STZ-induced Damage Research Articles

Lentinan protects pancreatic β cells from STZ-induced damage.

Pancreatic β‐cell death or dysfunction mediated by oxidative stress underlies the development and progression of diabetes mellitus (DM). In this study, we evaluated the effect of lentinan (LNT), an active ingredient purified from the bodies of Lentinus edodes, on pancreatic β‐cell apoptosis and dysfunction caused by streptozotocin (STZ) and the possible mechanisms implicated. The rat insulinoma cell line INS‐1 were pre‐treated with the indicated concentration of LNT for 30 min. and then incubated for 24 hrs with or without 0.5 mM STZ. We found that STZ treatment causes apoptosis of INS‐1 cells by enhancement of intracellular reactive oxygen species (ROS) accumulation, inducible nitric oxide synthase (iNOS) expression and nitric oxide release and activation of the c‐jun N‐terminal kinase (JNK) and p38 mitogen‐activated protein kinase (MAPK) signalling pathways. However, LNT significantly increased cell viability and effectively attenuated STZ‐induced ROS production, iNOS expression and nitric oxide release and the activation of JNK and p38 MAPK in a dose‐dependent manner in vitro. Moreover, LNT dose‐dependently prevented STZ‐induced inhibition of insulin synthesis by blocking the activation of nuclear factor kappa beta and increasing the level of Pdx‐1 in INS‐1 cells. Together these findings suggest that LNT could protect against pancreatic β‐cell apoptosis and dysfunction caused by STZ and therefore may be a potential pharmacological agent for preventing pancreatic β‐cell damage caused by oxidative stress associated with diabetes.

Hexarelin Protects Rodent Pancreatic Β-Cells Function from Cytotoxic Effects of Streptozotocin Involving Mitochondrial Signalling Pathways In Vivo and In Vitro.

Mitochondrial functions are crucial for pancreatic β-cell survival and glucose-induced insulin secretion. Hexarelin (Hex) is a synthetic small peptide ghrelin analogue, which has been shown to protect cardiomyocytes from the ischemia-reperfusion process. In this study, we used in vitro and in vivo models of streptozotocin (STZ)-induced β-cell damage to study the protective effect of Hex and the associated mechanisms. We found that STZ produced a cytotoxic effect in a dose- and time-dependent manner in MIN6 cells (a mouse β-cell line). Hex (1.0 μM) decreased the STZ-induced damage in β-cells. Rhodamine 123 assay and superoxide DHE production assay revealed that Hex ameliorated STZ-induced mitochondrial damage and excessive superoxide activity in β-cells. In addition, Hex significantly reduced STZ-induced expression of cleaved Caspases-3, Caspases-9 and the ratio of pro-apoptotic protein Bax to anti-apoptotic protein Bcl-2 in MIN6 cells. We further examined the in vivo effect of Hex in a rat model of type 1 diabetes induced by STZ injection. Hex ameliorated STZ-induced decrease in plasma insulin and protected the structure of islets from STZ-induced disruption. Hex also ameliorated STZ-induced expression of cleaved Caspase-9 and the Bax in β-cells. In conclusion, our data indicate that Hex is able to protects β-cell mass from STZ-caused cytotoxic effects involving mitochondrial pathways in vitro and in vivo. Hex may serve as a potential protective agent for the management of diabetes.

Anti-Hyperglycemic Effect of Tecomella undulata Extract by Ameliorating Pancreatic Dysfunction in Streptozotocin Induced Diabetic Albino Rats

Article history: Objective: The primary objective of the present investigation is to evaluate the antihyperglycemic activity of the 50% ethanolic extract of the Tecomella undulata Seem. (TUE) bark in the streptozotocin induced diabetic rats. Methods: Single intraperitoneal injection (i.p.) of streptozotocin (60 mg/kg body weight) was used for induction of diabetes is wistar albino rats. The induction of diabetes was confirmed after 3 days after STZ injection and rats with fasting blood glucose levels greater than 200 mg/dl were considered to be diabetic used in the experiment. TUE at a once a daily dose of 250 mg/kg, and 500 mg/kg, p.o. along with glibenclamide 600µg/kg, p.o. was also given for 21 days. On the 21 st day rats from all the groups fasted overnight and the blood was collected from the puncturing the retro orbit of the eye under mild ether anesthetic condition. The oral acute toxicity studies did not show any toxic effect till the dose at 2000 mg/kg. Results: The statistical data indicated that the different doses of the TUE significantly decreases the level of fasting blood glucose, total cholesterol, triglycerides, low density lipoprotein and increase high density lipoprotein in STZ induced diabetic rats. This result indicated that T. undulata extract can protect pancreatic β- cells from STZ-induced damage which is confirmed by the results of the histopathological examination of the pancreas. Conclusion: Our investigation has clearly indicated that the bark extract of Tecomella undulata Seem. showed remarkable antihyperglycemic activity due to its possible systematic effect involving in the pancreatic and extra pancreatic mechanism. Moreover, the antihyperlipidemic activity was exerted possible by lowering the higher level of lipid profile.

