摘要
在体外建立胰岛素抵抗肝细胞模型,探讨在胰岛素抵抗状态下成纤维细胞生长因子(FGF)-21对模型细胞糖代谢的影响及机制.将HepG2细胞置于10-7mol/L的胰岛素培养基中培养24h,建立胰岛素抵抗细胞模型.分别用不同浓度的胰岛素和FGF-21处理模型细胞,采用葡萄糖氧化酶-过氧化物酶(GOD-POD)法检测细胞对葡萄糖的摄取情况,并检查胰岛素与FGF-21的协同作用.利用实时荧光定量PCR检测FGF-21对模型细胞葡萄糖转运蛋白1(GLUT1)mRNA表达的影响,蒽酮法检测模型细胞糖原合成量,探讨FGF-21对胰岛素抵抗细胞模型葡萄糖摄取的影响及机制.结果发现,用高浓度胰岛素处理HepG2细胞24h后,细胞对胰岛素的敏感性显著降低,说明成功建立了胰岛素抵抗细胞模型,抵抗状态可维持48h,未发现细胞形态学变化.FGF-21能改善胰岛素抵抗模型细胞的葡萄糖摄取,参与肝糖原的合成,并与胰岛素产生协同作用.实时荧光定量PCR结果发现,FGF-21作用模型细胞后,细胞的GLUT1mRNA表达量显著增加,说明FGF-21促进模型细胞摄取葡萄糖的作用机制与其增加GLUT1的表达有关.
Insulin resistance in insulin sensitive organ results in metabolic disorder such as hyperglycemia, hyperinsulinemia and hyper triglyceridemia which are common features of type 2 diabetes. Insulin resistance in liver cells mainly causes impaired glycogen synthesis, failed to suppress glucose production which is the major contribution to hyperglycemia. FGF-21 as a new metabolic regulator can control fasting blood glucose. The mechanism of FGF-21 effects on regulating plasma glucose has little to known. In order to establish an in vitro insulin resistant model of liver cells and evaluate the effects and mechanism of FGF-21 on glucose metabolism in the cell model, HepG2 cells were incubated with 10^-7 mol/L insulin for 24 h to build insulin-resistant cell model. To evaluate the cells for insulin resistance, the cells were stimulated with fresh insulin for 24 h and the glucose uptake by these cells was carried out. The insulin-resistant cells were treated with different concentrations of FGF-21 for 24 h and insulin-treated cells were used as a control. The glucose uptake by the cells was detected by the method of glucose oxidizes/peroxides （GOD-POD）; the synergy between insulin and FGF-21 was evaluated. The mRNA expression of GLUT1 in the insulin-resistant cells was detected by the real-time PCR. Glycogen synthesis of the cells was examined by the anthrone method. The results showed that HepG2 cells treated with 10^-7 mol/L insulin for 24 h became resistant to insulin and the insulin resistance status was maintained for 48 h without change of cell morphology. FGF-21 could stimulate glucose consumption of the insulin-resistant model in a dose-dependent manner. The glucose consumption and glycogen synthesis of the insulin-resistant model were significantly improved by FGF-21 treatment. FGF-21 showed strong synergy with insulin in glucose uptake and glycogen synthesis of the model cells. While the cells became resistant to insulin, FGF-21 could increase the mRNA expression of GLUT1. Thus, It is concluded that FGF-21
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2009年第10期1327-1333,共7页
Progress In Biochemistry and Biophysics
基金
黑龙江省科技厅重点攻关项目(2006G0461-00)~~
