人源肝癌HepG2细胞胰岛素抵抗和逆转模型中FoxO1 mRNA和蛋白表达水平及其意义

发布时间：2018-05-24 04:35

本文选题：人源肝癌HepG细胞 + 胰岛素抵抗　；参考：《吉林大学学报(医学版)》2017年04期

【摘要】：目的:探讨人源肝癌HepG2细胞胰岛素抵抗(IR)模型(HepG2/IR)和逆转模型(HepG2/IRPH)中叉头状转录因子O1(FoxO1)的mRNA和蛋白表达水平,阐明该基因参与IR的作用机制。方法:选择人源肝癌HepG2细胞,采用终浓度1×10~(-10)、1×10~(-9)、1×10~(-8)、1×10~(-7)、1×10~(-6)和1×10~(-5) mol·L~(-1)胰岛素诱导24、48和72h,对照组细胞不加胰岛素,用葡萄糖氧化酶法检测上清液中葡萄糖水平,计算各组细胞的葡萄糖消耗量,以确定IR模型建立的最佳诱导条件;采用终浓度为0.156、0.313、0.625、1.250、2.500、5.000、10.000、20.000mmol·L~(-1)盐酸吡咯列酮(PH)诱导HepG2建立逆转抵抗模型,同时设立对照组,MTT法检测细胞增殖活性,确定PH的最佳逆转浓度;利用Real-time PCR法和Western blotting法检测各组细胞FoxO1mRNA和蛋白表达水平。结果:1×10-7 mol·L~(-1)胰岛素作用36h,葡萄糖消耗量减少45.84%,发生明显的IR,与对照组比较差异有统计学意义(P0.05)。与对照组比较,1.25mmol·L~(-1)PH逆转抵抗24h,细胞增殖活性无明显变化(P0.05)。人源肝癌HepG2细胞发生IR时,与对照组比较,FoxO1mRNA和蛋白的表达水平均明显升高(P0.01);IR模型逆转后,与对照组比较,FoxO1mRNA和蛋白表达水平差异无统计学意义(P0.05)。结论:人源肝癌HepG2细胞IR与FoxO1mRNA的表达有关联性,FoxO1mRNA表达水平检测有可能作为胰岛素增敏剂的药效评估指标,降低FoxO1mRNA表达水平可作为2型糖尿病治疗的新靶点。
[Abstract]:Objective: to investigate the expression of mRNA and protein in HepG2 cell line of human hepatocellular carcinoma (HepG2) model of insulin resistance (IRR) and reverse model HepG2 / IRPH), and to elucidate the mechanism of this gene involved in IR. Methods: human hepatocellular carcinoma (HepG2) cells were induced by insulin for 2448 and 72 hours with 1 脳 10 ~ (-10) -10 ~ (-10) -10 ~ (-10) -10 ~ (-9) mol) and 1 脳 10 ~ (10 ~ (-7) mol / L ~ (-6) and 1 脳 10 ~ (10 ~ (-5) mol / L ~ (-1) for 24 48 h and 72 h, respectively. Glucose level in the supernatant was measured by glucose oxidase method, and the glucose consumption of the cells in each group was calculated. In order to determine the optimal induction conditions of IR model, the reverse resistance model was established by HepG2 induced by final concentration of 0.156 (0.3130.625) 1.250 (2.500,5.000,10.000mmol / L) pyrrolidone hydrochloride (PHH), and the control group was set up to detect the cell proliferation activity and determine the optimal reverse concentration of PH. The expression of FoxO1mRNA and protein was detected by Real-time PCR and Western blotting. Results when treated with 1 脳 10 ~ (-7) mol / L ~ (-1) insulin for 36 h, glucose consumption decreased by 45.84%, and there was a significant difference between control group and control group (P 0.05). Compared with the control group, 1.25mmol L~(-1)PH reversed the resistance for 24 h, but the cell proliferation activity did not change significantly (P 0.05). The expression level of FoxO1 mRNA and protein in HepG2 cells of human hepatocellular carcinoma was significantly higher than that of control group, and the expression level of FoxO1 mRNA and protein was not significantly different from that of control group. Conclusion: there is a correlation between IR and FoxO1mRNA expression in HepG2 cells of human hepatocellular carcinoma. The detection of FoxO1 mRNA expression may be used as an index to evaluate the pharmacodynamics of insulin sensitizer, and the decrease of FoxO1mRNA expression level may be a new target for the treatment of type 2 diabetes mellitus.
【作者单位】：兰州大学第二医院内分泌代谢科;兰州大学基础医学院医学实验中心;
【基金】：中央高校基本科研业务费专项资金资助课题(861174) 甘肃省科技厅自然科学基金青年科研基金资助课题(1308RJYA065) 甘肃省卫计委卫生行业计划项目资助课题(GSWSKY-2015-64)
【分类号】：R587.1

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