Regulation of oxidative stress and somatostatin,cholecystokinin, apelin gene expressions by ghrelin in stomach of newborn diabetic rats

The aim of the study was to determine whether ghrelin treatment has a protective effect on gene expression and biochemical changes in the stomach of newborn streptozotocin (STZ) induced diabetic rats. In this study, four groups of Wistar rats were used: control, ghrelin control, diabetic and diabetic + ghrelin. The rats were sacrificed after four weeks of treatment for diabetes. The gene expressions of: somatostatin, cholecystokinin, apelin and the altered active caspase-3, active caspase-8, proliferating cell nuclear antigen, were investigated in the pyloric region of the stomach and antioxidant parameters were measured in all the stomach. Although ghrelin treatment to diabetic rats lowered the stomach lipid peroxidation levels, the stomach glutathione levels were increased. Exogenous ghrelin caused an increased activities of stomach catalase, superoxide dismutase, glutathione reductase and glutathione peroxidase in diabetic rats. Numbers of somatostatin, cholecystokinin and proliferating cell nuclear antigen immunoreactive cells decreased in the diabetic + ghrelin group compared to the diabetic group. Apelin mRNA expressions were remarkably less in the diabetic + ghrelin rats than in diabetic rats. The results may indicate that ghrelin treatment has a protective effect to some extent on the diabetic rats. This protection is possibly accomplished through the antioxidant activity of ghrelin observed in type 2 diabetes. Consequently exogenous ghrelin may be a candidate for therapeutic treatment of diabetes.     

Chard (Beta vulgaris L. var. cicla) extract ameliorates hyperglycemia by increasing GLUT2 through Akt2 and antioxidant defense in the liver of rats

Chard is a plant used as an alternative hypoglycemic agent by diabetic people in Turkey. The aim of this study was to examine the molecular mechanism of hypoglycemic effects of chard extract. Male Sprague-Dawley rats (6–7 months old) were divided into five groups for this investigation: (1) control, (2) hyperglycemic, (3) hyperglycemic + chard, (4) hyperglycemic + insulin, (5) hyperglycemic + chard + insulin. Fourteen days after animals were rendered hyperglycemic by intraperitoneal injection of 60 mg/kg streptozotocin, the chard water extract (2 g/kg/day) or/and insulin (6 U/kg/day) was administered for 45 days. Hypoglycemic effect of chard extract was demonstrated by a significant reduction in the fasting blood glucose and increased glycogen levels in liver of chard extract-treated hyperglycemic rats. Moreover, activity of adenosine deaminase, which is suggested as an important enzyme for modulating the bioactivity of insulin, was decreased by chard treatment. Immunostaining analysis showed increased nuclear translocation of Akt2 and synthesis of GLUT2 in the hepatocytes of chard or/and insulin-treated hyperglycemic rats. The oxidative stress was decreased and antioxidant defense was increased by chard extract or/and insulin treatment to hyperglycemic rats according to the decreased malondialdehyde formation, the activities of catalase, superoxide dismutase, myeloperoxidase and increased glutathione levels. These findings suggest that chard extract might improve glucose response by increasing GLUT2 through Akt2 and antioxidant defense in the liver.     

Protective effects of melatonin against spinal cord injury induced oxidative damage in rat kidney: A morphological and biochemical study

Spinal cord injury (SCI) induced oxidative stress affects multiple organ systems including the kidney. We studied the possible protective effects of melatonin on SCI-induced oxidative damage in renal tissues of rats. Wistar albino rats (n = 24) were exposed to SCI and divided into vehicle- or melatonin-treated SCI groups. Melatonin was administred intraperitoneally at a dose of 10 mg/kg for seven days. Renal tissues were investigated by light and electron microscopy. Furthermore, tissue malondialdehyde (MDA) and glutathione (GSH) levels and myeloperoxidase (MPO) and superoxide dismutase (SOD) activities were also determined. In the vehicle-treated SCI group, the renal histology was disturbed compared to controls, whereas the melatonin-treated SCI group showed significantly reduced degeneration of renal tissue as seen by both light and electron microscopy. MDA levels, MPO and SOD activities were increased and GSH levels were decreased in the vehicle-treated SCI group compared to controls. On the other hand, decreased MDA levels and MPO activities and increased GSH levels were observed in the melatonin-treated SCI group compared to vehicle-treated SCI group. These results showed that experimentally induced SCI caused oxidative stress in the rat kidney, whereas melatonin treatment reduced oxidative stress, suggesting that it may be used as a complementary therapy of renal problems occurring following SCI.     

Extract of Moringa oleifera leaves ameliorates streptozotocin-induced Diabetes mellitus in adult rats

Medicinal plants attract growing interest in the therapeutic management of Diabetes mellitus. Moringa oleifera is a remarkably nutritious vegetable with several antioxidant properties. The present study assessed the possible antioxidant and antidiabetic effects of an aqueous extract of M. oleifera leaves in treating streptozotocin-induced diabetic albino rats. The antidiabetic effects of aqueous extract of M. oleifera leaves were assessed histomorphometrically, ultrastructurally and biochemically. Fasting plasma glucose (FPG) was monitored and morphometric measurements of β-cells of islets of Langerhans (modified Gomori's stain) and collagen fibers (Mallory's trichrome stain) were performed. The antioxidant effects of M. oleifera leaves were determined by measuring the reduced glutathione and lipid peroxidation product, malondialdehyde, in pancreatic tissue. M. oleifera treatment significantly ameliorated the altered FPG (from 380% to 145%), reduced glutathione (from 22% to 73%) and malondialdehyde (from 385% to 186%) compared to control levels. The histopathological damage of islet cells was also markedly reversed. Morphometrically, M. oleifera significantly increased the areas of positive purple modified Gomori stained β-cells (from 60% to 91%) and decreased the area percentage of collagen fibers (from 199% to 120%) compared to control values. Experimental findings clearly indicate the potential benefits of using the aqueous extract of M. oleifera leaves as a potent antidiabetic treatment.     

