Saccharomyces boulardii modulates oxidative stress and renin angiotensin system attenuating diabetes-induced liver injury in mice.

Type 1 diabetes (T1DM) is a chronic disease characterized by hyperglycemia due to a deficiency in endogenous insulin production, resulting from pancreatic beta cell death. Persistent hyperglycemia leads to enhanced oxidative stress and liver injury. Several studies have evaluated the anti-diabetic and protective effects of probiotic strains in animal models. In the present study, we investigated, through histopathological and biochemical analyses, the effects of eight weeks of administration of Saccharomyces boulardii (S. boulardii) yeast on the liver of streptozotocin (STZ) induced diabetic C57BL/6 mice. Our results demonstrated that S. boulardii attenuates hepatocytes hydropic degeneration and hepatic vessels congestion in STZ-induced diabetic mice. The treatment attenuated the oxidative stress in diabetic mice leading to a reduction of carbonylated protein concentration and increased activity of antioxidant enzymes superoxide dismutase and glutathione peroxidase, compared to untreated diabetic animals. The results also show the beneficial influence of S. boulardii in regulating the hepatic concentration of renin angiotensin system (RAS) peptides. Therefore, our results demonstrated that S. boulardii administration to STZ-induced diabetic mice reduces oxidative stress and normalizes the concentration of RAS peptides, supporting the hypothesis that this yeast may have a role as a potential adjunctive therapy to attenuate diabetes-induced liver injury.

Intense resistance training induces pronounced metabolic stress and impairs hypertrophic response in hind-limb muscles of rats.

Skeletal muscle hypertrophy is an exercise-induced adaptation, particularly in resistance training (RT) programs that use large volumes and low loads. However, evidence regarding the role of rest intervals on metabolic stress and muscular adaptations is inconclusive. Thus, we aimed to investigate the effects of a strenuous RT model (jump-training) on skeletal muscle adaptations and metabolic stress, considering the scarce information about RT models for rats. We hypothesized that jump-training induces metabolic stress and influences negatively the growth of soleus (SOL) and extensor digitorum longus (EDL) muscles of rats. Male Wistar rats (aged 60 days) were randomly assigned to non-trained or trained groups (n = 8/group). Trained rats performed jump-training during 5 days a week for 1, 3, or 5 weeks with 30 s of inter-set rest intervals. Forty-eight hours after the experimental period, rats were euthanized and blood samples immediately drawn to measure creatine kinase activity, lactate and corticosterone concentrations. Muscle weight-to-body weight ratio (MW/BW), cross-sectional area (CSA) and myosin heavy chain (MHC) isoform expression were determined. Higher lactate levels occurred after 20 min of training in weeks 1 and 3. Corticosterone levels were higher after 5 weeks of training. Jump-training had negative effects on hypertrophy of types-I and II muscle fibers after 5 weeks of training, as evidenced by decreased CSA and reduced muscle weight. Our results demonstrated that pronounced metabolic stress and impairment of muscle growth might take place when variables of exercise training are not appropriately manipulated. Lay summary Resistance training (RT) has been used to increase muscle mass. In this regard, training variables (intensity, volume, and frequency) must be strictly controlled in order to evoke substantial muscular fitness. This study shows that rats submitted to 5 weeks of intensive resistance jump-training - high intensity, large volume, and short rest intervals - present high levels of blood corticosterone associated with negative effects on hypertrophy of types-I and II muscle fibers.

90-kDa N-domain angiotensin I-converting enzyme (ACE): possible marker for hypertension in a renal transplant model.

INTRODUCTION: Hypertension is nearly universal in kidney transplant and several factors are associated with post transplant hypertension, including immunosuppressive medications and genetic predisposition. OBJECTIVE: The aims were to evaluate the effects of spontaneously hypertensive rats (SHR) kidney transplantation in Wistar rats and the possible transference of 80/90-kDa N-domain ACE. METHODS: To do so, the data from Wistar recipients of kidney from SHR were compared to data from transplanted Wistar submitted to CsA treatment and, to Wistar Sham. RESULTS AND DISCUSSION: Despite the unaltered blood pressure observed at early stages, 80/90-kDa ACE was found expressed in the urine of rats 7 and 15 days after transplantation, which was intense when rats became hypertensive 30 days post-surgery. CONCLUSION: Our data show that this enzyme is associated with the development of hypertension, and this marker appears in the urine before any substantial blood pressure alteration.

