Shizidaipingfang Ameliorate Insulin Esistance In Insulin-Resistant HepG2 Cells And Its Influence On The Content Of The Liver Glycogen,HK,PK,GLUT-2

Objectives To investigate the protection of Shizidaipingfang of the high insulin induced insulin protection against HepG2 cells, and to explore its mechanism of action by detecting the glucose consumption, the proliferation, the content of hepatic glycogen, HK activity,PK activity and GLUT-2.Methods 1. The model of insulin resistance was established in HepG2 cells by induction of high concentration insulin in vitro, the proliferation of HepG2 cells was detected by MTT, the glucose consumption of cells was detected by GOD-POD, and then screened out the best modeling concentration and the time of modeling. 2. The glucose consumption and the proliferation of IR-HepG2 cell model were detected in Shizidaipingfang, and then screened out the optimal concentration of Shizidaipingfang. 3. The effects of kinase(HK)activity, pyruvate kinase(PK) activity and the content of hepatic glycogen of IR-HepG2 cell model were detected in Shizidaipingfang, at the same time, The expression of glucose transporter 2(GLUT-2) were detected by Western-blot method in IR-HepG2 cells.Results 1. No effect of Insulin, which concentration was 1×10-5 ~ 1×10-10 mol?L-1, on HepG2 cells proliferation was detected with MTT method. Through the difference of glucose consumption of the cells in each group, the optimal model concentration was 1×10-7mol?L-1, and the best model time was 36 h. 2. No effect of Shizidaipingfang in each concentration on IR-HepG2 cells proliferation was detected with MTT method(P>0.05),and the survival rate of the cells in each group were more than 90%. Compared with model group, there were a significant increase in the amount of glucose consumption of IRHepG2 cell model in different concentration of Shizidaipingfang(P<0.01), Among them,its concentration was the most obvious in 100μg?ml-1. 3. Compared with the normal group,the glycogen content, HK activity and PK activity were significantly decreased in the model group(P<0.05). Compared with model group, the glycogen content, HK activity and PK activity were significantly risen in the Shizidaipingfang group, which had statistical significance(P<0.05), but compared with normal group, which had not obvious difference(P>0.05). 4. Western blot results show, Model group compared with the normal group, with significant difference(P<0.05). Compared with model group, shizidaipingfang can significantly improve the level of protein expression of GLUT-2(P<0.05).Conclusions 1. Shizdaipingfang could increase the glucose consumption of IR-HepG2 cells, and had no effect on cell proliferation, so suggested that Shizidaipingfang can improve HepG2 cell insulin resistance. 2. Shizidaipingfang could increase the content of hepatic glycogen, increase the activity of PK and the activity of HK, and upregulated GLUT-2 protein expression levels. 3. Shizidaipingfang improved the mechanism of insulin resistance in HepG2 cells may be achieved by increased the activity of HK, PK activity,increased the content of hepatic glycogen and upregulated of GLUT-2 protein expression level, and ultimately increased the use of glucose in cells.