxic screening of Sedum praealtum A. DC. (balsam).

BACKGROUND: Sedum praealtum has been used for a long time in traditional medicine as an analgesic and anti-inflammatory agent. Its beneficial effects have been known since ancient times, when Latinos used it to treat sore and swollen eyes. This research evaluated the antimicrobial potential, the cytotoxic and genotoxic effects, and some chromatographic profiles of the hydroethanolic extract of leaves, stems and roots of S. praealtum. METHODS: The antimicrobial activities were carried out by broth microdilution and agar diffusion. In vitro cytotoxicity was evaluated by cell cultures of Aedes albopictus and the selectivity index (SI) was estimated: SI=CI50/MIC. Genotoxic and systemic toxic effects of S. praealtum leaves were analyzed by micronucleus assay in mice bone marrow. Chromatographic profiles and mass spectra were investigated by GC-MS. RESULTS: Gram-positive (B. subtilis, B. cereus, M. luteus, E. faecalis and S. aureus) and gram-negative (E. coli, E. aerogenes, S. marcescens, P. aeruginosa, P. mirabilis and S. typhimurium) bacteria exhibited MICs ranging from 12.5-50 and 0-50 mg/ml, respectively. Sedum praealtum showed no efficacy against M. tuberculosis and M. bovis. Cytotoxicity (CI50) of S. praealtum was 4.22 and 5.96 mg/ml for leaves and stems, respectively, while its roots showed no cytotoxicity. Micronucleated polychromatic erythrocytes (MNPCEs) analyzes showed no differences between treatment doses (0.5-2 g/kg) and negative control (NaCl), but the PCE/NCE ratio (polychromatic erythrocyte/normochromatic erythrocyte) showed significant differences. Phytochemical screening identified thirteen compounds in the leaves, stems and roots of S. praealtum potentially associated with their biological activities. CONCLUSIONS: This research comprises a first scientific study on genotoxicity, cytotoxicity and antimicrobial effects of S. praealtum (Balsam), and it provides an initial theoretical foundation for its comprehensive use. Results showed antibacterial action of S. praealtum against gram-positive bacteria and some gram-negative species (depending on the plant anatomical part), but ineffective antimycobacterial action. However, S. praealtum leaves and stems display potential cytotoxicity, contributing to the SI < 1 values. In addition, S. praealtum leaves exhibit no clastogenic and/or aneugenic effects, but it has systemic toxicity dose-independent.

Digital radiography as an alternative method in the evaluation of bone density in uremic rats / Radiografia digital como método alternativo na avaliação da densidade óssea em ratos urêmicos

ABSTRACT Introduction: Digital radiography (DRx) may provide a suitable alternative to investigate mineral and bone disorder (MBD) and loss of bone density (BD) in rodent models of chronic kidney disease (CKD). The objective of this study was to use DRx to evaluate BD in CKD rats, and to evaluate the correlation between DRx findings and serum MBD markers and bone histomorphometry. Methods: Uremia was induced by feeding Wistar rats an adenine-enriched diet (0.75% for 4 weeks/0.10% for 3 weeks); outcomes were compared to a control group at experimental weeks 3, 4, and 7. The following biochemical markers were measured: creatinine clearance (CrC), phosphate (P), calcium (Ca), fractional excretion of P (FeP), alkaline phosphatase (ALP), fibroblast growth factor-23 (FGF-23), and parathyroid hormone (PTH). DRx imaging was performed and histomorphometry analysis was conducted using the left femur. Results: As expected, at week 7, uremic rats presented with reduced CrC and higher levels of P, FeP, and ALP compared to controls. DRx confirmed the lower BD in uremic animals (0.57±0.07 vs. 0.68 ± 0.06 a.u.; p = 0.016) compared to controls at the end of week 7, when MBD was more prominent. A severe form of high-turnover bone disease accompanied these biochemical changes. BD measured on DRx correlated to P (r=-0.81; p = 0.002), ALP (r = -0.69, p = 0.01), PTH (r = -0.83, p = 0.01), OS/BS (r = -0.70; p = 0.02), and ObS/BS (r = -0.70; p = 0.02). Conclusion: BD quantified by DRx was associated with the typical complications of MBD in CKD and showed to be viable in the evaluation of bone alterations in CKD.

RESUMO Introdução: A radiografia digital (RxD) pode representar uma alternativa adequada para investigar o distúrbio mineral e ósseo (DMO) e a perda de densidade óssea (DO) em modelos de roedores da doença renal crônica (DRC). O objetivo deste estudo foi utilizar a RxD para avaliar a DO em ratos com DRC, e avaliar a correlação entre os achados da RxD e marcadores séricos de DMO e histomorfometria óssea. Métodos: A uremia foi induzida pela alimentação de ratos Wistar com dieta enriquecida com adenina (0,75% por 4 semanas/0,10% por 3 semanas); os resultados foram comparados com um grupo controle nas semanas experimentais 3, 4 e 7. Os seguintes marcadores bioquímicos foram medidos: clearance de creatinina (CCr), fosfato (P), cálcio (Ca), fração excretada de P (FeP), fosfatase alcalina (ALP), fator de crescimento de fibroblastos-23 (FGF-23) e paratormônio (PTH). A imagem da RxD foi obtida e a análise histomorfométrica foi realizada com o fêmur esquerdo. Resultados: como esperado, na semana 7, os ratos urêmicos apresentaram redução do CCr e níveis mais altos de P, FeP e ALP em comparação aos controles. A RxD confirmou a menor DO em animais urêmicos (0,57 ± 0,07 vs. 0,68 ± 0,06 u.a.; p = 0,016) em comparação aos controles no final da semana 7, quando a DMO foi mais proeminente. Uma forma grave de doença óssea de alta renovação celular acompanhou essas mudanças bioquímicas. A DO, medida na RxD foi correlacionada a P (r = -0,81; p = 0,002), ALP (r = -0,69, p = 0,01), PTH (r = -0,83, p = 0,01), OS/BS (r = -0,70 p = 0,02) e Ob.S/BS (r = -0,70; p = 0,02). Conclusão: A DO quantificada por RxD esteve associada às complicações típicas da DMO na DRC e mostrou-se viável na avaliação de alterações ósseas na DRC.

