叉头状转录因子O1与吡格列酮干预对肝癌HepG-2细胞葡萄糖消耗的影响  

Effects of forkhead transcription factor O1 and piglitazone on glucose consumption of HepG-2 cell line

作　　者：张鹏宇[1] 秦贵军[1] 王守俊[1] 张颖辉[1] 马笑堃[1] 樊大贝[1] 

机构地区：[1]郑州大学第一附属医院内分泌科,450052

出　　处：《中华糖尿病杂志》2011年第5期420-424,共5页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：河南省杰出青年项目（84100410016）

摘　　要：目的研究叉头状转录因子O1（FoxO1）及吡格列酮干预对高胰岛素诱导的肝癌HepG-2细胞葡萄糖消耗的影响及机制。方法1×10^-6mol／L胰岛素培养HepG．2细胞24h后，诱发培养基葡萄糖消耗减少建立细胞模型，将细胞分为对照组、空白质粒组、FoxOlsiRNA载体组、1×10^-5mol／L吡格列酮组。以普通培养基培养的细胞作为空白对照组。37℃、5％CO2培养箱中孵育24h，葡萄糖氧化酶法检测培养基中葡萄糖含量，逆转录-聚合酶链反应（RT—PCR）检测FoxO1mRNA和过氧化物酶体增殖物激活受体-γ（PPAR-γ）mRNA表达，Westernblot法检测PPAR-γ蛋白表达。将FoxO1mRNA、PPAR-γ mRNA、PPAR-γ蛋白表达与葡萄糖消耗量进行相关分析和曲线拟合。采用单因素方差分析及多样本均数两两比较进行统计学分析。结果高胰岛素培养24h较未诱导细胞培养基的葡萄糖消耗降低[分别为（1．36±0．03）和（2．93±0．05）mmol／L，P〈0．01]，细胞Fox01mRNA升高（分别为0．513±0．016和0．425±0．011，P〈0．05），PPAR-γmRNA降低（分别为0．260±0．025和0．441±0．012，P〈0．05），PPAR-γ蛋白降低（分别为0．312±0．032和0．600±0．046，P〈0．05）。在抑制FoxO1和吡格列酮干预后此趋势有所缓解，逐渐接近于空白对照组。FoxO1mRNA、PPAR-γ mRNA和PPAR-γ蛋白表达与葡萄糖消耗量高度相关。结论FoxO1表达升高或降低对葡萄糖代谢产生不利影响；增强PPAR-γ表达可有效恢复高胰岛素诱导的高糖，增加肝细胞的胰岛素敏感性。Objective To study the effect and mechanism of forkhead transcription factor （ FoxO1 ） and pioglitazone on the glucose utilization in HepG-2 cell line induced by high-level insulin. Methods HepG-2 ceils were induced by exposure to 1 ×10^-6 mol/L insulin for 24 hours. HepG-2 cells were divided into control group, blank plasmid group, FoxO1 small interfering RNA （siRNA） vector group, and pioglitazone medium group. The blank control group was cultured with normal medium. At 37℃, 5% CO2 for 24 hours, the glucose consumption was detected by glucose oxidase method, the expression levels of FoxOl mRNA and peroxisome proliferator-activated receptors-gamma （PPAR-7） mRNA were analyzed by RT-PCR, and the protein expression of PPAR-T was determined by Western blot. Single factor analysis of variance and SNK or Dunnet＇s test were used for statistic analysis. Results Compared with blank control group, the glucose consumption（ （ 1.36 ± 0. 03 ） vs （2.93 ± 0. 05 ） mmol/L, P 〈 0. 01 ） , FoxO1 mRNA （0. 260 ±0. 025 vs 0. 441 ±0. 012,P 〈0. 05） and protein expressions of PPAR-T decreased（0. 312 ±0. 032 vs 0. 600 -± 0. 046,P 〈 0. 05 ） , and the expression of FoxO1 mRNA increased （0. 513 ± 0. 016 vs 0. 425 ± 0. 011, P 〈0. 05） ; there was no difference between FoxO1 siRNA vector group, pioglitazone medium group and blank control group in these indices（ P 〉 0. 05 ）. After inhibiting FoxO1 by small interference RNA or induced by pioglitazone, these indices were closed to the indices of blank control group. The expressions of FoxO1 mRNA,PPAR-γ mRNA and PPAR-γ/ protein were highly correlated with the glucose consumption. Conclusions Inhibiting and increasing the expression of FoxO1 could produce adverse effects on glucose metabolism. Up-regulation PPAR-T expression could improve insulin sensitivity.

关 键 词：叉头转录因子类 过氧化物酶体增殖物激活受体 吡格列酮 HEPG-2细胞 

分 类 号：R735.7[医药卫生—肿瘤]