In utero protein restriction causes growth delay and alters sperm parameters in adult male rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diabetes evolution in rats after neonatal treatment with alloxan

Physical exercises have been recommended in the prevention of non-insulin dependent diabetes mellitus (NIDDM), but the mechanisms involved in this intervention are not yet fully understood. Experimental models offer the opportunity for the study of this matter. The present study was designed to analyze the diabetes evolution in rats submitted to neonatal treatment with alloxan with the objective of verifying the suitability of the model to future studies with exercises. For this, newly born rats (6 days old) received intraperitoneal alloxan (A = 200 mg/kg of body weight). Rats injected with vehicle (citrate buffer) were used as controls (C). The fasting blood glucose level (mg/dL) was higher in the alloxan group at the day 28 (C=47.25 +/- 5.08; A=54.51 +/- 7.03) but not at the 60 day of age (C=69.18 +/- 8.31; A=66.81 +/- 6.08). The alloxan group presented higher blood glucose level during glucose tolerance test (GTT) (mg/dL. 120 min) in relation to the control group both at day 28 (C=16908.9 +/- 1078.8; A=21737,7 +/- 1106.4) and at day 60 (C=11463.45 +/- 655.30; A=15282.21 +/- 1221.84). Insulinaemia during GTT (ng/mL.120 min) was lower at day 28 (C=158.67 +/- 33.34; A=123.90 +/- 19.80), but presented no difference at day 60 (C=118.83 +/- 26.02; A=97.8 +/- 10.88). At day 60, the glycogen concentration in the soleus muscle (mg/100mg) was lower in the alloxan group (0.3 +/- 0.13) in relation to the control group (0.5 +/- 0.07). No difference was observed between groups in relation to (mu mol/g.h): Glucose Uptake (C = 5.8 +/- 0.63; A = 5.2 +/- 0.73); Glucose Oxidation (C= 4.3 +/- 1.13; A= 3.9 +/- 0.44); Glycogen Synthesis (C= 0.8 +/- 0.18; A= 0.7 +/- 0.18) and Lactate Production (C= 3.8 +/- 0.8; A= 3.8 0.7) by the isolated soleus muscle. The glucose-stimulated insulin secretion (16.7mM) by the isolated islets (ng/5 islets. h) of the alloxan group was lower (14.3 +/- 4.7) than the control group (32.0 +/- 7.9). Thus, we may conclude that this neonatal diabetes induction model gathers interesting characteristics and may be useful for further studies on the role of the exercise in the diabetes mellitus appearance.

Different exercise protocols improve metabolic syndrome markers, tissue triglycerides content and antioxidant status in rats

Background: An increase in the prevalence of obesity entails great expenditure for governments. Physical exercise is a powerful tool in the combat against obesity and obesity-associated diseases. This study sought to determine the effect of three different exercise protocols on metabolic syndrome and lipid peroxidation markers and the activity of antioxidant enzymes in adult Wistar rats (120 days old).Methods: Animals were randomly divided into four groups: the control (C) group was kept sedentary throughout the study; the aerobic group (A) swam1 h per day, 5 days per week, at 80% lactate threshold intensity; the strength group (S) performed strength training with four series of 10 jumps, 5 days per week; and the Concurrent group (AS) was trained using the aerobic protocol three days per week and the strength protocol two days per week.Results: Groups A and S exhibited a reduction in body weight compared to group C. All exercised animals showed a reduction in triglyceride concentrations in fatty tissues and the liver. Exercised animals also exhibited a reduction in lipid peroxidation markers (TBARS) and an increase in serum superoxide dismutase activity. Animals in group A had increased levels of liver catalase and superoxide dismutase activities.Conclusions: We concluded that all physical activity protocols improved the antioxidant systems of the animals and decreased the storage of triglycerides in the investigated tissues.

Effects of experimental osteoporosis and low calcium intake on postextraction sockets of rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effects of the administration of a catalase inhibitor into the fourth cerebral ventricle on cardiovascular responses in spontaneously hypertensive rats exposed to sidestream cigarette smoke

