| Objective:To study the effect of Dahuang Huanglian Xiexin Decoction on PI3K/Akt signaling pathway in ir-hepg2（insulin resistance human hepatoblastoma cell）model induced by high concentration of insulin Objective to investigate the effect of diabetes mellitus（T2DM）on glucose and lipid metabolism and the mechanism of whether it alleviates IR.Methods:（1）Hep G2 cells were cultured,resuscitated and subcultured;（2）high concentration insulin was used to induce the model in vitro,and different insulin concentration gradients were set to treat Hep G2 cells,and the best time to establish the model was determined by CCK-8 method when the insulin concentration was 10-7mg/L and the treatment time was 48h;（3）different concentration gradients of Dahuang Huanglian Xiexin Decoction and Jinqi Decoction were set The IC50 of Dahuang Huanglian Xiexin Decoction was 1.2*10-5mg/ml,and that of Jinqi Jiangtang Tablet was 2*10-4mg/ml.according to the results,Dahuang Huanglian Xiexin Decoction group was divided into high-dose group,medium dose group and low-dose group.The drug concentration of high-dose group was 1.2*10-5mg/ml,medium dose group was 6*10-6mg/ml,and low-dose group was 3*10-6mg/ml The optimal drug treatment time was 24 h at the concentration of1*10-5 mg/ml;（4）the levels of triglyceride,cholesterol,low density lipoprotein and insulin in the model cells were determined by enzyme method;（5）the gene expression of PI3K,Akt and IRS-1 in the model cells was determined by Q-PCR;（6）the protein expression of PI3K,Akt and IRS-1 in the model cells was determined by Western blot.Results:（1）compared with Hep G2 cells in blank group,TC,TG and LDL-C in ir-hepg2 model group were significantly increased（P<0.01）,and the m RNA and protein expressions of PI3K,Akt and IRS-1 were significantly decreased（P<0.01）.The difference was significant,and the model was successfully established;（2）compared with model group,TC and TG in high,medium and low dose groups of Dahuang lianxiexin Decoction were lower,and the results were statistically significant（P<0.01）,there was no significant difference in LDL-C of low-dose group（P>0.05）,but there was no significant difference between high-dose group and middle dose group（P>0.05）)The m RNA and protein expressions of PI3K,Akt and IRS-1 in high,medium and low dose groups of Dahuang Huanglian Xiexin Decoction were increased（P<0.05）,the results were statistically significant;（3）compared with Jinqi hypoglycemic group,TG and LDL-C in high,medium and low dose groups of Dahuang Huanglian Xiexin Decoction were significantly decreased（P<0.05）,the difference was statistically significant,TC high and low dose groups were significantly decreased（P<0.05）,the difference was statistically significant The m RNA expression of PI3K and IRS-1 in high,medium and low dose groups of Dahuang Huanglian Xiexin Decoction were significantly increased（P<0.05）,the difference was significant;the protein expression of low PI3K,Akt and IRS-1 in high and low dose groups of Dahuang Huanglian Xiexin Decoction were significantly increased（P<0.05）,the difference was significant.Conclusion:（1）Dahuang Huanglian Xiexin Decoction can reduce the cholesterol（TC）,triglyceride（TG）and low density lipoprotein（LDL-C）of IR-Hep G2 cells,so as to improve the lipid metabolism of liver;（2）Dahuang Huanglian Xiexin Decoction can increase the m RNA and protein expression of PI3K,Akt and IRS-1 in IR-Hep G2 cells,thus regulating PI3K/Akt signaling pathway and reducing insulin resistance.（3）The experimental results of rhubarb Huanglian Xiexin Decoction medium dose group were significantly better than that of Jinqi hypoglycemic group,which made up for the blank of in vitro study on the signal pathway of rhubarb Huanglian Xiexin Decoction,and provided a new idea and method for the treatment of type 2 diabetes with traditional Chinese medicine. |
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