The Effect Of Kun Dan Formula On Glycolipid Metabolism Of Metabolic Syndrome

ObjectiveThe aim of our study is to explore the effect and molecular mechanism of Kun Dan formula(KD)on metabolic syndromeby using insulin-resistant HepG2 cell model and a rat model of metabolic syndrome.Methods1.We used insulin to stimulate HepG2 cells to establish insulin-resistant HepG2 cell model and then divided into six groups:normal group,model group,metformin group,low-dosed KD group,middle-dosed KD group and high-dosed KD group.After drug administration for 24 h,MTT was used to test the survival rate of insulin-resistant HepG2 cells,and the glucose consumption of insulin-resistant HepG2 cells was determined according to the manufacturer’ s requirements.Further,real-time quantitative PCR was used to test the expression of AMPKα2、GLUT2,G6Pase and IRS-2mRNA,and western bloting was used to examine the protein expression of PI3K,AMPK a 2,p-AMPK(Thr172),Akt and p-Akt(Ser473)in HepG2 cells.2.We used high fat diet to establish a rat model of metabolic syndrome.The rats were divided into six groups:normal group,model group,metformin group,low-dosed KD group,middle-dosed KD group and high-dosed KD group.Six weeks later,the serum and liver tissue were collected for the analysis of body weight,Lee’ sindex and histological examination,the serum cholesterol,triglyceride,LDL-C,HDL-C,insulin and glucose level.Further,AMPK,p-AMPK(Thr172),PI3K,AKT and p-AKT(Ser473)protein expression in liver tissue were determined by western bloting.What’ s more,AMPK α 2,GLUT2 and G6Pase mRNA expression in liver tissue were also determined by qPCR.Results1.After administration with 1×10-9 mol·L-1 insulin for 36 h,the glucose consumption of HepG2 cells was lowed suggesting that insulin-resistant HepG2 cells were successfully established in our study.2.The survival rate of insulin-resistant HepG2 cells is higher than 97%and there is no。ignificant difference among all groups.The results from qPCR shown that KD could increase the expression of AMPKα2,GLUT2 and IRS-2 mRNA,and decrease the expression of G6Pase mRNAin HepG2 cells.Western bloting showed that KD could also increase the protein expression of PI3K,p-AMPK(Thr172)and p-Akt(Ser473)of HepG2 cells.3.KD could effectively inhibit the body weight and obesity of rat、with metabolic syndrome.Further,KD could attenuate the level of serum cholesterol,triglyceride and LDL-C and increase the serum HDL-C level of rats with metabolic syndrome.What‘s more,KD could enhance the insulin sensitivity and attenuate the histological changes in the liver tissue of rats with metabolic syndrome.Western bloting and qPCR analysis showed that KD could efficiently increase the protein expression of p-AKT(Thr172),p-AMPK(Ser473)and PI3K and the mRNA expression of AMPK and GLUT2 in the liver tissue of rats with high-fat diet-induced metabolic syndrome.Further,KD could inhibit the mRNA expression of G6Pase genes.Conclusions1.1× 10-9 mol·L-1 insulin can induce insulin resistance in HepG2 cells.2.The KD is optimized,the compatibility of medicines is reasonable,the effect of invigorating spleen and tonifying deficiency,eliminating phlegm and removing blood stasis,soothing liver and regulating Qi,are played together,and good treatment effect is achieved for the basic pathogenesis of deficiency,phlegm and blood stasis manifested by metabolic syndrome.3.KD can enhance the glucose consumption of insulin-resistant HepG2 cells which is related to the activation of AMPK and PI3K/AKT signaling pathways.4.KD can inhibit the body weight and obesity,improve the hyperglycemia and hyperlipidemia,enhance insulin sensitivity and improve the hepatic steatosis of rats with metabolic syndrome.The molecular mechanism is related to the up-regulated expression of AMPK and PI3K/AKT signaling pathways.