OBJECTIVE: ,,,,,Previous studies have demonstrated a relationship between brain oxidative stress and cardiovascular regulation. We evaluated the effects of central catalase inhibition on cardiovascular responses in spontaneously hypertensive rats exposed to sidestream cigarette smoke. ,,,, ,,,, ,,,,,METHODS: ,,,,,Male Wistar Kyoto (WKY) rats and spontaneously hypertensive rats (SH) (16 weeks old) were implanted with a stainless steel guide cannula leading into the fourth cerebral ventricle (4th V). The femoral artery and vein were cannulated for arterial pressure and heart rate measurement and drug infusion, respectively. The rats were exposed to sidestream cigarette smoke for 180 minutes/day, 5 days/week for 3 weeks (CO: 100-300 ppm). The baroreflex was tested using a pressor dose of phenylephrine (8 μg/kg, bolus) and a depressor dose of sodium nitroprusside (50 μg/kg, bolus). Cardiovascular responses were evaluated before and 5, 15, 30 and 60 minutes after injection of a catalase inhibitor (3-amino-1,2,4-triazole, 0.001 g/100 μL) into the 4th V. ,,,, ,,,, ,,,,,RESULTS: ,,,,,Vehicle administration into the 4th V did not affect the cardiovascular response, whereas administration of the central catalase inhibitor increased the basal HR and attenuated the bradycardic peak (p<0.05) to a greater extent in WKY rats exposed to sidestream cigarette smoke than in WKY rats exposed to fresh air. However, in spontaneously hypertensive rats, the effect of the catalase inhibitor treatment was stronger in the fresh air condition (p<0.05). ,,,, ,,,, ,,,,,CONCLUSION: ,,,,,Administration of a catalase inhibitor into the 4th V combined with exposure to sidestream cigarette smoke has a stronger effect in WKY rats than in SH rats.

Long term evaluation of morphometric and ultrastructural changes of testes of alloxan-induced diabetic rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The role of exercise on long-term effects of alloxan administered in neonatal rats

The present study was designed to analyse the effects of aerobic exercise on the metabolic effects of alloxan. Male Wistar newborn rats (2 days old) received alloxan (200 mg (kg body weight)(-1)) intraperitoneally (A rats). Vehicle-injected rats were used as controls (C rats). At 28 days old, some of the A rats were subjected to swimming for 1 h day(-1), 5 day week(-1) (AT rats). At 28, 60 and 90 days old the animals were subjected to glucose (GTTo) and insulin (ITTsc) tolerance tests. All the animals were then killed by decapitation for blood and tissue evaluations. on the 60th day, there was a reduction in blood glucose level during the GTTo (mmol l(-1) (90 min)(-1)) in the AT rats (7640.7+/-694.0) with respect to C (7057.5+/-776.9) and A (8555.6+/-1096.7) rats. However on the 90th day, AT rats showed higher glucose levels (8004.6+/-267.9) when compared to the other groups (C, 7305.5+/-871.2; A, 7088.8+/-536.9). The serum free fatty acid (FFA) concentration (muEq l(-1)) was higher in the alloxan-treated animals (A, 231.1+/-58.5; AT, 169.8+/-20.1) than in controls (C, 101.4+/-22.4). In conclusion, although the high blood glucose level is transitory in the A animals, some blood and tissue alterations remain and can be harmful to the maintenance of homeostasis. Physical exercise counteracted only partially these alterations. Furthermore, training worsened glucose tolerance at the 90th day, suggesting that exercise intensity should be adjusted to the diabetic condition.

Effect of long-term treatment with insulin and/or acarbose on glomerular basement membrane thickening in alloxan-diabetic rats

Acarbose is a competitive inhibitor of the intestinal alpha-glycosidases, that can delay absorption of intestinal carbohydrates causing their malabsorption. In the present paper we studied the effects of insulin, acarbose and their association on glomerular basement membrane thickening in alloxan-diabetic rats. Twenty-five male and female Wistar rats, approximately 3 months old at the beginning of the experiment, were assigned randomly to each of five experimental groups: normal control rats, alloxan-diabetic control rats, alloxan-diabetic rats treated with acarbose, alloxan-diabetic rats treated with insulin, and alloxan-diabetic rats treated with insulin plus acarbose. Alloxan was administered in a single iv dose of 42 mg/kg body weight. Insulin was given subcutaneously at doses of 18 to 30 IU/kg corrected daily on the basis of glycosuria and ketonuria. Acarbose was given mixed with rat chow in a dose of 50 mg/100 g chow. Body weight, water and food intake and diuresis, as well as blood and urine glucose were determined after 1, 3, 6, 9, and 12 months of treatment. Glomerular basement membrane (GBM) thickening was determined by electron microscopy at the same times. Clear clinical and laboratory signs of severe diabetes, with blood glucose levels above 200 mg/dl and urine glucose above 3000 mg/dl, were observed in all alloxan-diabetic control rats, in all periods of follow-up, whereas administration of insulin or acarbose reduced the blood glucose levels of treated groups. The most satisfactory control of blood and urine glucose was observed in animals treated with both insulin and acarbose. However, diarrhea was observed in diabetic rats treated with acarbose associated or not with insulin. GBM thickening was correlated with age in all groups. Beginning at six months after diabetes induction, the GBM of untreated diabetic rats was significantly thicker (mean +/- SEM, 4.446 +/- 0.45 mm) than that of normal rats (2.977 +/- 0.63 mm). Both insulin and acarbose prevented GBM thickening and their combination induced thickening similar to the age-dependent thickening observed for normal rats of the same age. We conclude that acarbose when combined with insulin may be a good option in the control of diabetes and its renal complications.