A paternal hypercaloric diet affects the metabolism and fertility of F1 and F2 Wistar rat generations.

Increased fat and carbohydrate intakes based on the Western diet are important lifestyle modifications that lead to hypercaloric inputs, obesity, and male fertility negative effects. Epigenetic transmission may also predispose descended generations to chronic diseases, such as obesity, type 2 diabetes, behavioral, and reproductive disorders. The present study sought to evaluate the influence of a high-fat-high-sugar (HFHS) diet supplied to Wistar rats from 25 to 90 days of life on reproductive and metabolic parameters in male generations F0, F1, and F2. The standard group received the normocaloric - Nuvilab Quimtia® -3.86 kcal/kg. The hypercaloric diet (HD) group received the HFHS diet - PragSoluções® -4.77 kcal/kg. Body weight, adiposity, F1 and F2 prepubertal age evaluations, oral glucose tolerance test, insulin tolerance test, organ weights, sperm count and morphology assessments, and histometric testicular analyses were performed. The HFHS diet promoted dyslipidemia, higher adiposity, lower relative organ weights, and higher mean kidney weight, decreased mean testicle and parenchyma weights and lower height of seminiferous epithelium (HE) for the F0 generation. F1 and F2 offspring of HD group displayed early preprepubertal development, although did not alter the metabolic parameters. Decreased HE and tubular testicular compartment volumetric density and increased intertubular testicular compartment volumetric density and volume in the F1 generation of HD group were observed. Alterations in histometry of intertubular testicular compartment were also noted. It is concluded that the HFHS experimental model altered only paternal metabolic parameters. However, reproductive parameters of the three generations were affected.

Digital radiography as an alternative method in the evaluation of bone density in uremic rats.

INTRODUCTION: Digital radiography (DRx) may provide a suitable alternative to investigate mineral and bone disorder (MBD) and loss of bone density (BD) in rodent models of chronic kidney disease (CKD). The objective of this study was to use DRx to evaluate BD in CKD rats, and to evaluate the correlation between DRx findings and serum MBD markers and bone histomorphometry. METHODS: Uremia was induced by feeding Wistar rats an adenine-enriched diet (0.75% for 4 weeks/0.10% for 3 weeks); outcomes were compared to a control group at experimental weeks 3, 4, and 7. The following biochemical markers were measured: creatinine clearance (CrC), phosphate (P), calcium (Ca), fractional excretion of P (FeP), alkaline phosphatase (ALP), fibroblast growth factor-23 (FGF-23), and parathyroid hormone (PTH). DRx imaging was performed and histomorphometry analysis was conducted using the left femur. RESULTS: As expected, at week 7, uremic rats presented with reduced CrC and higher levels of P, FeP, and ALP compared to controls. DRx confirmed the lower BD in uremic animals (0.57±0.07 vs. 0.68 ± 0.06 a.u.; p = 0.016) compared to controls at the end of week 7, when MBD was more prominent. A severe form of high-turnover bone disease accompanied these biochemical changes. BD measured on DRx correlated to P (r=-0.81; p = 0.002), ALP (r = -0.69, p = 0.01), PTH (r = -0.83, p = 0.01), OS/BS (r = -0.70; p = 0.02), and ObS/BS (r = -0.70; p = 0.02). CONCLUSION: BD quantified by DRx was associated with the typical complications of MBD in CKD and showed to be viable in the evaluation of bone alterations in CKD.

Effects of moderate intensity endurance training vs. high intensity interval training on weight gain, cardiorespiratory capacity, and metabolic profile in postnatal overfed rats.

BACKGROUND: Obesity is associated with several comorbidities, such as cardiovascular disease and type 2 diabetes mellitus, and may have its origin in early life stages, such as in the lactation period, through metabolic programming. Physical activity aids in decreasing the chances of developing cardiovascular and metabolic diseases, even with small weight losses and, in children, can play an essential role in preventing weight gain and other health problems. The present study aimed to evaluate the effects of moderate intensity endurance training and high intensity interval training (HIIT) protocols on obesity-related parameters and cardiorespiratory capacity in overfed Wistar rats throughout the breastfeeding period. METHODS: Two days after birth, forty male and female Wistar rats were clustered into two groups: Control Litter Group (CL; ten animals/litter) and Reduced Litter Group (RL; four animals/litter). At weaning, RL animals were distributed randomly into three experimental groups: sedentary, moderate intensity endurance training and HIIT, while CL animals were clustered into a sedentary group. RESULTS: RL male and female body weight, before weaning, was significantly higher when compared with CL animals. This difference was maintained between CLSed and RLSed groups after weaning during all assessed periods. Adiposity was significantly higher in RLSed males when compared to CLSed males, and alterations in glycaemic metabolism were also observed. Endurance and HIIT protocols were efficient in improving maximal cardiorespiratory capacity, as well as concerning the glycemic metabolism and central fat accumulation of males and females submitted to childhood overfeeding by the litter reduction method. CONCLUSIONS: Both moderate endurance training and HIIT protocols included in early life were efficient in reverting or preventing certain metabolic alterations as a consequence of overfeeding during breastfeeding in male and female Wistar rats.

