摘要
目的观察体外合成成纤维细胞生长因子21(FGF21)对胰岛素抵抗(IR)肝细胞葡萄糖摄取的影响,并探讨其机制。方法体外合成具有一定稳定性的荧光标记的FGF21 mRNA。将肝细胞置于含34.4μg/mL重组胰岛素和1μg/mL地塞米松及10%FBS的RPMI-1640培养基中培养72 h,诱导建立IR肝细胞模型,将其分为模型对照组,胰岛素组和FGF21组。模型对照组不添加任何药物,胰岛素组加入1μg/mL重组胰岛素,FGF21组电转入FGF21 mRNA。采用葡萄糖氧化酶法测定3组细胞培养液中的葡萄糖含量,计算葡萄糖吸收量;实时荧光定量PCR方法检测3组葡萄糖转运蛋白1(GLUT1)mRNA表达,Western blotting法检测3组GLUT1蛋白表达。结果与模型对照组相比,FGF21组葡萄糖吸收量升高,细胞GLUT1 mRNA及蛋白表达升高(P均<0.05)。与模型对照组相比,胰岛素组葡萄糖吸收量不明显,细胞GLUT1 mRNA及蛋白表达无明显差异(P均>0.05)。结论体外合成的FGF21 mRNA可以改善IR肝细胞对葡萄糖的摄取,其机制与增加GLUT1表达有关。
Objective To observe the effect of fibroblast growth factors-21（ FGF21） on glucose uptake of insulin-resistant（ IR） liver cells. Methods The fluorescently labeled FGF21 mRNA with certain stability was synthesized in vitro.The liver cells were cultured for 72 h in RPMI-1640 medium supplemented with 34. 4 μmol/L recombinant insulin and 1 μmol/L dexamethasone to establish IR cell models,and then IR cell models were divided into the model control group,insulin group,and FGF21 group. We did not add any drugs into the model control group. The insulin group was treated with 1 μmol/L recombinant insulin and FGF21 group was transfected with FGF21 mRNA. The cell glucose uptake in the three groups was measured by glucose oxidase（ GOD-POD） assay. The mRNA and protein expression of glucose transporter 1（ GLUT1） was detected by real-time fluorescent quantitative PCR and Western blotting. Results Compared with the model control group,the glucose uptake of the FGF21 group increased and the expression of GLUT1 mRNA and protein increased（ all P〈 0. 05）. Compared with the model control group,the glucose uptake of the FGF21 group did not change significantly in the insulin group,and no significant difference was found in the GLUT1 mRNA and protein expression as compared with that of the model control group（ all P 〉0. 05）. Conclusion FGF21 mRNA synthesized in vitro can improve the glucose uptake of IR liver cells,and its mechanism is related to the increase of GLUT1 expression.
出处
《山东医药》
CAS
北大核心
2017年第26期5-8,共4页
Shandong Medical Journal
基金
天津市科技计划项目(14ZCZDCX00054)
