Alloxan diabetes alters the tensile strength, morphological and morphometric parameters of abdominal wall healing in rats

PURPOSE:To investigate the effects of alloxan diabetes on the abdominal wall healing of rats undergoing laparotomy.METHODS:Ninety-six male Wistar rats weighing between 200 and 300 grams, divided into two groups: non-diabetic group (G1) and another with untreated diabetes (G2). Three months after diabetes induction, the animals underwent a 5cm-long- laparotomy and 5.0 nylon monofilament suture. After the surgery, 12 animals from each group were euthanized on days 4, 14, 21 and 30 corresponding to the moments M1, M2, M3 and M4. In each moment a fragment of the abdominal wall containing the scar was removed for tensile strength measurement, histological and morphometric study. Clinical and biochemical parameters were also analyzed.RESULTS:G2 animals showed parameters compatible with severe diabetes and decreased plasma levels of insulin. The tensile strength in G2 was significantly smaller in M2 and M4, with a tendency to fall in the other two. Through light microscope, diabetic animals showed more difficulty to increase collagen density and contraction. G2 animals showed high cellularity of fibroblasts in later healing moments, with collagen thinning in M2 and M4.CONCLUSION:The abdominal wound healing in untreated diabetic animals was altered and led to a higher incidence of dehiscence and infections.

Effects of protein malnutrition on glucose tolerance in rats with alloxan-induced diabetes

Protein-calorie malnutrition produces glucose intolerance and reduced insulin release in response to glucose. Rats adapted to low- or high-protein diets show an increased resistance to the diabetogenic action of a single dose of streptozotocin or alloxan. To determine the effects of dietary protein level on pancreatic function, we measured serum glucose levels under basal conditions and during the oral glucose tolerance test (GTT) performed before and after a single dose of alloxan administered to rats fed a 25% or a 6% protein diet for a period of 8 weeks. The incidence of mild hyperglycemia (serum glucose > 250 mg/dl) was greater among the rats fed the 25% protein diet (81%) than among those fed the 6% protein diet (42%). During the GTT performed before alloxan administration the serum glucose levels of the rats fed the 6% protein diet were not found to be significantly different from those of rats fed the 25% protein diet. During the GTT performed after alloxan injection all rats showed intolerance to the substrate (serum glucose > 160 mg/dl 120 min after glucose administration) regardless of whether basal serum glucose was normal or high. In summary, alloxan was less effective in producing basal hyperglycemia in the rats fed the 6% protein diet than in those fed the 25% protein diet but caused glucose intolerance during the oral GTT in both groups. Thus, it seems that feeding a 6% protein diet to rats offers only partial protection against the toxic effects of alloxan.

Diabetes mellitus triggers oxidative stress in the liver of alloxan-treated rats: A mechanism for diabetic chronic liver disease

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Anastomotic healing in ileum and colon of alloxan-induced diabetic rats

OBJETIVO: Investigar se o diabetes mellitus pode alterar a força de ruptura (FR) e o conteúdo de colágeno em anastomoses realizadas no íleo e cólon de ratos. MÉTODOS: 300 ratos Wistar foram distribuídos por sorteio em 5 grupos experimentais com 60 animais cada: controle normal manipulado cirurgicamente (G1); normais controles submetidos a anastomoses no íleo (G2) e cólon (G3); ratos diabéticos submetidos a anastomoses no íleo (G4) e cólon (G5). Cada grupo foi dividido em 6 subgrupos com 10 ratos cada para sacrifícios com 0, 4, 7, 14, 21 e 30 dias após as operações. Os procedimentos cirúrgicos foram realizados 3 meses após a indução do diabetes com aloxana. A FR foi medida em todas anastomoses intestinais. Fragmentos de anastomoses do íleo e cólon foram retirados para dosagens de hidroxiprolina (HP) e proteína tecidual total (PT). RESULTADOS: A FR teve significante redução (P<0,05) nos grupos diabéticos G4 e G5, até 7 e 14 dias após a operação, respectivamente, quando comparada à observada nos grupos controles G2 e G3. Não foram observadas diferenças significantes nas dosagens de HP e PT em ratos diabéticos e controles, tanto operados no íleo como no cólon, em todos os momentos de avaliação. CONCLUSÃO: O diabetes conduz a alterações da força de ruptura de anastomoses intestinais durante a fase inicial da reparação da ferida cirúrgica, porém, este fato parece não estar relacionado à capacidade de sintetizar colágeno.