Hypoglycemic effect of deoxynojirimycin–polysaccharide on high fat diet and streptozotocin-induced diabetic mice via regulation of hepatic glucose metabolism

Type 2 diabetes mellitus (T2DM) is currently considered a worldwide epidemic and finding effective therapeutic strategies against this disease is highly important. A deoxynojirimycin–polysaccharide mixture (DPM) has previously been shown to exert hypoglycemic effects on alloxan- or streptozotocin (STZ)-induced diabetic mice. The purpose of the present study was to evaluate the therapeutic effects and underlying mechanism(s) of DPM on T2DM induced by high fat diet following low-dose STZ treatment in mice. After daily oral treatment of diabetic mice with DPM (150mg/kgb.w.) for 90d, significant decline in blood glucose, pyruvate, triglyceride (TG), aspartate transaminase (AST), alanine transaminase (ALT), creatinine (Cr), lipid peroxide (LPO) and malondialdehyde (MDA) levels as well as evident increases in high density lipoprotein (HDL-c) and hepatic glycogen concentrations were observed. In the first stage, in which DPM was administered for 60d, blood insulin levels did not undergo significant change but a significant decrease in the HOMA-IR index was detected. By contrast, the HOMA-IR index increased significantly in T2MD controls. In the second stage, in which DPM treatment was continued for another 30d, insulin levels significantly increased in DPM-treated mice in comparison with T2DM controls. These results indicate that insulin resistance in the pre-diabetic period and the dysfunction of pancreatic β-cells are ameliorated by DPM treatment. DPM also down-regulated protein levels of insulin receptor (IR) and gluconeogenic enzymes (pyruvate carboxylase, fructose-1, 6-bisphosphatase, phosphoenolpyruvate carboxykinase and glucose-6-phosphatase) in peripheral tissues (liver and/or muscle), but enhanced the expressions of insulin in pancreas, lipoprotein lipase (LPL) and glycolysis enzymes (glucokinase, phosphofructokinase, private kinase and pyruvate decarboxylase E1) in the liver. Furthermore, deoxynojirimycin (DNJ) and polysaccharide (P) were found to increase proliferation of hepatic LO-2 cells and scavenging of radicals in vitro. These results support the results of our biochemical analyses and underscore possible mechanisms underlying the protective effects of DPM on STZ-induced damage to the pancreas and the liver. Taken together, our findings suggest that DPM may be developed as an antihyperglycemic agent for the treatment of diabetes mellitus.

Cooperative Anti-Diabetic Effects of Deoxynojirimycin-Polysaccharide by Inhibiting Glucose Absorption and Modulating Glucose Metabolism in Streptozotocin-Induced Diabetic Mice

We had previously shown that deoxynojirimycin-polysaccharide mixture (DPM) not only decreased blood glucose but also reversed the damage to pancreatic β-cells in diabetic mice, and that the anti-hyperglycemic efficacy of this combination was better than that of 1-deoxynojirimycin (DNJ) or polysachharide alone. However, the mechanisms behind these effects were not fully understood. The present study aimed to evaluate the therapeutic effects of DPM on streptozotocin (STZ)-induced diabetic symptoms and their potential mechanisms. Diabetic mice were treated with DPM (150 mg/kg body weight) for 90 days and continued to be fed without DPM for an additional 30 days. Strikingly, decrease of blood glucose levels was observed in all DPM treated diabetic mice, which persisted 30 days after cessation of DPM administration. Significant decrease of glycosylated hemoglobin and hepatic pyruvate concentrations, along with marked increase of serum insulin and hepatic glycogen levels were detected in DPM treated diabetic mice. Results of a labeled 13C6-glucose uptake assay indicated that DPM can restrain glucose absorption. Additionally, DPM down-regulated the mRNA and protein expression of jejunal Na+/glucose cotransporter, Na+/K+-ATPase and glucose transporter 2, and enhanced the activities as well as mRNA and protein levels of hepatic glycolysis enzymes (glucokinase, phosphofructokinase, private kinase and pyruvate decarboxylas E1). Activity and expression of hepatic gluconeogenesis enzymes (phosphoenolpyruvate carboxykinase and glucose-6-phosphatase) were also found to be attenuated in diabetic mice treated with DPM. Purified enzyme activity assays verified that the increased activities of glucose glycolysis enzymes resulted not from their direct activation, but from the relative increase in protein expression. Importantly, our histopathological observations support the results of our biochemical analyses and validate the protective effects of DPM on STZ-induced damage to the pancreas. Thus, DPM has significant potential as a therapeutic agent against diabetes.

