Study On The Intervention Effect And Mechanism Of Banxia Xiexin Decoction On Insulin Resistance Of Skeletal Muscle Induced By 3-deoxyglucosone

Part I Study on the intervention effect and mechanism of Banxia Xiexin Decoction on skeletal muscle of rats with impaired glucose regulation induced by 3-deoxyglucosoneObjective:To study the effect of Banxia Xiexin Decoction on the skeletal muscle of IGR rats induced by 3DG.Methods:SD rats were randomly divided into three groups according to their body weight:blank control group,model group(3DG,50 mg/kg),Banxia Xiexin Decoction group(3DG,50mg/kg+Banxia Xiexin Decoction group,10 g/kg),10 rats in each group,once a day by gavage for two weeks.After 2 weeks of administration,fasting blood glucose and OGTT were measured;serum of rats was taken and insulin level was measured by ELISA kit;membrane protein and total protein of skeletal muscle cells were extracted,and protein expression of GLUT4,IR-? and p-IR-? on the membrane of skeletal muscle cells were measured by western blot.Results:The results of fasting blood glucose experiment showed that:compared with 3DG model group,Banxia Xiexin Decoction significantly reduced fasting blood glucose level(P<0.05).The results of oral glucose tolerance test showed that:compared with the 3DG model group,Banxia Xiexin Decoction could significantly reduce the blood glucose level of rats at 30 min and 60 min time points(P<0.05).The results of ELISA showed that:compared with 3DG model group,Banxia Xiexin Decoction could significantly increase the serum insulin level of rats(P<0.05).Western blot showed that Banxia Xiexin Decoction could significantly up regulate GLUT4,IR-? and the level of skeletal muscle(P<0.05).Conclusion:Banxia Xiexin Decoction can down regulate the fasting blood glucose level of IGR rats induced by 3DG,up regulate the serum insulin level,up regulate the GLUT4 translocation and IR-? protein expression of skeletal muscle cells,promote the glucose transport in skeletal muscle,so as to improve the impaired glucose tolerance of rats induced by 3DG.Part ? Study on the effect and mechanism of the serum containing Banxia Xiexin Decoction on the impaired glucose regulation of L6 cells induced by 3-deoxyglucosoneObjective:To investigate the effect of Banxia Xiexin Decoction on IGR of L6 cells induced by 3DG.Based on the PI3K-Akt signal pathway,the molecular mechanism of Banxia Xiexin Decoction regulating insulin resistance by interfering with GLUT4 translocation of skeletal muscle was studied.Methods:To explore the concentration and time of inhibition of glucose uptake by L6 cells induced by 3DG.To explore the best concentration of Banxia Xiexin Decoction containing serum to improve the impaired glucose uptake of L6 cells induced by 3DG.The cell viability and 2-NBDG fluorescence intensity were measured by MTT method to evaluate the cell glucose uptake.Western blot was used to detect the expression of GLUT4 and PI3K-Akt signal pathway key proteins on L6 cell membrane.Results:Compared with the insulin control group,the glucose uptake ability of L6 cells decreased significantly 24 hours after 3DG intervention,especially when the concentration of 3DG reached 300 ng/ml(P<0.05).Compared with the model group,when the serum containing Banxia Xiexin Decoction was 5%,the glucose intake increased significantly(P<0.01)Compared with 3DG group,the expression of GLUT4 protein in Banxia Xiexin Decoction group and 3DG+aminoguanidine group increased significantly(P<0.001).It is suggested that Banxia Xiexin Decoction can improve glucose uptake of L6 cells with insulin resistance through translocation of GLUT4 membrane.Compared with the blank control group,the expression of p-IR protein in the 3DG group decreased significantly(P<0.001);compared with the blank control group,the expression of p-Akt and protein in the 3DG group decreased(P<0.001);compared with the 3DG group,the expression of p-IR protein increased significantly(P<0.05).Compared with 3DG group,the expression of p-Akt and PI3K-p110 protein in Banxiaxiexin Decoction group increased significantly(P<0.001).Conclusion:Banxia Xiexin Decoction may up regulate GLUT4 translocation of skeletal muscle cells through PI3K-Akt signal pathway,improve the glucose uptake function of L6 cells induced by 3DG,and then intervene the impairment of insulin resistance of skeletal muscle cells.