Temporal relationship between successful pancreas transplantation and control of ocular complications in alloxan-induced diabetic rats

Purpose. The impact of pancreas transplantation (PT) on the progression of eye disease is still controversial. This study evaluated the course of retinopathy in transplanted rats in two different diabetic stages.Methods. Sixty inbred male Lewis rats were assigned to four experimental groups: NC-15 nondiabetic control rats; DC-15 untreated diabetic control rats; PT1-15 diabetic rats that received syngeneic pancreas transplants 2 weeks after alloxan diabetes induction; PT2-15 diabetic rats that received pancreas transplants 12 weeks after diabetes onset. Clinical and laboratory parameters and tens opacity were examined in all rats prior to treatment and at 1-, 6-, and 12-months follow-up. Nucleated eyes from five rats in each group processed for ultrastructural study of the retinal at 6 and 12 months after PT or at follow-up.Results. Cataracts were observed in 20%, 60%, and 100% of DC rats at 1-, 6-, and 12-months follow-up, respectively. Early PT (2 weeks) significantly reduced the prevalence of this complication but not late (12 weeks) PT. PT1 rats also showed improved ultrastructure of the superficial and deep capillary plexuses of the retina, and of Muller cells, compared with DC and PT2. In the last group, retinopathy continued to evolve despite successful PT.Conclusion. Our results suggested that prevention of diabetic ocular lesions by PT was closely dependent on earlier performance of the procedure.

Malignant lesions in the ventral prostate of alloxan-induced diabetic rats

The aim of this study was to evaluate the changes caused by chronic diabetes in the rat ventral prostate and to establish a correlation between diabetes and the development of prostatic lesions. Male rats received alloxan (42 mg/kg b.w.) to induce diabetes. Ninety days after diabetes diagnosis, animals were sacrificed and the ventral prostate was removed and prepared for general and immunohistochemical analyses. The total area showing different types of lesions was estimated. Diabetes led to a decrease in the body and prostatic weights, as well as in testosterone levels. The prostate morphology and stereology showed high variation in the diabetic group. Some animals had light changes; the great majority had an intense epithelial atrophy; and other rats showed premalignant and malignant lesions in the prostate. Such epithelial atrophy was, in some samples, combined with chronic inflammation, similar to proliferative inflammatory atrophy (PIA). The diabetic group also presented high incidence of prostatitis, adenocarcinoma and prostatic intra-epithelial neoplasia (PIN). Samples with adenocarcinoma had poorly differentiated acini with high levels of cellular proliferation and nuclear atypia. These lesions exhibited an invasive feature showing Bcl-2-positive cells and interruptions in the basement membrane. An association of PIA, PIN and adenocarcinoma was detected in one sample. Reduced androgen levels have a synergic effect to insulin dysfunction promoting negative effects in the rat prostate. Diabetic individuals had a high incidence of prostatitis, and this inflammation could stimulate the incidence of other forms of prostatic pathology.

Efeitos em curto prazo da aplicação de aloxana para indução de diabetes em ratos wistar

The present study aimed to verify action of alloxan in metabolic and immune parameters after 24 and 192 hours of the injection in Wistar rats. Thus, eight rats were fasted and received monohidrated alloxan Sigma (32 mg/kg body weight) via endovenous. Glycemia and trglyceridemia analyzes were performed before and 192 hours after alloxan application. After 24 hours, alloxan application increased water intake and decreased body mass, food intake and leucocytes counting. 192 hours after alloxan application, there was a recuperation in food intake and leucocytes counting. on the other hand, in this period there was an increase of glycemia and water intake and reduction of body mass. These results indicate that some of diabetic signs caused by alloxan occur in short-term after drug administration.

