| Objective:The common pathological factor of diabetes caused by various complications is the increased oxidative stress, thus anti-oxidative stress is the important candidate auxiliary method to treatment diabetes complications. In this experiment we established diabetes rats and to treated it by Xin mailong injection,and then systematically detected the related indicators of oxidative stress of rats in different groups, to discuss whether the Xin mailong injection has effects to alleviate the oxidative stress caused by diabetes. Through this study we hope to clear the effects of Xin mailong injection in the role of the anti-oxidative stress, to expanding understanding of pharmacological effects of periplaneta americana extract, to providing the theoretical basis for new drug research and development in the future.Content : Fist, we established the experimental diabetic rats model which simulated the type 2 diabetes of human, then systematically detectd diabetes related indictors to confirm that the model is established successfully. On this basis,we given Xin mailing(XML) injection to diabetes rats, systematically detected the related indicators of oxidative stress in serum and brains of rats, which including:the content of lipid oxidation product Malondialdehyde(MDA)、the changes of Total antioxidant capacity(TAOC)、the activity of Catalase(CAT) and the changes of protein and DNA oxidative stress injury in brain. By comparing the changes of these indicators, we discuss whether the Xin mailong injection play to improve the oxidative stress caused by diabetes.Method :Establishing a rat model of type 2 diabetes mellitus by feeding high-fat diet combined with a smll does streptozotocin(STZ) injection. In the stage of building the model of diabetes, the adult male Sprague-Dawley(SD) ratswere randomly assigned to two groups(the XML injection before): normal control group(control, n=10 normal diet); model group( DM n=30,feeding fat diet),weekly weighing rat weight. After feeding high fat feed 8 weeks, rats in DM group were given STZ 25mg/kg by intraperitoneal injection. Continue feeding four weeks to complete the model, we systically detected the related indicators of type 2 diabetes,including the changes of weight, changes in blood glucose, serum triglycrides levels, plasma cholesterol levels and insulin sensitivity etc, to determine that the model is established successfully. Based on this rat model, we further researched the possible role of XML in anti oxidative stress by drug intervening treatment as following: diabetic rats were randomly divided into XML treatment group(XML,n=10, feeding fat diet +XML injection)、diabetes group(DM,n=18) and control group(control,n=10), then to continue feeding 25 days. During this period,diabetes XML treatment rats were injected i.p according to the set of daily doses( 9mg/kg), keep dynamic monitoring the changes of general physiological indexes, such as coat colour, activity, drinking water, food and urine etc. Then we obtained the blood and brain tissue samples respectively. To compare the oxidative stress condition in different groups, we detected the lipid oxidation product generation, changes of oxidation resistance and the changes of hydrogen peroxide enzyme by using specific kit, detected the protein oxidative damage in the brain by Western blotting and detected DNA oxidative damage in the brain by immunohistochemical staining.Results:1. Type 2 diabetic rats mode established by feeding high fat diet and one-time intraperitoneal injection of low-dose STZ can mimic typical diabetes symptoms in human being.1.1 The changes of rats’ weight: weight of rats DM groups was obviously higher than that of control group, significantly higher than the growth rate of control groups. Pre-diabetes obesity appeared.1.2 Compared with the control group, rats of DM group insulin sensitivity decreased indicating insulin resistance occured.1.3 The changes of urine volume and quantity of drinking water of rats: urine,water quantity of rats in DM groups was obviously higher than that of the control group(p<0.05). Urine has no obvious improve after XML injection treatment.1.4 The determination of blood glucose, triglyceride, and cholesterol levels:DM group rats blood glucose, triglyceride, and cholesterol significantly higher than the control group. Hyperglycemia and hyperlipidemia appeared.1.5 Observe the appearance of liver:the liver of control group is dark red、normal shape、smooth. The size of the liver of DM group is significantly greater than that in control group. The liver is creamy yellow, greasy feeling, liver capsule tension, and its surface have no gloss. The size of the liver of XML group somewhere in between, the liver color lighter than control group but compared with DM group is significantly improved.2 、 XML plays a role in alleviating oxidative stress caused by diabetes conditions.2.1 The determination of the MDA of blood plasma 、 cerebrum and hippocampus:XML treatment can decrease lipid oxidated product MDA value which is markedly increased in DM group(p<0.05) in plasma and brain. This indicates that XML injection has the certain effect on anti lipid oxidation process caused by diabetes condition.2.2 Determination of plasma antioxidant capacity: antioxidant capacity of DM group rats is higher than the control and XML groups(p<0.05).This indicates that the body may have a compensatory period in oxidative stress. Total antioxidant capacity increasing in the compensatory phase, XML injection can delay the process of compensatory to decompensation of the body oxidative stress.2.3 Determination of cerebral cortex’s and hippocampus’ s antioxidant capacity: the antioxidant capacity of DM group rats reduced(p<0.05), the antioxidant capacity is higher than the DM group after treatment with XML injection(p<0.05).Tip: XML injection has certain effect on oxidative stress of brain tissue, its can improve the antioxidant capacity of brain tissue.2.4 Determination of the catalase in cerebral cortex and hippocampus:the catalase activity of DM group rats is higher than control group and XML group(p<0.05). This indicates that XML injection has certain protection effect on antioxidase.2.5 Protein and DNA oxidative damage detection: carbonylation modified protein increased significantly in brain of diabetes rats, after deal with XML, a slight downward trend was obsered; DNA oxidation increased in rats of DM group, however, after treatment by XML injection, DNA oxidative product8-oxo-d G positive staining reduced. This result indicates that XML injection can reduce the brain protein and DNA oxidative damage product.Conclusions:1 、 The method of long time high-fat diet raising and small does STZ treatment to rats can establish rat model which simulated type 2 diabetes in human in some respects.2、XML injection could delay the process of oxidative stress, improve the body antioxidant capacity and reduce the protein and DNA oxidative damage in the brain by eliminating superoxide and protect antioxidant. It can inhibit, delay and improve oxidative stress caused by diabetes. |
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