Effects Of Berberine On The Expression Of Hepatocyte Nuclear Factors MRNA And The Activity Of Glucose Metabolic Related Enzymes In HepG2 Cells

Objective:1. To observe the effect of berberine in different concentrations on the growth of human tumer cell strain HepG2, and find an optimal concentration of berberine for in vitro intervention.2. The expression of hepatocyte nuclear factors (HNF1α,HNF1β,HNF4α,HNF6) mRNA and the activities of glucose metabolism related enzymes (glucokinase(GK), L-pyruvate kinase(L-PK), phosphoenolpyruvate carboxykinase (PEPCK)) in HepG2 cells were observed to investigate the possible mechanism of berberine improving diabetes.Methods:1. HepG2 cells were cultured in vitro with a variety of concentrations of berberine(0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30, 100, 300μmol/L), and the growth of HepG2 cells was determined with MTT method.2. HepG2 cells were incubated with a variety of concentration of berberine (0,0.1,0.3,1,3,10,30μmol/L) and 1mmol/L metformin for 24h. The expression of HNFs mRNA were detected by RT-PCR and the activities of glucose metabolic related enzymes were determined with enzyme kinetics method. Results:1. Effect of berberine on the growth of HepG2 was dose dependent. At a concentation less than 1μmol/L, berberine showed little influence on the cells; at a concentation less than 10μmol/L, berberine affected cell number to a certain degree; and at a concentation less than 10μmol/L, berberine inhibited growth of HepG2 cells markedly.2. Compared with the control group, the expression of HNFs mRNA were decreased to some extent, the activities of glucokinase (GK) and L-pyruvate kinase (L-PK) increased and of phosphoenolpyruvate carboxykinase (PEPCK) decreased on berberine group in a dose-dependent manner.Conclusion:1. Berberine at the concentration less than 10μmol/L has little affection on HepG2 cell proliferation, while it can inhibite the growth of HepG2 cells markedly when its concentration over 30μmol/L.2. Berberine decreased the expression of HNFs mRNA to some extent and regulated the activities of glucose metabolism related enzymes in a dose-depended manner in HepG2 cells, which might be one of the mechanisms of berberine improving diabetes.