Avaliação do efeito da quercetina em ratos Wistar com síndrome metabólica / Evaluation of the effect of quercetin in Wistar rats with metabolic syndrome

Introdução: O tratamento da síndrome metabólica (SM) é um desafio, uma vez que terapias não medicamentosas são de difícil implementação e o tratamento farmacológico ideal não está totalmente estabelecido. Objetivo: Avaliar o efeito da quercetina na pressão arterial (PA), dislipidemia e acúmulo de gordura visceral em modelo experimental de SM induzida por dieta hiperlipídica. Material e Métodos: Ratos Wistar receberam ração hiperlipídica a partir da quarta semana de vida, por 20 semanas. O grupo tratado recebeu quercetina a partir da oitava semana de vida. Avaliou-se semanalmente o peso corporal e a PA de cauda por pletismografia. Ao final do experimento foram realizados testes de perfil glicêmico e lipídico. Resultados:A administração de dieta hiperlipídica se associou ao desenvolvimento de SM, caracterizada por acúmulo central de gordura, hipertensão arterial, hiperglicemia e hipertrigliceridemia. A quercetina não apresentou eficácia no tratamento das comorbidades que compõem a SM. Conclusão: A administração crônica diária da quercetina em modelo experimental de SM induzida por dieta hiperlipídica não alterou de forma significante o perfil nutricional, metabólico e pressórico dos animais.

Introduction: The treatment of the metabolic syndrome (MetS) is a challenge, since nonpharmacologic therapies are difficult to implement and the ideal pharmacologic treatment has not been completely established. Aim: To evaluate the effect of quercetin in blood pressure (BP), dyslipidemia, visceral fat accumulation, in an experimental model of MetS induced by a hyperlipidic diet. Material and Methods: Wistar rats received high fat diet feed from the fourth week of life for 20 weeks. The treatment group received quercetin from the eighth week of life. Body weight and tail BP through pletysmography were evaluated weekly. At the end of the experiment, tests of glucose and lipid profile. Results: The administration of a high fat diet was associated to the development of MetS, characterized by an accumulation of central fat, arterial hypertension, hyperglycemia, and hypertriglyceridemia. Quercetin was not effective in the treatment of comorbidities associated with MetS. Conclusion: Chronic daily administration of quercetin in an experimental model of MetS induced by a hyperlipidic diet did not significantly alter the nutritional, metabolic, and pressure profile of the animals.

Acute and sub chronic toxicity study of aqueous extract from the leaves and branches of Campomanesia velutina (Cambess) O. Berg.

ETHNOPHARMACOLOGICAL RELEVANCE: Campomanesia velutina leaves and branches infusions are used in Brazilian folk medicine to treat diarrhea and to ameliorate intestinal cramps, respectively. AIM OF THE STUDY: Carry out the acute and sub chronic pre-clinical evaluation and thus assess the safety and toxicological potential of the specie. MATERIALS AND METHODS: In vivo toxicity was evaluated by acute and sub chronic toxicity assays conducted according to the guidelines of the Brazilian Agency of National Health Surveillance (Agência Nacional de Vigilância Sanitária - ANVISA). For acute toxicity evaluation, a single dose of aqueous extracts from the leaves (AEL) and branches (AEB) of Campomanesia velutina were orally administered to mice at doses of 300, 600 and 1200mg/kg. Then, the animals were observed for 14 days. In the sub chronic study, the extracts were orally administered to mice for 14 days at doses of 300, 600 and 1200mg/kg. To assess the toxicological effects, animals were closely observed on general behavior, clinical signs of toxicity, body weight, food and water intake. At the end of the experiment, it was performed biochemical and hematological evaluations, as well as histopathological analysis from the following organs: brain, heart, lungs, liver, stomach, small intestine (section) and left kidney. Preliminary phytochemical analysis was performed using thin layer chromatography (TLC) and colorimetric pharmacognostic tests. RESULTS: In oral acute assay, treatment with AEB at the major dose (1200mg/kg) caused diarrhea, abdominal cramps and tremors in females. These effects were reversed at 4th hour. Normochromic normocytic anemia was observed in males treated with AEL 300mg/kg and AEB 600 and 1200mg/kg as well as in females treated with AEB 300 and 1200mg/kg. The kidney of all treated animals showed moderate inflammation and a few hemorrhagic points. In sub chronic assay, treatment with AEL 600mg/kg, AEL 1200mg/kg and AEB 1200mg/kg caused hyper excitability in females that was not reversed. Treatments also had impact on weight gain and the relative weight of males' brain was increased on group treated with AEL 300mg/kg, AEB 300 and AEB 1200mg/kg. Although changes in hematological parameters were not observed, serum creatinine levels were significantly higher in males treated with AEB 300mg/kg. Besides, the heart of all treated animals showed intense hyperemia. Preliminary phytochemical analysis revealed the presence of flavonoids, tannins and phenolic compounds. CONCLUSIONS: Toxicity signs were mainly observed after treatment with AEL and AEB at the two highest tested doses (600 and 1200mg/kg), suggesting that the extracts are relatively safe at its effective dose (300mg/kg). However, alterations on hematological and biochemical parameters and on the kidney and heart of the animals were not closely related with the dose, implying caution on its use.

Effect of mesenchymal stem cells and mouse embryonic fibroblasts on the development of preimplantation mouse embryos.

