Effect Of Dachaihu Decoction On Oxidative Stress Induced Beta-cell Injury Of Islet In Rats With Type 2 Diabetes

Objective To observe the changes of oxidative stress response factors ROS,MDA,SOD in serum and the expression of PDX-1 and MafA mRNA in pancreatic tissue before and after treatment with Da Chaihu decoction in rats with type 2 diabetes mellitus.To explore the effect of Dachaihu Decoction on islet of type 2 diabetic rats.Cell protection mechanism provides experimental basis for clinical application.Methods Fifty male Sprague-Dawley rats were fed for 3 days,10 were randomly selected as the normal control group,and the other 40 rats were fed with high-sugar and high-fat diet for 4 weeks.The intraperitoneal injection of streptozotocin(STZ)replicated type2 diabetes.Mouse model.After modeling,they were randomly divided into Dachaihu Decoction group,simvastatin group,metformin group and model control group.The normal control group was fed with normal feed,and the model group was fed with high-sugar and high-fat diet.The model control group and the normal control group were given the same amount of normal saline,the Dachaihu Decoction group was given Da Chaihu Decoction10.1g/kg/d,and the simvastatin group was given simvastatin suspension 20mg/kg/d.Metformin group was given 300 mg/kg/d of metformin suspension.After 8 weeks of gavage,fasting for 12 hours(not allowed for water),fasting blood glucose(FBG)was detected the next morning,and abdominal aorta was taken after anesthesia.Pancreatic tissue;detection related indicators: detection of glycosylated hemoglobin(Hb A1c),total cholesterol(TC),triglyceride(TG),free fatty acid(FFA);ELISA detection of ROS,MDA,SOD content,calculate antioxidant ratio SOD /MDA;Real-time quantitative PCR(RT-qPCR)was used to detect the expression of PDX-1 and MafA mRNA in pancreatic tissue;HE staining was used to observe the pathological changes of pancreatic tissue.Results 1.The results of blood glucose and blood lipid biochemical indicators: compared with the normal control group,the model group FBG,HbA1 c,TC,TG,FFA significantly increased(P <0.01);compared with the model group,simvastatin group,Dachaihu group FBG,HbA1 c,TC,TG and FFA were significantly lower in metformin group(P<0.01).Compared with simvastatin group,TG decreased in Dachaihutang group(P<0.05),and FBG,HbA1 c,TC and FFA decreased significantly(P<0.05).P<0.01);Compared with the metformin group,the FFA of the Dachaihu Decoction group was significantly decreased4(P<0.01),and the difference of FBG,HbA1 c,TC and TG was not statistically significant(P>0.05).2.ELISA test results: The results of ELISA showed that compared with the normal control group,the ROS,SOD and MDA in the model group were significantly increased(P<0.01).Compared with the model group,simvastatin group,Dachaihutang group,metformin group FBG,HbA1 c,TC TG and FFA were significantly decreased(P<0.01).Compared with simvastatin group,TG decreased in Dachaihutang group(P<0.05),FBG,HbA1 c and TC decreased significantly(P<0.01),FFA decreased but there was no difference.Statistical significance(P>0.05);Compared with metformin group,there was no significant difference in FBG,HbA1 c,TC,TG and FFA between Dachaihu Decoction group(P>0.05).3.RT-PCR test results :The results of RT-PCR showed that compared with the normal control group,the expression of PDX-1 and MaFA mRNA in the pancreatic tissue of the model group was significantly decreased(P<0.01).Compared with the model group,the simvastatin group,The expression of PDX-1 mRNA in pancreatic tissue of Dachaihutang group and metformin group was significantly increased(P<0.01),and the expression of MaFA mRNA in pancreatic tissue of metformin group was increased(P<0.05).Simvastatin group and Dachaihu decoction group The expression of MaFA mRNA in rat pancreas was significantly increased(P<0.01).4.Pathological morphology of islet tissue: There are a large number of normal islets in the pancreas of normal control group,the tissue structure is clear,the cells are arranged neatly,and the acinar lobule is intact.Only a small amount of residual fibrotic islets are found in the pancreas of the model group,and the structure is disordered.The acinar lobular contour destroys the inflammatory cell infiltration;the simvastatin group has scattered scattered islets in the pancreas,the acinar lobular structure is relatively intact,and a small amount of inflammatory cell infiltration is observed;in the metformin group,there are scattered new islets in the pancreas Distribution,acinar lobular structure is relatively complete,a small amount of inflammatory cell infiltration can be seen;Dashuihu decoction group has obvious distribution of new islets in the pancreas,and the number of islets is significantly increased compared with the model group,occasionally a small amount of inflammatory cell infiltration.Conclusion1.It is confirmed that oxidative stress-mediated pancreatic ?-cell damage is an important olecular mechanism in the early stage of type 2 diabetes.2.The main mechanism of action of “Kuyu Qingre” and its representative Fang Dachaihu Decoction in the treatment of type 2 diabetes is to inhibit oxidative stress and achieve the effect of protecting islet ? cells.