Effect of Potent Ethyl Acetate Fraction ofStereospermum suaveolensExtract in Streptozotocin-Induced Diabetic Rats

To evaluate the antihyperglycemic effect of ethyl acetate fraction of ethanol extract of Stereospermum suaveolens in streptozotocin-(STZ-) induced diabetic rats by acute and subacute models. In this paper, various fractions of ethanol extract of Stereospermum suaveolens were prepared and their effects on blood glucose levels in STZ-induced diabetic rats were studied after a single oral administration (200 mg/kg). Administration of the ethyl acetate fraction at 200 mg/kg once daily for 14 days to STZ-induced diabetic rats was also carried out. The parameters such as the fasting blood glucose, hepatic glycogen content, and pancreatic antioxidant levels were monitored. In the acute study, the ethyl acetate fraction is the most potent in reducing the fasting serum glucose levels of the STZ-induced diabetic rats. The 14-day repeated oral administration of the ethyl acetate fraction significantly reduced the fasting blood glucose and pancreatic TBARS level and significantly increased the liver glycogen, pancreatic superoxide dismutase, and catalase activities as well as reduced glutathione levels. The histopathological studies during the subacute treatment have been shown to ameliorate the STZ-induced histological damage of pancreas. This paper concludes that the ethyl acetate fraction from ethanol extract of Stereospermum suaveolens possesses potent antihyperglycemic and antioxidant properties, thereby substantiating the use of plant in the indigenous system of medicine.

Polyphenols-rich Vernonia amygdalina shows anti-diabetic effects in streptozotocin-induced diabetic rats

This study aims to investigate the hypoglycemic properties of Vernonia amygdalina Del. (VA) and its possible mechanisms of action in a single-dose STZ induced diabetic rat model. A dose-response study was conducted to determine optimum dose for the hypoglycemic effect of VA in STZ-induced diabetic rats. The optimum dose (400 mg/kg) was used throughout the 28-day chronic study. Body weight, food and water intakes of the rats were monitored daily. Fasting blood serum, pancreas, liver and soleus muscle were collected for biochemical analyses. Chemical composition of VA was analysed using HPLC and LC-ESI-MS. The study reveals that ethanolic extract of VA contains high level of polyphenols mainly 1,5-dicaffeoyl-quinic acid, dicaffeoyl-quinic acid, chlorogenic acid and luteolin-7-O-glucoside. In an oral glucose tolerance test, 400 mg/kg VA exhibited a significant improvement in glucose tolerance of the STZ-induced diabetic rats. 28-day treatment with 400 mg/kg VA resulted in 32.1% decrease in fasting blood glucose compared to diabetic control. VA also caused significant decrease (18.2% and 41%) in triglyceride and total cholesterol level. Besides, VA showed protective effect over pancreatic β-cells against STZ-induced damage, causing a slight increase in insulin level compared to diabetic control. VA administration also showed positive regulation of the antioxidant system, both enzymatic and non-enzymatic. Furthermore, VA was found to increase expression of GLUT 4 (24%) in rat skeletal muscle. Further tissue fractionation revealed that it can increase the GLUT 4 translocation (35.7%) to plasma membrane as well, suggesting that VA may stimulate skeletal muscle's glucose uptake. This observation is in line with the restoration in skeletal muscle glycogenesis of VA-treated group. However, no alteration was observed in GLUT 1 expression. In addition, VA also suppressed (40% inhibition) one of the key hepatic gluconeogenic enzymes, glucose-6-phosphatase (G6Pase). VA possesses antihyperglycemic effect, most probably through increasing GLUT 4 translocation and inhibiting hepatic G6Pase. The polyphenols in the extract may be the candidates that are responsible for the above-mentioned biological activities.