Despite advances in assisted reproduction techniques, the poor quality and failures in embryo in vitro development remain as drawbacks resulting in low pregnancy rate. Mouse embryonic fibroblasts (MEFs) have been widely used to support embryonic stem cells. Mesenchymal cells (MSCs) have also been shown to release bioactive factors. In the present study, we have evaluated the ability of MSCs and MEFs to support early development of mouse embryos. The embryos were cultivated alone or in coculture with inactivated MSC or MEF for 4 d. After 4 d in culture, the percentage of blastocyst formation in coculture with MSC (91.7 ± 4.3%) or MEF (95.1 ± 3.3%) was higher than in the control group (72.2 ± 9.0%). We did not observe any difference in proliferation or apoptosis. However, the blastocysts cocultured with MSC or MEF presented a significantly higher number of cells within the inner cell mass per embryo when compared to the controls. The MSC and MEF groups presented also a higher cell number and diameter when compared to the control (CTRL). In summary, our data indicate that coculture with MSC or MEF improves early embryonic development and quality in vitro.

Ausência de toxicidade reprodutiva e sistêmica em ratos wistar tratados com extrato seco de gingko biloba durante o período de desmame à puberdade / Absence of reproductive and sistemic toxicity in rats treated with Gingko biloba extract from weaning to puberty

Introdução: O Gingko biloba (EGb) é um fitoterápico usado há séculos, porém com poucos estudos referentes a seus efeitos sobre o período pós-natal. Estudos dessa natureza vêm sendo preconizados pela Agência Europeia de Medicina, visto que muitos órgãos completam seu desenvolvimento nesse período, inclusive o sistema reprodutor. Objetivo: Avaliar o efeito do extrato seco de EGb sobre o desenvolvimento do sistema reprodutor de ratos, tratados desde o desmame até o fim da puberdade. Métodos. Ratos Wistar foram tratados com 25mg/kg/massa corporal (EGb 25); 50 mg/kg (EGb 50) e 100 mg/kg (EGb 100). Controle (C ­ 0,1ml água destilada), por gavage dos 25 aos 45 dias de vida pós-natal. Variáveis observadas: indícios clínicos de toxicidade sistêmica, peso corporal, descida dos testículos, evolução da morfologia da glande, peso de rins, baço e fígado e dos órgãos do sistema reprodutor. Hematimetria, Concentração de hemoglobina. Concentração de espermatozoides na secreção epididimária. Resultados: Não foram encontradas diferenças significativas em quaisquer das variáveis. Conclusão: A exposição ao extrato seco de EGb durante o período pré-puberal e puberal em ratos Wistar não altera o desenvolvimento do sistema reprodutor masculino.

Introduction: Gingko biloba extract (EGb) is a phytotherapic that has been used for centuries but there is no studies concerning their effects during the postnatal period. This kind of research had been suggested by the European Medicine Agency since there are organs that complete their development in this period, including reproductive organs. Purpose: To evaluate the effect of EGb dry extract upon the rat reproductive system from weaning to 45 postnatal days. Methods: Wistar rats were treated with 25mg/kg/body weight (EGb 25); 50 mg/kg (EGb 50) and 100 mg/kg (EGb 100). Control (C 0,1ml distilled water). Variables: clinical signs of systemic toxicity, body weight, testicles descent, evolution of glans morphology, kidneys, liver, spleen and reproductive organs weights. Hematimetry. Haemoglobin concentration. Sperm concentration in the epidydimal secretion. Results: No significant differences were observed in none of the observed variables. Conclusion: The EGb dry extract exposition to prepuberal and puberal rats do not alter the reproductive system development.

Toxicological evaluation of the flavonoid rutin on the reproductive system of wistar rats / Avaliação toxicológica do flavanoide rutina no sistema reprodutor de ratos wistar

Introduction: Rutin, a flavonoid commonly found in nature, has anti-mitotic, vasoprotective, and antihyperlipidemic activity. When hydrolyzed as quercetin, it promotes inhibition of spermatozoa motility, alterations in the prostate, and in the levels of testosterone and dihydrotestosterone. Objective: This study aimed to evaluate the toxicity of rutin in Wistar rats. Methods: Animals were divided into Control (1 ml of distilled water), Treated I, II and III, respectively receiving 5, 10 and 20 mg/kg/day of rutin for seven consecutive days. When euthanasia was performed after 10, 42 and 60 days into the experiment, a laparotomy was performed and the testicles, prostate, seminal vesicles, epididymis, epididymal spermatozoa to be counted, as well as the liver, spleen and kidneys were removed. Hematological and biochemical tests were performed. Results: Hepatomegaly was observed and in the reproductive system, the weight of the epididymis was reduced, not affecting any other organ examined. Conclusion: Except by the reduction of the weight of the epididymis, which is reversible at 42 days of completion of treatment, no suggestive data of the toxicity of rutin on the reproductive system of adult rats were found.