90-kDa N-domain angiotensin I-converting enzyme (ACE): possible marker for hypertension in a renal transplant model / Enzima conversora de angiotensina I (ECA) N-domínio 90-kDa: possível marcador para hipertensão em modelo de transplante renal

Abstract Introduction: Hypertension is nearly universal in kidney transplant and several factors are associated with post transplant hypertension, including immunosuppressive medications and genetic predisposition. Objective: The aims were to evaluate the effects of spontaneously hypertensive rats (SHR) kidney transplantation in Wistar rats and the possible transference of 80/90-kDa N-domain ACE. Methods: To do so, the data from Wistar recipients of kidney from SHR were compared to data from transplanted Wistar submitted to CsA treatment and, to Wistar Sham. Results and Discussion: Despite the unaltered blood pressure observed at early stages, 80/90-kDa ACE was found expressed in the urine of rats 7 and 15 days after transplantation, which was intense when rats became hypertensive 30 days post-surgery. Conclusion: Our data show that this enzyme is associated with the development of hypertension, and this marker appears in the urine before any substantial blood pressure alteration.

Resumo Introdução: A hipertensão é altamente prevalente pós-transplante renal e vários fatores estão associados incluindo o tratamento com imunossupressores e a predisposição genética. Objetivo: Os objetivos foram avaliar os efeitos do transplante do rim de ratos espontaneamente hipertensos (SHR) em ratos Wistar, e a possível transferência da ECA N-domínio de 80/90-kDa para os tecidos dos receptores. Métodos: Para isso, os dados dos animais Wistar receptores dos rins de SHR foram comparados aos dados dos Wistar submetidos ao tratamento com CsA e Wistar Sham. Resultados e Discussão: Apesar da pressão arterial permanecer inalterada nos estágios iniciais pós-transplante renal, a expressão da ECA de 80/90-kDa foi identificada na urina de ratos 7 e 15 dias após o transplante, e de forma mais intensa aos 30 dias após a cirurgia, quando os animais tornaram-se hipertensos. Conclusão: Nossos dados mostram que ECA N-domínio está associada ao desenvolvimento da hipertensão, e que este marcador pode ser identificado na urina pós-transplante renal antes mesmo de qualquer alteração da pressão arterial.

Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in culture.

Angiotensin converting enzyme 2 (ACE2) is a component of the renin-angiotensin system (RAS) which converts Ang II, a potent vasoconstrictor peptide into Ang 1-7, a vasodilator peptide which may act as a negative feedback hormone to the actions of Ang II. The discovery of this enzyme added a new level of complexity to this system. The mesangial cells (MC) have multiple functions in glomerular physiology and pathophysiology and are able to express all components of the RAS. Despite of being localized in these cells, ACE2 has not yet been purified or characterized. In this study ACE2 from mice immortalized MC (IMC) was purified by ion-exchange chromatography. The purified enzyme was identified as a single band around 60-70 kDa on SDS-polyacrylamide gel and by Western blotting using a specific antibody. The optima pH and chloride concentrations were 7.5 and 200 mM, respectively. The N-terminal sequence was homologous with many species ACE2 N-terminal sequences as described in the literature. ACE2 purified from IMC was able to hydrolyze Ang II into Ang 1-7 and the K(m) value for Ang II was determined to be 2.87 ± 0.76 µM. In conclusion, we purified and localized, for the first time, ACE2 in MC, which was able to generate Ang 1-7 from Ang II. Ang 1-7 production associated to Ang II degradation by ACE2 may exert a protective effect in the renal hemodynamic.

Relationship between renal and cardiovascular changes in a murine model of glucose intolerance.

UNLABELLED: Nutrition is an important variable which may affect the risk for renal disease. We previously showed that a high fructose diet in mice produced hypertension and sympathetic activation [8]. The purpose of this study was to determine if a fructose diet altered renal function. A high fructose diet for 12 weeks impaired glucose tolerance, but caused no change in body weight, blood glucose or plasma insulin. Impairment in renal function was documented by the almost two fold increase in urinary protein excretion ( CONTROL: 6.6+/-0.6 vs. Fructose: 15.0+/-0.7 mmol protein/mmol creatinine; p<0.05) which was also accompanied by increases in urinary volume. The diet produced little change in renal histology, kidney weight or kidney weight/body weight ratio. Urinary excretion of angiotensin II/creatinine ( CONTROL: 78.9+/-16.6 vs. Fructose: 80.5+/-14.2 pg/mmol) and renal angiotensin converting enzyme activity ( CONTROL: 9.2+/-1.6 vs. Fructose: 7.6+/-1.0 ACE units) were not different between groups. There was a positive correlation between mean arterial pressure (r=0.7, p=0.01), blood pressure variability (BPV) (r=0.7, p=0.02), low frequency BPV component (r=0.677, p=0.03) and urinary protein excretion. Results show that consumption of a high fructose diet in mice had deleterious effects on renal function, which were correlated with cardiovascular changes.