EFFECTS OF PROTEIN-DEFICIENCY AND NATURAL INTESTINAL INFECTION WITH GIARDIA-LAMBLIA ON JEJUNAL INTRAEPITHELIAL LYMPHOCYTES IN RATS OF DIFFERENT AGES

1. The interaction between experimental protein deprivation and natural intestinal infection by Giardia lamblia was studied in terms of its effects on the intraepithelial lymphocyte (IEL) population and morphology of the jejunal mucosa of rats of different ages.2. Young, adult and old male Wistar rats received a protein-deficient diet (2% casein) or a control diet (20% casein) for 42 days. Mucosal height and the number of lymphocytes located among 500 consecutive epithelial cells (EC) along the villi or crossing the basement membrane were determined in PAS-stained jejunal fragments.3. The number of IEL increased progressively with animal age, from 14 to 25 per 100 epithelial cells, with significant differences between age ranges. However, the number of IEL did not differ between control and protein-deficient rats in any of the age groups. The proportion of lymphocytes crossing the basement membrane was approximately two-fold greater in young (2.8/100 EC) and adult (5.8/100 EC) protein-deficient animals than in their respective controls (1.6 and 2.8/100 EC). The intensity of parasite colonization was moderate, from 3 to 5/100 EC and did not differ between groups. The pattern of morphologic changes of jejunal mucosa in protozoal infection did not differ between control and protein-deficient animals in any of the three age groups.4. We conclude that intestinal infection with Giardia lamblia probably stimulated the local immune response, masking the reduction of the IEL population induced by protein deficiency. The increase in lymphocyte numbers with age may be related to prolonged antigenic stimulation promoted by infection.

Anthropometrical parameters and markers of obesity in rats

The present study was undertaken to determine anthropometrical parameters in male adult Wistar rats. We tested the hypothesis that the anthropometrical index may identify obesity and may predict its adverse effects on lipid profile and oxidative stress in rats. Two experimental protocols were performed. in the first experiment, 50 male Wistar rats, 21 days old and fed a control chow were studied up to 150 days of age. in the second experiment, male Wistar rats, 60 days old, were divided into three groups (n = 8): control (C) given free access to a control chow; (S) receiving the control chow and drinking 30% sucrose ad libitum and (HQ fed a high-carbohydrate diet ad libitum. The first experiment showed that food consumption, energy intake and body weight increased with increasing age, while specific rate of body mass gain was significantly decreased. There were no significant differences in body length and thoracic circumference of rats from 60 days of age. The abdominal circumference (AC) and body mass index (BMI) significantly increased with enhancing age in rats up to 90 days of age and remained constant thereafter. In the second experiment, after 30 days of dietary treatment, the final body weight, body mass gain, carcass fat and BMI were higher in S and HC rats than in C. There were no significant alterations in body length and carcass protein among the groups. Triacylglycerol (TG), total cholesterol (CT), low-density lipoprotein cholesterol (LDL-C) and lipid hydroperoxide (LH) were higher in S and HC rats than in C. High-density lipoprotein cholesterol (HDL-C) decreased in HC rats and total antioxidant substances (TAS) decreased in S and HC rats. There were positive correlations between BMI with carcass fat, BMI with LH and BMI and serum TG concentration. In conclusion, the BMI for male adult Wistar rats ranged between 0.45 and 0.68 g/cm(2). Obesity may be easily estimated from the BMI in rats. Alterations in BMI were associated with dyslipidemic profile and oxidative stress in serum of rats and BMI may predict these adverse consequences of the obesity in rats.

EFFECT OF CAFFEINE ON THE METABOLISM OF RATS EXERCISING BY SWIMMING

Several studies have demonstrated that caffeine improves endurance exercise performance but the mechanisms are not fully understood. Possibilities include increased free fatty acid (FFA) oxidation with consequent sparing of muscle glycogen as well as enhancement of neuromuscular function during exercise. The present study was designed to investigate the effects of caffeine on liver and muscle glycogen of 3-month old, male Wistar rats (250-300 g) exercising by swimming. Caffeine (5 mg/kg) dissolved in saline (CAF) or 0.9% sodium chloride (SAL) was administered by oral intubation (1 mu l/g) to fed rats 60 min before exercise. The rats (N = and-IO per group) swam bearing a load corresponding to 5% body weight for 30 or 60 min. FFA levels were significantly elevated to 0.475 +/- 0.10 mEq/l in CAF compared to 0.369 +/- 0.06 mEq/l in SAL rats at the beginning of exercise. During exercise, a significant difference in FFA levels between CAF and SAL rats was observed at 30 min (0.325 +/- 0.06 vs 0.274 +/- 0.05 mEq/l) but not at 60 min (0.424 +/- 0.13 vs 0.385 +/- 0.10 mEq/l). Blood glucose showed an increase due to caffeine only at the end of exercise (CAF = 142.1 +/- 27.4 and SAL = 120.2 +/- 12.9 mg/100 ml). No significant difference in liver or muscle glycogen was observed in CAF as compared to SAL rats, at rest or during exercise. Caffeine increased blood lactate only at the beginning of exercise (CAF = 2.13 +/- 0.2 and SAL = 1.78 +/- 0.2 mmol/l). These data indicate that caffeine (5 mg/kg) has no glycogen-sparing effect on rats exercising by swimming even though the FFA levels of CAF rats were significantly higher at the beginning of exercise.