Efeito do extrato de ginkgobiloba (EGb) sobre a toxicidade sistêmica e órgãos do sistema reprodutor masculino de ratos Wistar adultos / Effect of ginkgobiloba extract (EGb) on systemic toxicity and organs of the male reproductive system of adult Wistar rats

O Extrato de Ginkgobiloba (EGb) é um dos fitoterápicos mais consumidos no mundo. Entretanto ainda há escassez de ensaios toxicológicos em animais e o risco à exposição humana principalmente pelos compostos alquilfenóis, representados pelos ácidos ginkgólicos, que podem causar quadros alérgicos e serem compostos mutagênicos e carcinogênicos. O presente trabalho teve o objetivo de avaliar a toxicidade sistêmica do EGb. Oitenta ratos Wistar de três meses de idade foram tratados com água destilada (Grupo Controle) e extrato aquoso de Ginkgobilobanas seguintes doses: 3,5 (EGb 3,5); 7,0 (EGb 7,0) e 14,0mg/kg (EGb 14,0) uma vez ao dia, por 56 dias consecutivos. Foram avaliados semanalmente, o peso dos animais (g) e a estimativa de consumo diário de ração (g). Indícios de sinais de toxicidade sistêmica como perda de peso, piloereção, diarreia, cromodacriorreia, estereotipias, alterações da atividade locomotora e comportamentais e mortes também foram monitorados. Após anestesia, o sangue dos animais foi coletado para avaliação de hemograma completo e dosagem bioquímica de ureia, creatinina e alanina aminotransferase (ALT). Após a eutanásia, os animais foram submetidos à necropsia e os testículos esquerdo e direito, epidídimo esquerdo, vesícula seminal repleta, próstata ventral, rins esquerdo e direito, fígado e baço foram removidos e pesados em balança de precisão. Durante todo o procedimento experimental não foram observados nos animais sinais clínicos de toxicidade sistêmica e mortes. Houve diferenças estatísticas da estimativa de consumo de ração na sexta semana e oitava semanas de avaliação, embora sem diferença no peso corporal. Não houve diferença no peso dos órgãos e na análise bioquímica sérica. Na avaliação hematológica dos animais, houve diferença estatística significativa na hemoglobinometria em que o grupo EGb 14,0 apresentou-se estatisticamente superior ao grupo EGb 3,5.A concentração de hemoglobina globular média também apresentou diferença estatística significativa, em que o EGb 3,5 apresentou médias inferiores aos grupos EGb 7,0 e EGb 14,0 e o grupo controle apresentou média inferior ao grupo EGb 14,0. Sugere-se que o EGb no presente trabalho, e com as doses utilizadas, não causou toxicidade sistêmica e nem provocou alterações em órgãos de ratos Wistar.

The Ginkgobiloba Extract (EGb) is one of the most commonly consumed herbal in the world. However there are still few toxicity tests on animals and the risk of human exposure mainly by alkyl compounds, represented by acids, which can cause allergies and are mutagenic and carcinogenic compounds. This study had the objective of evaluate the systemic toxicity of EGb. Eighty Wistar rats, three months of age were treated with distilled water (Control Group) and aqueous extract Ginkgobilobanas following doses: 3.5 (EGb 3.5); 7.0 (EGb 7.0) and 14,0mg / kg (14.0 EGb) once a day for consecutive 56 days. Were evaluated weekly animal weight (g) and the estimated daily intake (g). Evidence of systemic signs of toxicity such as weight loss, piloerection, diarrhea, stereotypies and behavioral changes in motor activity and deaths were also monitored. After anesthesia, the animals were collected for evaluation of complete blood count and biochemical analysis of urea, creatinine and alanine aminotransferase (ALT). After euthanasia, the animals were autopsied and the left and right testis, left epididymis, seminal vesicle filled, ventral prostate, left and right kidneys, liver and spleen were removed and weighed on a precision scale. Throughout the experimental procedure were not observed in animals clinical signs of systemic toxicity and deaths. Were no statistical differences in the estimate of feed intake in the sixth week and eighth week evaluation, although no difference in body weight. There were no differences in organ weight and serum biochemical analysis. Hematological evaluation of the animals, there was a statistically significant difference in Hemoglobinometry where 14.0 EGb group was statistically higher than the EGb group 3,5. A mean corpuscular hemoglobin concentration also showed a statistically significant difference in the EGb 3 5 showed an average lower than 7.0 and EGb groups EGb 14.0 and the control group showed less than 14.0 EGb group. It is suggested that EGb in this work, and the doses used, did not cause systemic toxicity nor caused changes in organs of Wistar rats.

Molecular imaging, biodistribution and efficacy of mesenchymal bone marrow cell therapy in a mouse model of Chagas disease.

Chagasic cardiomyopathy, resulting from infection with the parasite Trypanosoma cruzi, was discovered more than a century ago and remains an incurable disease. Due to the unique properties of mesenchymal stem cells (MSC) we hypothesized that these cells could have therapeutic potential for chagasic cardiomyopathy. Recently, our group pioneered use of nanoparticle-labeled MSC to correlate migration with its effect in an acute Chagas disease model. We expanded our investigation into a chronic model and performed more comprehensive assays. Infected mice were treated with nanoparticle-labeled MSC and their migration was correlated with alterations in heart morphology, metalloproteinase activity, and expression of several proteins. The vast majority of labeled MSC migrated to liver, lungs and spleen whereas a small number of cells migrated to chagasic hearts. Magnetic resonance imaging demonstrated that MSC therapy reduced heart dilatation. Additionally metalloproteinase activity was higher in heart and other organs of infected mice. Protein expression analyses revealed that connexin 43, laminin γ1, IL-10 and INF-γ were affected by the disease and recovered after cell therapy. Interestingly, MSC therapy led to upregulation of SDF-1 and c-kit in the hearts. The beneficial effect of MSC therapy in Chagas disease is likely due to an indirect action of the cells of the heart, rather than the incorporation of large numbers of stem cells into working myocardium.

The effect of the Ginkgo biloba extract in the expression of Bax, Bcl-2 and bone mineral content of Wistar rats with glucocorticoid-induced osteoporosis.

