HepG2 cell IR:Establishment of model and improvement by berberine / 中国药理学通报

ABSTRACT

Aim To establish insulin resistance cell model on HepG2 cells (human embryonic liver tumor cells )and investigate the effect of berberine hydro-chloride on insulin-resistant HepG2 (IR-HepG2 ) cells.Methods ① IR model was induced by respec-tively using 10 -9 ,10 -8 ,10 -7 ,10 -6 ,10 -5 ,10 -4 mol ·L-1 insulin with 25 mmol · L-1 glucose in HepG2 cells.② HepG2 cells were incubated with 2-NBDG (fluorescent labeled glucose)in a series of concentra-tion50,100,200,400,600,800 μmol·L-1 and a series of incubation time20,40,60,80,100 min, to select the optimum concentration of insulin and the optimum incubation concentration and time of 2-NBDG in HepG2 cells.The success of the model was deter-mined by detecting the consumption of glucose in the cell supernatant and the uptake of glucose in HepG2 cells.③To study the effect of berberine hydrochloride on improving insulin resistance on the cell level,met-formin and berberine hydrochloride were used in the IR cells.Results Six concentrations of insulin induced the IR model in different degrees.Although 10 -4, 10 -5 mol·L-1 insulin was significant,a large amount of cells died.10 -6 mol·L-1 insulin was effective and had high survival rate of HepG2 cells,which had sta-tistical significance compared with the normal group. When the incubation concentration of 2-NBDG was more than 100 mol·L-1 ,the fluorescence intensity of the cells was significantly different from the normal group.When the incubation time of 2-NBDG was more than 20 min,fluorescence intensity was significantly different from the normal group.When the incubation time of 2-NBDG was more than 100 min,the fluores-cence quenching phenomenon was obvious in the cells. Berberine hydrochloride and metformin significantly in-creased the glucose consumption and glucose uptake in cell supernatant, which had statistical significance compared with the model group.Conclusions Using 10 -6 mol · L-1 insulin induced IR model in HepG2 cells,the optimum incubation concentration and incu-bation time of 2-NBDG is 200 μmol·L-1 and 80 min, respectively.Berberine hydrochloride and metformin have obvious effect on improving IR in HepG2 cells.