Effects of acute cold exposure on rectal temperature, blood glucose and plasma free fatty acids in alloxan-diabetic rats

These experiments were carried out to study the effects of acute cold exposure (0-2°C/4 hr) on rectal temperature, blood glucose and plasma free fatty acids (FFA) in alloxan-diabetic rats. Male Wistar rats weighing 170-190 g were used and diabetes was induced by i.v. alloxan injection (40 mg/kg body wt). Cold exposure produced severe hypothermia in diabetic rats. After 4 hr of cold, blood glucose of diabetic rats was reduced from 296±16 to 86t±12 mg/dl (P<0.01), and FFA increased slightly, but was not statistically different (P>0.05) from the initial value. As expected, interscapular brown adipose tissue (IBAT) and retroperitoneal and epididymal white adipose tissues were significantly lower in diabetic than in control rats. Cold exposure reduced total IBAT lipids in control but not in diabetic animals. The results of this experiment suggest that diabetic rats were unable to maintain body temperature in the cold, probably because of a failure to generate an adequate amount of heat by nonshivering thermogenesis in brown adipose tissue.

O RATO COMO MODELO EXPERIMENTAL DO DIABETES - ESTUDO CLINICO-LABORATORIAL

In this study, 90 Wistar rats were used. They were equally divided into three experimental groups - control group (CG), diabetic group (DG) and treated diabetic group (TG). The analyzed parameters were clinical (behavior, activity, general aspect, weight, water ingestion and diuresis) and biochemical (fasting glycemia and urinary glycosis). The diabetes was induced by alloxan and, then, treated with insulin associated to oral hypoglycemic (acarbosis). Observations were made at 5 experimental moments, as it follows: 1, 3, 6, 9, and 12 months after the diabetes induction. The results were submitted to variance analysis, with 5% of significance level. The DG presented lower weight and higher diuresis level than the CG and TG. The water ingestion of the CG was similar to TG. The glycemia levels were higher in DG than in CG, at every experimental moment. The TG, however, presented glycemia similar to the CG, except for the dosages at 3, and 9 months. They urinary glycosis of the DG and TG were similar between themselves, but higher than the one of the CG.

Protein Deficiency Attenuates the Effects of Alloxan on Insulin Secretion and Glucose Homeostasis in Rats

We have investigated the effect of alloxan on insulin secretion and glucose homeostasis in rats maintained on a 17% protein (normal protein, NP) or 6% protein (low protein, LP) diet from weaning (21 days old) to adulthood (90 days old). The incidence of alloxan diabetes was higher in the NP (3.5 times) than in the LP group. During an oral glucose tolerance test, the area under serum glucose curve was lower in LP (57%) than in NP rats while there were no differences between the two groups in the area under serum insulin curve. The serum glucose disappearance rate (Kitt) after exogenous insulin administration was higher in LP (50%) than in NP rats. In pancreatic islets isolated from rats not injected with alloxan, acute exposure to alloxan (0.05 mmol/L) reduced the glucose- or arginine-stimulated insulin secretion of NP islets by 78% and 56%, respectively, whereas for islets from LP rats, the reduction was 47% and 17% in the presence of glucose and arginine, respectively. Alloxan treatment reduced the glucose oxidation in islets from LP rats to a lesser extent than in NP islets (23% vs. 56%). In conclusion, alloxan was less effective in producing hyperglycemia in rats fed a low protein diet than in normal diet rats. This effect is attributable to an increased peripheral sensivity to insulin in addition to a better preservation of glucose oxidation and insulin secretion in islets from rats fed a low protein diet.

Exercise training in the aerobic/anaerobic metabolic transition prevents glucose intolerance in alloxan-treated rats

Background: Ninety percent of cases of diabetes are of the slowly evolving non-insulin-dependent type, or Type 2 diabetes. Lack of exercise is regarded as one of the main causes of this disorder. In this study we analyzed the effects of physical exercise on glucose homeostasis in adult rats with type 2 diabetes induced by a neonatal injection of alloxan. Methods: Female Wistar rats aged 6 days were injected with either 250 mg/ kg of body weight of alloxan or citrate buffer 0.01 M (controls). After weaning, half of the animals in each group were subjected to physical training adjusted to meet the aerobic-anaerobic metabolic transition by swimming 1 h/day for 5 days a week with weight overloads. The necessary overload used was set and periodically readjusted for each rat through effort tests based on the maximal lactate steady state procedure. When aged 28, 60, 90, and 120 days, the rats underwent glucose tolerance tests (GTT) and their peripheral insulin sensitivity was evaluated using the HOMA index. Results: The area under the serum glucose curve obtained through GTT was always higher in alloxan-treated animals than in controls. A decrease in this area was observed in trained alloxan-treated rats at 90 and 120 days old compared with non-trained animals. At 90 days old the trained controls showed lower HOMA indices than the non-trained controls. Conclusion: Neonatal administration of alloxan induced a persistent glucose intolerance in all injected rats, which was successfully counteracted by physical training in the aerobic/anaerobic metabolic transition. © 2008 Mota et al; licensee BioMed Central Ltd.