OBJECTIVE: Evaluate the effects of the extract of Ginkgo biloba (EGb) in the glucocorticoid-induced-osteoporosis through the Bax and Bcl-2 expressions by osteoblast cells, the x-ray and bone density of the tibia. METHOD: Rats were divided into five groups: osteoporosis; EGb1 (28 mg/kg); EGb2 (56 mg/kg); alendronate (0.2 mg/animal) and control. The treatments were conducted for 20 (n = 30) and 30 days (n = 30). The Bax and Bcl-2 expressions were evaluated in osteoblasts of the mandibular alveolar bone. The tibias were radiographed to evaluate the X-ray and bone density. The control group was compared with the osteoporosis' (Student's t-test/Mann-Whitney). The other groups were analyzed by analysis of variance test followed by Dunnett/Dunnett T3 (p < 0.05). RESULTS: When compared the osteoporosis to the control group (p <0.05): Bax and x-ray density increased; Bcl-2 and the bone density reduced. When compared with the osteoporosis group (p < 0.05), alendronate (30 days), EGb1 and EGb2 (20/30 days) increased the Bcl-2 expression; EGb2 and alendronate (20 days) EGb1 and EGb2 (30 days) reduced the Bax expression; and EGb1 and EGb2 (20/30 days) reduced the X-ray density. CONCLUSIONS: The EGb improved the Bcl-2 and reduced the Bax expression by osteoblasts in the mandibular alveolar bone and recovered the mineral content in the tibia of rats with glucocorticoid-induced-osteoporosis.

Dissociação da resposta anti-hipertensiva e metabólica à losartana e espironolactona em ratos com síndrome metabólica experimental / Dissociation of antihypertensive and metabolic response to losartan and spironolactone in experimental rats with metabolic sindrome

INTRODUÇÃO: O tratamento da hipertensão arterial (HA) em indivíduos com síndrome metabólica (SM) é um desafio, uma vez que terapias não medicamentosas são de difícil implementação e o tratamento farmacológico ideal não está totalmente estabelecido. OBJETIVO: Avaliar o bloqueio do sistema renina angiotensina aldosterona (SRAA) na pressão arterial (PA), na função e na morfologia renais em modelo experimental de SM induzida por dieta hiperlipídica. MÉTODOS: Ratos Wistar receberam ração hiperlipídica a partir da quarta semana de vida, por 20 semanas. Os grupos tratados receberam Losartana ou Espironolactona a partir da oitava semana de vida. Avaliou-se semanalmente o peso corporal e a PA de cauda por pletismografia. Ao final do experimento, foram realizados testes de tolerância oral à glicose, perfil lipídico, clearance de creatinina, medida direta da PA, análise morfométrica renal. RESULTADOS: A administração de dieta hiperlipídica se associou ao desenvolvimento de SM, caracterizada por acúmulo central de gordura, hipertensão arterial, hiperglicemia e hipertrigliceridemia. Nesse modelo não foram observadas alterações da histomorfometria renal. O bloqueio do receptor AT1 da angiotensina II (Ang II) preveniu o desenvolvimento da HA. O bloqueio mineralocorticoide não apresentou eficácia anti-hipertensiva, porém, associou-se à redução da gordura abdominal. CONCLUSÃO: A dissociação da resposta anti-hipertensiva aos bloqueios dos receptores da Ang II e mineralocorticoide indica a participação da Ang II na gênese da HA associada à obesidade. A redução da obesidade central com a Espironolactona sugere a presença de efeito adipogênico mineralocorticoide.

INTRODUCTION: The treatment of arterial hypertension (AH) in patients with metabolic syndrome (MS) is a challenge, since non drug therapies are difficult to implement and optimal pharmacological treatment is not fully established. OBJECTIVE: To assess the blockade of the rennin angiotensin aldosterone system (RAAS) in blood pressure (BP) in renal function and morphology in an experimental model of MS induced by high fat diet. METHODS: Wistar rats were fed on high fat diet from the fourth week of life, for 20 weeks. The groups received Losartan or Spironolactone from the eighth week of life. We weekly evaluated the body weight and BP by tail plethysmography. At the end of the experiment oral glucose tolerance, lipid profile, creatinine clearance tests, and the direct measurement of BP were performed. A morphometric kidney analysis was performed. RESULTS: The administration of high-fat diet was associated with the development of MS, characterized by central fat accumulation, hypertension, hyperglycemia and hypertriglyceridemia. In this model there were no changes in renal histomorphometry. The blockade of angiotensin II (Ang II) receptor AT1 prevented the development of hypertension. The mineralocorticoid blockage did not have antihypertensive efficacy but was associated with reduction of abdominal fat. CONCLUSION: The dissociation of the antihypertensive response to the blockades of Ang II receptors and mineralocorticoid indicates the involvement of Ang II in the pathogenesis of hypertension associated with obesity. Reduction of central obesity with Spironolactone suggests the presence of mineralocorticoid adipogenic effect.

[Effect of ipriflavone on Wistar rats and their litters]. / Efeito da ipriflavona sobre ratas Wistar e suas ninhadas.