Role of bone marrow-derived stem cells versus insulin on filiform and fungiform papillae of diabetic albino rats (light,fluorescent and scanning electron microscopic study)

BackgroundDiabetes mellitus (DM) is a chronic metabolic disease characterized by high blood glucose levels. DM affects many body’s organs and caused by insulin production deficiency or by the ineffectiveness of the produced insulin. Administration of exogenous insulin is required for management of type I DM; however, it does not cure the disease. Bone marrow-mesenchymal stem cells (BM-MSCs) have been highlighted to offer a novel cell based approach for treatment of diabetes because of their anti-diabetic effect, direct differentiation into a variety of cell types, or release of paracrine factors.AimTo examine the effect of BM-MSCs versus insulin on true filiform and fungiform papillae of diabetic rats.Materials and methodsFifty six male Wistar albino rats weighing 200–250 g were equally divided into: Control group (Gp I): Rats did not receive any drug. Diabetic group (Gp II): Rats received a single intra-peritoneal injection of streptozotocin (40 mg/kg). BM-MSCs treated diabetic group (Gp III): After DM confirmation; rats received a single intravenous injection of BM-MSCs (million units) through tail vein. Insulin treated diabetic group (Gp IV): After DM confirmation; rats received a daily subcutaneous injection of insulin (5IU/kg). After four weeks, half of the tongue specimens were processed and stained by Hematoxyline & Eosin and Anti-proliferating cell nuclear antigen (Anti-PCNA) then examined by light microscope. Fluorescent microscope was used to detect homing of injected labeled BM-MSCs in rats’ filiform and fungiform papillae. While the other half were examined by scanning electron microscope.ResultsTrue filiform and fungiform papillae of Gp II showed significant histological and morphological alterations. In treated groups, Gp III and Gp IV, both papillae showed marked improvements, being more noticeable in Gp IV. There was a significant increase in the number of Anti-PCNA positive cells and a significant decrease in fasting blood glucose level in Gp III and Gp IV in comparison to Gp II.ConclusionsDM had degenerative effects on true filiform and fungiform papillae. Administration of BM-MSCs reduced the deleterious effects of DM on both papillae. Insulin injection caused more obvious improvements in both papillae of diabetic rats than BM-MSCs.     

Effect of redox status of peripheral blood on immune signature of circulating regulatory and cytotoxic T cells in streptozotocin induced rodent model of type I diabetes

Diabetes mellitus is an autoimmune chronic inflammatory disease manifested by hyperglycemia and associated with imbalance in redox status and inflammatory response. Oxidative stress has been reported to affect functions of T cell repertoire- regulatory T cells (T_regs) and cytotoxic lymphocytes (CTLs). T_regs are involved in prevention against autoreactive T cells and controlling inflammation while CTLs are major mediators of tissue injury. Hence the present study is novel as it contemplates to understand oxidative stress in diabetes vis-à-vis T cells. Comparative analysis was carried out between two groups, i.e., healthy Sprague Dawley (SD) and Streptozotocin (STZ) induced SD rat model of type1 diabetes (T1D). Various hematological, biochemical and oxidative stress parameters were assessed in plasma samples in the study. Peripheral blood mononuclear cells (PBMCs), T_regs and CTLs were evaluated for intracellular oxidative stress using 2′,7′–dichlorofluorescin diacetate (DCFDA), mitochondrial ROS using Mitosox-red, mitochondrial membrane potential using JC-1 in PBMCs. T_reg populations expressing IL-4, IL-6 and IL-10 and CTLs expressing αβ-T cell receptor (αβ -TCR), interferon- γ (IFN-γ), perforin and granzyme were also considered. We found decreased activity of enzymes such as catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and reduced glutathione(GSH) and increased lipid peroxidation (LPO) in plasma indicated altered redox state in diabetic animals. Elevated intracellular reactive oxygen species (ROS) and mitochondrial superoxide was observed in T1D group confirming oxidative stress in cell specific manner. Cell population with hyperpolarized mitochondrial membrane potential was found to be elevated in T1D group. We found a decrease in T_reg population in T1D group in comparison to healthy group. T_reg population expressing IL-4, IL-6 were increased and those expressing IL-10 were found to be reduced in diabetic group. The CTL numbers were dropping whereas αβ-TCR, IFN-γ, perforin and granzyme expressing CTLs were on the rise in diabetic group. Our finding suggested an increased oxidative stress in T_regs and CTLs which might be responsible for progressive inflammatory environment built up due to persistent hyperglycemia. This was fortified by the statistical analyses where strong correlation between LPO and CTLs expressing TCR, IFN-γ, perforin and granzyme was noted. Lipid peroxidation was also found to be correlated to intracellular ROS in T_regs and CTLs along with other important revelations. The present study gives important insights into the significance of oxidative stress on immune system and its mediators in diabetes.