Zinc sulphate administered by transdermal iontophoresis improves breaking strength of surgical wounds in skin of alloxan-induced diabetic rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

MULLER CELLS AND DIABETIC-RETINOPATHY

Muller cells provide nutrition for neural cells. We studied the structure and ultrastructure of Muller cells in the retina of thirty 3-month old Wistar rats; divided equally into 3 groups: normal rats, alloxan diabetic rats and treated alloxan diabetic rats. 1 and 12 months after induction of diabetes. We observed that the Muller cell nuclei under light microscope examination had hexagonal shape and higher density than the other nuclei. Differences between groups could be observed only by electron microscopy. In the diabetic rats, Muller cells presented dispersion of nuclear chromatin and electrondense nuclear granulations, with the presence of increased glycogen, dense bodies and lysosomes in the cytoplasm. The alterations were more frequent in the perivascular region and at 12 months. The treated diabetic rats exhibited some alterations we observed in diabetic rats. but these alterations were less intense. We conclude that, despite the treatment, the diabetic retinopathy continues to evolve.

Acute and short-term insulin-induced molecular adaptations of GLUT2 gene expression in the renal cortex of diabetic rats

Increased GLUT2 gene expression in the renal proximal tubule of diabetic rats is an adaptive condition, which may be important in the diabetic nephropathy development. We investigated the effects of insulin treatment upon the renal GLUT2 overexpression of diabetic rats. Acute treatment, surprisingly, induced a rapid further increase in GLUT2 mRNA content. Twelve hours after insulin injection, GLUT2 mRNA was twice the value of saline-injected rats (P < 0.001), when GLUT2 protein remained unchanged. In response to short-term treatment, both GLUT2 mRNA and protein were increased in 1-day treated rats (P < 0.05 versus saline-injected), decreasing after that, and reaching, within 6 days, values close to those of non-diabetic rats. Concluding, insulin treatment induced: initially, an additional upregulation of GLUT2 gene expression, involving posttranscriptional modulation; thereafter, downregulation of GLUT2 expression, which returns to non-diabetic levels. The former may be related to increased insulin concentration, the latter may be due to glycemic control. © 2005 Elsevier B.V. All rights reserved.

The number of podocyte and slit diaphragm is decreased in experimental diabetic nephropathy

Purpose: To determine the number of podocyte, slit diaphragms, slit diaphragm extensions and GBM thickness in diabetic nephropathy. Methods: Sixty Rattus Wistarof both sexes weighing 200-300g were divided in two experimental groups: normal group 10 animals, and alloxan diabetic rats - 50 animals. Alloxan was administered in a single IV dose of 42mg/kg body weight. Body weight, water and food intake, diuresis, and blood and urine glucose were determined in both groups before alloxan injection and two weeks, six and twelve months after alloxan injection. Proteinuria was measured at 12 months in both groups. After 12 months animals were sacrificed, and the right kidney processed for electron microscopy. Results: Clear clinical and laboratory signs of severe diabetes were seen, in all alloxan-diabetic rats at all follow-up times. Glomerular basement membrane (GBM) thickening, podocyte number, and slit diaphragm number and extension were determined. GBM of all diabetic rats was significantly thicker (median=0.29μm; semi-interquartile range=0.065μm) than in the normal rats (0.23μm; 0.035μm). Diabetic rat podocyte number (8; 1), slit diaphragm number (4; 1), and slit diaphragm extension (0.021μm; 0.00435μm) were significantly lower than in normal rats (11; 1) and (7; 1.5), and (0.031μm; 0.0058μm). Diabetic rat proteinuria (0.060mg/24h; 0.037mg/24h) was higher than in normal rats (0.00185mg/24h; 0.00055mg/24h). Conclusion: Experimental diabetes is associated with significant (p<0.05) changes in podocyte foot process, slit number, slit diaphragm extension, and GBM thickness.