PURPOSE: Evaluate the effects of ipriflavone during fetogenesis, since no studies have been conducted to assess its effect during this period. METHODS: 60 pregnant rats were divided randomly into four groups (n=15). G-control (1 mL of distilled water) and three groups treated intragastrically with ipriflavone from the 16th to the 20th post coitus (PC) day: G-300 (300 mg/kg), G-1,500 (1,500 mg/kg) and G-3,000 (3,000 mg/kg). The animals were weighed, anaesthetized intraperitoneally with xylazine and ketamine at doses of 180 mg/kg and 10 mg/kg, respectively, and sacrificed by total exsanguination on the 21st day. A complete blood count was performed and serum cholesterol, triglycerides, AST, ALT, urea, creatinine, and glucose were determined in pregnant rats. After laparotomy, the liver, kidneys, adrenals, spleen and ovaries were removed and weighed; fetuses and placentas were also weighed to obtain the average weight of the litters. Four fetuses (two males and two females) were chosen at random for the determination of the length and weight of brain, liver, kidneys and lungs. STATISTICAL ANALYSIS: ANOVA followed by Dunnett's test. For raw data without normal distribution and homoscedasticity, we used the Kruskal-Wallis test followed by the Mann-Whitney test. Proportions were analyzed by the χ² test (p<0.05). RESULTS: Triglyceride levels (mg/dL) were: Control-G (138.8±21.8), G-300 (211.2±63.9) G-1,500 (251.5±65.2) G-3,000 (217.7±49.6); p<0.05. The body weight of fetuses (g) was: G-Control (male 3.3±0.3; female 3.1±0.3), G-300 (male 3.4±0.2; female 3.1±0.4), G-1,500 (male 3.5±0.3; female 3.2±0.3), G-3,000 (male 3.4±0.5; female 3.1±0.4). CONCLUSION: Ipriflavone did not cause maternal toxicity, but increased triglyceride levels and reduced hematocrit at higher doses. The body and organ weights of the fetuses did not change with dam treatment. There were no external malformations or fetal deaths.

Efeito da ipriflavona sobre ratas Wistar e suas ninhadas / Effect of ipriflavone on Wistar rats and their litters

OBJETIVO: Avaliar os efeitos da ipriflavona durante a fetogênese, já que não foram encontrados estudos visando avaliar seu efeito durante este período. MÉTODOS: Foram utilizadas 60 ratas prenhes, distribuídas aleatoriamente em quatro grupos (n=15). G-controle (1 mL de água destilada) e três grupos tratados com ipriflavona, via intragástrica, do 16º ao 20º dia pós-coito (PC): G-300 (300 mg/kg), G-1500 (1.500 mg/kg) e G-3000 (3.000 mg/kg). Os animais foram pesados e sacrificados no 21º dia por exsanguinação total sob anestesia (xilazina (10 mg/kg) e quetamina (90 mg/kg) via intraperitoneal. Foi realizado hemograma completo e dosagens séricas de colesterol, triglicérides, AST, ALT, ureia, creatinina e glicose das ratas prenhes. Após laparotomia foram removidos e pesados fígado, rim, suprarrenais, baço e ovários; os fetos e placentas foram pesados obtendo-se o peso médio das ninhadas. Quatro fetos (dois machos e duas fêmeas) por mãe foram aleatoriamente designados para obter-se o comprimento e peso de cérebro, fígado, rins e pulmões. Para a análise estatística utilizou-se o teste ANOVA seguido do teste de Dunnet; para dados não homocedásticos e sem distribuição normal, foi usado o teste de Kruskal-Wallis, seguido de Mann-Whitney; as proporções foram analisadas pelo teste do χ² (p<0,05) RESULTADOS: Níveis de triglicérides (mg/dL): G-Controle (138,8±21,8); G-300 (211,2±63,9); G-1500 (251,5±65,2); G-3000 (217,7±49,6); p<0.05. Peso corporal dos fetos (g): G-Controle (machos 3,3±0,3; fêmeas 3,1±0,3); G-300 (machos 3,4±0,2; fêmeas 3,1±0,4); G-1500 (machos 3,5±0,3; fêmeas 3,2±0,3); G-3000 (machos 3,4±0,5; fêmeas 3,1±0,4). CONCLUSÃO: A ipriflavona não causou toxicidade materna, mas elevou níveis de triglicérides e reduziu o hematócrito em doses elevadas, o tamanho, peso corporal e de órgãos fetais não foram alterados. Não foram observadas malformações externas nem mortes fetais.

PURPOSE: Evaluate the effects of ipriflavone during fetogenesis, since no studies have been conducted to assess its effect during this period. METHODS: 60 pregnant rats were divided randomly into four groups (n=15). G-control (1 mL of distilled water) and three groups treated intragastrically with ipriflavone from the 16th to the 20th post coitus (PC) day: G-300 (300 mg/kg), G-1,500 (1,500 mg/kg) and G-3,000 (3,000 mg/kg). The animals were weighed, anaesthetized intraperitoneally with xylazine and ketamine at doses of 180 mg/kg and 10 mg/kg, respectively, and sacrificed by total exsanguination on the 21st day. A complete blood count was performed and serum cholesterol, triglycerides, AST, ALT, urea, creatinine, and glucose were determined in pregnant rats. After laparotomy, the liver, kidneys, adrenals, spleen and ovaries were removed and weighed; fetuses and placentas were also weighed to obtain the average weight of the litters. Four fetuses (two males and two females) were chosen at random for the determination of the length and weight of brain, liver, kidneys and lungs. Statistical analysis: ANOVA followed by Dunnett's test. For raw data without normal distribution and homoscedasticity, we used the Kruskal-Wallis test followed by the Mann-Whitney test. Proportions were analyzed by the χ² test (p<0.05). RESULTS: Triglyceride levels (mg/dL) were: Control-G (138.8±21.8), G-300 (211.2±63.9) G-1,500 (251.5±65.2) G-3,000 (217.7±49.6); p<0.05. The body weight of fetuses (g) was: G-Control (male 3.3±0.3; female 3.1±0.3), G-300 (male 3.4±0.2; female 3.1±0.4), G-1,500 (male 3.5±0.3; female 3.2±0.3), G-3,000 (male 3.4±0.5; female 3.1±0.4). CONCLUSION: Ipriflavone did not cause maternal toxicity, but increased triglyceride levels and reduced hematocrit at higher doses. The body and organ weights of the fetuses did not change with dam treatment. There were no external malformations or fetal deaths.