Differential vascular adaptive response in spontaneously hypertensive and Wistar rats: Role of nitric oxide, and prehypertensive and hypertensive states

Stress-induced vascular adaptive response in SHR was investigated, focusing on the endothelium. Noradrenaline responses were studied in intact and denuded aortas from 6-week-old (prehypertensive) and 14-week-old (hypertensive) SHR and age-matched Wistar rats submitted or not to acute stress (20-min swimming and I-h immobilization 25 min apart), preceded or not by chronic stress (2 sessions 2 days apart of 1-h day immobilization for 5-consecutive days). Stress did not alter the reactivity of denuded aorta. Moreover, no alteration in the EC50 values was observed after stress exposure. In intact aortas, acute stress-induced hyporeactivity to noradrenaline similar between strains at both age. Chronic stress potentiated this adaptive response in 6- and 14-week-old Wistar but not in 6-week-old SHR, and did not alter the reactivity of 14-week-old SHR. Maximum response (g) in intact aortas [6-week-old: Wistar 3.25 +/- 0.12, Wistar/acute 1.95 +/- 0.12*, Wistar/chronic 1.36 +/- 0.21*(+), SHR 1.75 +/- 0.11, SHR/acute 0.88 +/- 0.08*, SHR/chronic 0.85 +/- 0.05*; 14-week-old: Wistar 3.83 +/- 0.13, Wistar/acute 2.72 +/- 0.13*, Wistar/chronic 1.91 +/- 0.19*', SHR 4.03 +/- 0.17, SHR/acute 2.26 +/- 0.12*, SHR/chronic 4.10 +/- 0.23; inside the same strain: *P < 0.05 relate to non-stressed rat, (+)P < 0.05 related to acute stressed rat; n = 6-18]. Independent of age and strain, L-NAME and endothelium removal abolished the stress-induced aorta hyporeactivity. Conclusion: the vascular adaptive response to stress is impaired in SHR, independently of the hypertensive state. Moreover, this vascular adaptive response is characterized by endothelial nitric oxide-system hyperactivity in both strains. (c) 2006 Elsevier B.V. All rights reserved.

Free activity in the cage associated with body weight and restoration of bone structural and mechanical properties in growing rats after hindlimb unloading

We investigated the importance of daily free activity in the cage and body weight gain during the recovering of bone structural and mechanical properties in growing rats after hindlimb unloading. Eight-week-old male Wistar rats were randomly divided into control (CG, n=24) and suspended (SG, n=24) groups. Animals from SG underwent a four-week hindlimb unloading period by tail-suspension. Animals from CG and those from SG after release were kept in collective cages and sacrificed at the age of 12, 16 and 20 weeks. Both femurs were removed and its area, bone mineral density (BMD), resistance to failure and stiffness were determined. Four-week hindlimb unloading decreased (p < 0.05) body weight (CG, 373.00 +/- 9.47 vs. SG, 295.86 +/- 9.19 g), BMD (CG, 0.19 +/- 0.01 vs. SG, 0.15 +/- 0.01 g/cm(2)), bone resistance to failure (CG, 147.75 +/- 5.05 vs. SG, 96.40 +/- 5.95 N) and stiffness (CG, 0.38 +/- 0.01 vs. SG, 0.23 +/- 0.02 N/m). Eight weeks of free activity in cage recovered (p > 0.05) the body weight (CG, 472.75 +/- 14.11 vs. SG, 444.75 +/- 18.91 g), BMD (CG, 0.24 +/- 0.01 vs. SG, 0.22 +/- 0.01 g/cm(2)), bone resistance to failure (CG, 195.73 +/- 10.06 vs. SG, 178.45 +/- 8.48 N) and stiffness (CG, 0.56 +/- 0.02 vs. SG, 0.47 +/- 0.03 N/m) of SG animals. Body weight correlated strongly with bone structural and mechanical properties (p < 0.0001). In conclusion, free activity in the cage associated with body weight gain restored bone structural and mechanical properties in growing rats after hindlimb unloading.

Cardiac benefits of exercise training in aging spontaneously hypertensive rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Creatine does not promote hypertrophy in skeletal muscle in supplemented compared with nonsupplemented rats subjected to a similar workload

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)