[Nandrolone administration does not promote hypertrophy of soleus muscle in rats]. / A administração de nandrolona não promove hipertrofia do músculo sóleo em ratos.

Anabolic androgenic steroids (AAS) are compounds formed from testosterone or one of its derivatives, which are largely used by amateur e professional athletes to improve the athletic performance. However, the scientific information about the relation between the use of AAS and muscle hypertrophy is controversial. The aim of this study was to evaluate the effects of testosterone and physical training on muscle hypertrophy. Male Wistar rats received i.m. injections of Deca-Durabolin or vehicle during 6 weeks. Trained rats were submitted to a resistance physical training, by jumping up and down in water carrying an overload. Sedentary and trained animals were anesthetized and sacrificed. Soleus muscle was removed for the quantification of total protein and DNA concentration. In the end of the treatment, body weight of trained animals treated with vehicle or AAS was lower than the body weight of respective sedentary. Total protein concentration and the ratio muscle weight/body weight of all experimental groups were not altered. Trained group treated with AAS presented lower DNA concentration than trained group treated with vehicle. The administration of nandrolone decanoate did not promote hypertrophy on soleus muscle, not even when the use of AAS was associated to resistance physical training.

Nocturnal hypertension in mice consuming a high fructose diet.

OBJECTIVE: To investigate the effect of fructose consumption on the light/dark pattern of blood pressure, heart rate and autonomic neural function in mice. BACKGROUND: Insulin resistant diabetes is associated with hypertension and autonomic dysfunction. There is evidence that the increasing incidence of diabetes may be related to dietary changes, including consumption of high levels of fructose. DESIGN/METHODS: C57/BL mice, instrumented with radiotelemetric arterial catheters, were fed a control or high fructose diet (60%). Cardiovascular parameters measured were light/dark pattern of mean arterial pressure (MAP), heart rate (HR) and variability (time and frequency domain). We also measured plasma insulin, glucose, lipids and angiotensin II (Ang II) as well as glucose tolerance. In situ hybridization was used to measure brainstem expression of tyrosine hydroxylase (TH) and Ang AT1a mRNA. RESULTS: Fructose diet (8 weeks) produced an increase in MAP, variance and low frequency domain (14+/-3 vs. 33+/-4 mm Hg(2), variance and 10+/-2 vs. 26+/-4 mm Hg(2), LF, control vs. fructose, P<0.01). The changes occurred only at night, a period of activity for mice. Glucose tolerance was attenuated in the fructose group. Fructose also increased plasma cholesterol (80+/-1 vs. 126+/-2 mg/dl, control vs. fructose, P<0.05) and plasma Ang II (18+/-5 vs.65+/-12 pg/ml, control vs. fructose, P<0.05). Depressor responses to alpha(1)-adrenergic blockade with prasozin were augmented in fructose-fed mice. Using quantitative in situ hybridization, we found that Ang AT1a receptor and TH mRNA expression were significantly increased in the brainstem locus coeruleus. CONCLUSION: A high fructose diet in mice produced nocturnal hypertension and autonomic imbalance which may be related to activation of sympathetic and angiotensin systems.

A administração de nandrolona não promove hipertrofia do músculo sóleo em ratos / Nandrolone administration does not promote hypertrophy of soleus muscle in rats

Os esteróides anabólicos androgênicos (EAA) são compostos formados a partir da testosterona ou um de seus derivados, sendo amplamente utilizados por desportistas amadores e profissionais com o objetivo de melhorar a performance atlética. Entretanto, a literatura a respeito da relação entre EAA e hipertrofia muscular é controversa. O objetivo deste estudo foi avaliar os efeitos da nandrolona e do treinamento físico sobre a hipertrofia muscular. Ratos Wistar machos receberam injeção i.m. de Deca-Durabolin® ou veículo durante 6 semanas. Os animais dos grupos treinados foram submetidos a treinamento físico resistido, através de sessões de saltos em meio líquido. Os animais sedentários e treinados foram sacrificados após anestesia e o músculo sóleo retirado para quantificação de proteínas totais e DNA. Ao final do tratamento, os animais treinados tratados com veículo ou EAA apresentaram menor peso corporal do que os respectivos grupos sedentários. Não foram observadas diferenças estatísticas na concentração de proteínas totais e na razão peso muscular/peso corporal entre os grupos experimentais. O grupo treinado tratado com EAA apresentou concentração de DNA significativamente menor do que o grupo treinado veículo. A administração de decanoato de nandrolona não promoveu hipertrofia do músculo sóleo, nem mesmo quando associada ao treinamento físico resistido.