Antihyperglycaemic and protective effects of flavonoids on streptozotocin–induced diabetic rats

The antihyperglycaemic effect of eight standard flavonoids, previously identified in the ethanol extract of the claimed antidiabetic plant Genista tenera, was evaluated on streptozotocin (STZ)-induced diabetic Wistar rats. The aglycones apigenin, chrysoeriol and genistein, the monoglucosides apigenin 7-O-glucoside, luteolin 7-O-glucoside and genistein 7-O-glucoside and the diglycosides rutin and luteolin 7,3'-di-O-glucoside were administered i.p. for 7 days (4 mg/kg b.w./day). The protective effect of these compounds over liver and kidneys of STZ-diabetic models was also evaluated by the determination of seric AST, ALT and urea levels. After 7 days of treatment, apigenin, chrysoeriol and genistein significantly lowered the blood glucose levels of diabetic animals; this effect was more pronounced (P < 0.01) in the oral glucose tolerance test. Glucose tolerance was also significantly improved in the rutin (P < 0.01) and in the genistein 7-O-glucoside (P < 0.05) treated groups. In addition, almost all the tested compounds effectively protected the liver and kidneys against STZ-induced damage in rats.

The influence of rutin on the extracellular matrix in streptozotocin-induced diabetic rat kidney

We previously reported that rutin administration to streptozotocin (STZ)-induced diabetic rats decreased plasma glucose and increased plasma insulin levels. In this study, we have examined the role of rutin on matrix remodelling in the kidney of STZ-induced diabetic rats. STZ was administered intraperitoneally (50 mg kg(-1)) to male albino Wistar rats to induce experimental diabetes. Rutin (100 mg kg(-1)) was orally administered to normal and STZ-induced diabetic rats for a period of 45 days and its influence on the content of hydroxyproline and collagen and on the activity of matrix metalloproteinases (MMPs) were studied. We have also studied the levels of tissue inhibitors of metalloproteinases (TIMPs) in the kidney. STZ-induced diabetic control rats showed increased content of hydroxyproline and collagen, decreased activity of MMPs and increased levels of TIMPs in the kidney. These changes were positively modulated by rutin treatment in STZ-induced diabetic rats, thereby protecting the kidney. In normal rats treated with rutin, none of the parameters studied were significantly altered. From the results obtained, we could conclude that rutin influences MMPs and effectively protects kidney against STZ-induced damage in rats. The effects observed are due to the reduction of plasma glucose levels by rutin.

KIOM-4 protects RINm5F pancreatic β-Cells against streptozotocin induced oxidative stress in vitro

The effect of KIOM-4, a combination of four plant extracts was assessed on streptozotocin (STZ) treated rat insulinoma (RIN5mF) cells in vitro. KIOM-4 scavenged the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and intracellular reactive oxygen species (ROS) induced by STZ. KIOM-4 prevented the STZ-induced DNA damage, which is detected using comet assay, Western blot and lipid peroxidation assays. KIOM-4 inhibited the STZ induced apoptosis, therefore protecting from cell death. Additionally, KIOM-4 induced the activation of catalase and extracellular regulated kinase (ERK). These results suggest that KIOM-4 protects RINm5F cells via radical scavenging activity, the activation of catalase and ERK against STZ induced oxidative stress.

A quantitative study of sodium tungstate protective effect on pancreatic beta cells in streptozotocin-induced diabetic rats

Diabetes is a major public health problem. Development of new therapies that are able to improve glycemia management, cure diabetes, and can even protect from it, are of great interest. This study investigated the protective effect of sodium tungstate against STZ-induced beta-cell damages by means of stereological methods. Sixty rats were divided into six groups: control (C), tungstate-treated control (TC), STZ-induced diabetic (D), STZ-induced diabetic rats were treated by sodium tungstate from 1 week before STZ injection (TDB), food-restricted diabetic (FRD), and diabetic rats treated with sodium tungstate 1 week after STZ administration (TDA). Stereological estimation of pancreas volume, islets volume density, volume-weighted mean islets volume and mass of beta cells, islets, and pancreas and total number of islets were done. Islets volume density, volume-weighted mean islets volume, and mass of beta cells, islets, and pancreas of TDB group was significantly higher than D, FRD and TDA groups ( P < 0.001) and was comparable to controls (C and TC groups). Total number of islets, pancreas wet weight and volume did not show any significant changes between these groups ( P > 0.05). Results suggested that sodium tungstate preserves pancreatic beta cells from STZ-induced damages and diabetes induction in rats.

