中国药理学通报2016，Vol.32Issue(12)：1750-1754,5.DOI:10.3969/j.issn.1001-1978.2016.12.022
HepG2细胞胰岛素抵抗模型建立及盐酸小檗碱改善胰岛素抵抗的实验研究
HepG2 cell IR:Establishment of model and improvement by berberine
摘要
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-tion:50,100,200,400,600,800 μmol·L-1 and a series of incubation time:20,40,60,80,100 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.关键词
盐酸小檗碱,HepG2细胞,胰岛素抵抗,细胞模型/葡萄糖摄取/实验研究Key words
berberine hydrochloride/HepG2 cells/in-sulin resistance/the cell model/glucose uptake/ex-perimentel research引用本文复制引用
龚菂,李芬,邹欣,王定坤,陆付耳,王开富..HepG2细胞胰岛素抵抗模型建立及盐酸小檗碱改善胰岛素抵抗的实验研究[J].中国药理学通报,2016,32(12):1750-1754,5.基金项目
国家自然科学基金资助项目 （）