New mass spectrometric assay for angiotensin-converting enzyme 2 activity.

A novel assay was developed for evaluation of mouse angiotensin-converting enzyme (ACE) 2 and recombinant human ACE2 (rACE2) activity. Using surface-enhanced laser desorption/ionization time of flight mass spectrometry (MS) with ProteinChip Array technology, ACE1 and ACE2 activity could be measured using natural peptide substrates. Plasma from C57BL/6 mice, kidney from wild-type and ACE2 knockout mice, and rACE2 were used for assay validation. Plasma or tissue extracts were incubated with angiotensin I (Ang I; 1296 m/z) or angiotensin II (Ang II; 1045 m/z). Reaction mixtures were spotted onto the ProteinChips WCX2 and peptides detected using surface-enhanced laser desorption/ionization time of flight MS. MS peaks for the substrates, Ang I and Ang II, and the generated peptides, Ang (1-7) and Ang (1-9), were monitored. The ACE2 inhibitor MLN 4760 (0.01 to 100 micromol/L) significantly inhibited rACE2 activity (IC50=3 nmol/L). Ang II was preferably cleaved by rACE2 (km=5 mumol/L), whereas Ang I was not a good substrate for rACE2. There was no detectable ACE2 activity in plasma. Assay specificity was validated in a model of ACE2 gene deletion. In kidney extract from ACE2-deficient mice, there was no generation of Ang (1-7) from Ang II. However, Ang (1-7) was produced when Ang I was used as a substrate. In conclusion, we developed a specific and sensitive assay for ACE2 activity, which used the natural endogenous peptide substrate Ang II. This approach allows for the rapid screening for ACE2, which has applications in drug testing, high-throughput enzymatic assays, and identification of novel substrates/inhibitors of the renin-angiotensin system.

Relação entre a administração de esteróide anabólico androgênico, treinamento físico aeróbio e supercompensação do glicogênio / Relation between anabolic androgenic steroid administration, aerobic physical training and glycogen supercompensation

A supercompensação do glicogênio é uma das adaptações induzidas pelo treinamento físico. Visando potencializar este fenômeno, muitos atletas utilizam doses suprafisiológicas de esteróides anabólicos androgênicos (EAA). O objetivo deste estudo foi avaliar em ratos os efeitos da nandrolona e do exercício aeróbio sobre o peso corporal, triglicerídeos, glicose e reservas de glicogênio. Ratos Wistar machos foram aleatoriamente divididos em quatro grupos: sedentário + veículo (SV), treinado + veículo (TV), sedentário + EAA (SEAA) e treinado + EAA (TEAA, n = 7-14/grupo). Receberam injeção i.m. de nandrolona ou veículo durante nove semanas e durante o mesmo período os animais treinados foram submetidos a exercício aeróbio. Os dados foram analisados por ANOVA bifatorial e Tukey (p < 0,05). Os grupos SEAA, TV e TEAA apresentaram menor peso corporal do que o grupo SV (SEAA: 339 ± 10 = TV: 342 ± 14 = TEAA: 332 ± 6 < SV: 398 ± 9g). O treinamento físico reduziu significativamente a concentração plasmática de triglicerídeos [(TV: 46 ± 4 = TEAA: 44 ± 3) < (SV: 104 ± 1 = SEAA: 101 ± 6mg/dL)] e de glicogênio hepático [(TV: 3,38 ± 0,57 = TEAA: 2,62 ± 0,34) < (SV: 4,95 ± 0,11 = SEAA: 4,43 ± 0,23mg/100mg)] e aumentou a concentração cardíaca de glicogênio [(TV: 0,38 ± 0,04 = TEAA: 0,42 ± 0,03) > (SV: 0,2 ± 0,02 = SEAA: 0,21 ± 0,02mg/100mg)]. A glicemia e as reservas de glicogênio do sóleo permaneceram inalteradas. O uso de doses suprafisiológicas de nandrolona não potencializou nenhum dos efeitos obtidos em resposta ao treinamento aeróbio.