Efficacy of Morus nigra L. on reproduction in female Wistar rats.

Morus nigra L. is a plant employed as a substitute for the conventional hormonal replacement therapy. This work analyzes the estrogenic effect of M. nigra on the reproductive system and embryonic development of Wistar rats. Female rats were orally treated with M. nigra hydroalcoholic extract (MnHE) at the dose levels of 25, 50, 75, 350 and 700 mg/kg of body weight over 15 days, and continued through mating until the 14th day of gestation. Vaginal smears were performed daily and the body weight of the females was recorded at 5 days intervals. On day 15 of gestation, the females were killed and their kidneys, liver, spleen and ovaries were removed and weighed. The number of implants, resorptions, and live and dead fetuses were evaluated. Histological sections of ovaries, measurement of the height of the uterine epithelium and vaginal smears were performed to assess the estrogenic activity. The results showed that the administration of MnHE did not significantly alter the analyzed variables. Therefore, considering the experimental model used in this study, the data obtained indicate that M. nigra did not exhibit any estrogenic activity nor did exert a toxic effect on the female reproductive system and on the embryonic development of rats.

[Dissociation of antihypertensive and metabolic response to losartan and spironolactone in experimental rats with metabolic sindrome]. / Dissociação da resposta anti-hipertensiva e metabólica à losartana e espironolactona em ratos com síndrome metabólica experimental.

INTRODUCTION: The treatment of arterial hypertension (AH) in patients with metabolic syndrome (MS) is a challenge, since non drug therapies are difficult to implement and optimal pharmacological treatment is not fully established. OBJECTIVE: To assess the blockade of the rennin angiotensin aldosterone system (RAAS) in blood pressure (BP) in renal function and morphology in an experimental model of MS induced by high fat diet. METHODS: Wistar rats were fed on high fat diet from the fourth week of life, for 20 weeks. The groups received Losartan or Spironolactone from the eighth week of life. We weekly evaluated the body weight and BP by tail plethysmography. At the end of the experiment oral glucose tolerance, lipid profile, creatinine clearance tests, and the direct measurement of BP were performed. A morphometric kidney analysis was performed. RESULTS: The administration of high-fat diet was associated with the development of MS, characterized by central fat accumulation, hypertension, hyperglycemia and hypertriglyceridemia. In this model there were no changes in renal histomorphometry. The blockade of angiotensin II (Ang II) receptor AT1 prevented the development of hypertension. The mineralocorticoid blockage did not have antihypertensive efficacy but was associated with reduction of abdominal fat. CONCLUSION: The dissociation of the antihypertensive response to the blockades of Ang II receptors and mineralocorticoid indicates the involvement of Ang II in the pathogenesis of hypertension associated with obesity. Reduction of central obesity with Spironolactone suggests the presence of mineralocorticoid adipogenic effect.

Avaliação do potencial interceptivo da ipriflavona em ratas wist / Evaluation of the potential of interceptive ipriflavone in wistar rats

A ipriflavona possui efeito inibitório sobre o citocromo p450 e sobre a proliferação do DNA, interferindo na síntese de hormônios estereoidianos, e consequente interferência no transporte e desenvolvimento pré-implantacional e na implantação do blastocisto. Consumidos em altas doses pode acarretar abortamentos e malformações. Para verificar a embriotoxicidade da ipriflavona durante as fases de transporte tubário e implantação do blastocisto de ratas, ratas Wistar prenhes foram tratadas duas vezes ao dia com 1mL de suspensão aquosa de ipriflavona, nas doses de 0 (C), 300 (T1), 1500 (T2) e 3000 (T3) mg/kg/dose, durante os oito primeiros dias de prenhez com eutanásia no 14o dia. Indícios de toxicidade foram avaliados por análises hematimétricas, bioquímicas e comportamentais. Foram contados fetos vivos, mortos e reabsorvidos (inicial e tardiamente). Os animais não apresentaram sinais clínicos de toxicidade. Índice de implantação e perdas pré-embrionária sofreram interferência do uso da ipriflavona. Dados apontam para um efeito estrogênico da ipriflavona, observados na interferência da implantação do blastocisto de ratas Wistar.

Ipriflavone has inhibitory effect on the cytochrome p450 and the proliferation of DNA, interfering with hormone synthesis estereoidianos, and consequent interference with the transport and preimplantation development and implantation of the blastocyst. Consumed in high doses can cause miscarriages and malformations. To verify the embryotoxicity of ipriflavone during the phases of tubal transport and implantation of the blastocyst in rats, Wistar rats were treated twice daily with 1 mL of aqueous suspension of ipriflavone at doses of 0 (C), 300 (T1), 1500 (T2) and 3000 (T3) mg / kg / dose during the first eight days of pregnancy with euthanasia on day 14. Signs of toxicity were characterized by hematological, biochemical and behavioral. Live fetuses were counted, dead and resorbed (early and late). The animals showed no clinical signs of toxicity. Index pre-implantation embryo and losses suffered interference from the use of ipriflavone. Data point to an estrogenic effect of ipriflavone, due to interference in blastocyst implantation in Wistar rats.