Heterogenous changes in neuropeptide Y, norepinephrine and epinephrine concentrations in the hearts of diabetic rats

The changes in concentrations of neuropeptide Y (NPY), norepinephrine and epinephrine were investigated in the rat hearts 1, 2, 4, 6, 9 and 12 months after administration of streptozotocin (STZ; 65 mg/kg i.v.). About 30% of diabetic animals displayed symptoms of partial spontaneous recovery, i.e. decreasing blood glucose levels and increasing insulin concentrations in the plasma and pancreas. NPY concentrations in the atria of diabetic rats did not differ from those in age-matched control rats 1, 2, 4, 6 months in the right atria and even 9 months after STZ in the left atria. However, uncompensated diabetes led to a significant decrease in NPY levels 9 and 12 months after STZ administration in the right and left atria, respectively. In the ventricles, NPY concentrations were significantly decreased 6 months after the onset of diabetes. Interestingly, partial spontaneous recovery of diabetes was associated with increased NPY levels in the atria. Myocardial norepinephrine concentrations increased 1 month after STZ and then declined reaching ∼ 60% of the respective control values 12 months after the onset of the disease. Partial spontaneous recovery of diabetes had no effect on norepinephrine concentrations. Myocardial epinephrine concentrations did not differ from those found in controls till month 9 of the disease and they became significantly lower at month 12. Partial recovery of diabetes resulted in epinephrine concentrations not differing from the control values at month 12 of diabetes. Regarding to preferential localization of norepinephrine in the sympathetic postganglionic fibers and that of NPY also in intrinsic ganglion neurons, intrinsic neuronal circuits seem to be less susceptible to STZ-induced damage than extrinsic nerves and they might be able to recover after amelioration of diabetes.

Superoxide dismutase reduces islet microvascular injury induced by streptozotocin in the rat.

Intravenous administration of streptozotocin (STZ) leads to permanent diabetes mellitus in rats. We investigated the possible role of islet microcirculatory changes and free radical formation in this animal model. In vivo fluorescence microscopy was performed for 4 h after administration of STZ. Vascular permeability, capillary blood flow, and endothelial leukocyte adhesion were measured in endocrine and exocrine pancreatic tissue. The earliest microcirculatory event was an increase in vascular permeability in pancreatic islets, with a peak 1 h after STZ administration. The difference between islet and exocrine tissue light intensity was +15.8 +/- 5.6% at t = 60 min. Islet blood flow velocity significantly decreased after 3 h, whereas blood flow in the exocrine pancreas was not affected. Complete stasis of islet blood flow was observed only in rats receiving STZ. Neither increased leukocyte adhesion to islet vascular endothelium nor ischemia-reperfusion phenomena were observed. Prophylactic administration of the radical scavenger superoxide dismutase prevented STZ-induced damage to the islet microcirculation in the initial phase of this model. We conclude that STZ leads to severe microcirculatory disturbances within pancreatic islets in rats. Apparently, these changes are mediated at least in part by free oxygen radicals.

Involvement of spontaneous nitric oxide production in the diabetogenic action of streptozotocin.

The exact mechanism by which streptozotocin (STZ) destroys pancreatic beta-cells has not been determined. Recently, nitric oxide was identified as a potent islet toxic compound. However, nitric oxide production by STZ has not been proved yet. In the present studies, we determined whether STZ spontaneously produces nitrite, measured by a spectrophotometric technique based on the Griess reaction. STZ, dissolved in the pH 4.0 citrate buffer, produced significantly more nitrite than that dissolved in pH 7.4. After incubation at 37 degrees C for 120 min, nitrite concentrations were significantly decreased only in STZ dissolved in the pH 7.4 citrate buffer. Following 120 min incubation at 37 degrees C, STZ dissolved at pH 7.4 failed to cause significant islet damage in rats previously deprived of food for 18 h. The present studies demonstrated that STZ may spontaneously produce nitrite in vitro. Nitric oxide production may, at least in part, be responsible for the STZ-induced damage of pancreatic beta-cells of the rodents.

Mechanisms of STZ-induced cell damage: the role in cell survival, mutagenesis and chromosomal alterations

Mechanisms of STZ-induced cell damage: the role in cell survival, mutagenesis and chromosomal alterations