格列喹酮可促进成肌细胞葡萄糖摄取

目的 观察格列喹酮对小鼠成肌细胞(C2C12)葡萄糖摄取及葡萄糖转运相关蛋白葡萄糖转运体4表达的影响.方法 采用格列喹酮干预体外培养的C2C12细胞,将细胞分为空白组、胰岛素组(1×10-7mol/L胰岛素)、格列喹酮组(1×10-5mol/L格列喹酮)、格列喹酮+胰岛素组(1×10-5mol/L格列喹酮+1×10-7mol/L胰岛素).应用葡萄糖氧化酶法检测细胞培养基中葡萄糖水平,DMEM组葡萄糖含量平均值减去其余各孔葡萄糖含量算得各孔细胞葡萄糖消耗量;用MTT法检测细胞活力;利用液闪计数法检测C2C12细胞葡萄糖摄取量;采用半定量实时聚合酶链反应(RTPCR)检测细胞中葡萄糖转运体4 mRNA水平.应用单因素方差分析进行数据统计.结果 与空白组相比,格列喹酮组C2C12细胞葡萄糖消耗量明显增加[(11.0±0.5)vs(7.9±0.6)mmol/L,q=0.36,P＜0.01];格列喹酮+胰岛素组C2C12细胞葡萄糖消耗量较胰岛素组增多[(13.8±0.7)vs(12.3±0.6)mmol/L,q=0.72,P＜0.01].胰岛素组、格列喹酮组C2C12细胞对葡萄糖的摄取均增加,分别为空白组的(1.68±0.09)、(1.52±0.23)、(1.23±0.16)倍(q值分别为14.78、11.30、5.00,均P＜0.01);格列喹酮组C2C12细胞葡萄糖摄取增加较格列苯脲组明显(q=6.30,P＜0.01),格列喹酮+胰岛素组C2C12细胞葡萄糖摄取为空白组的(1.93±0.12)倍(q=13.04,P＜0.01),且明显高于胰岛素组(q=5.43,P＜0.01).格列喹酮组C2C12细胞葡萄糖转运体4 mRNA水平与空白组比较无明显差异.结论 格列喹酮可促进C2C12细胞对葡萄糖的摄取,增加胰岛素刺激的葡萄糖摄取,并未增加C2C12细胞葡萄糖转运体4基因表达.

Effects of gliquidone on glucose uptake in C2C12 cells

Objective To investigate the effects of gliquidone on glucose uptake and the expression of glucose transporter 4 (GLUT4) in C2C12 myotubes. Methods Cultured C2C12 myotubes were used for glucose consumption studies. The groups were as follows:control group, 1 × 10 -5 mol/L gliquidone group, 1 × 10 -7 mol/L insulin group and 1 × 10 -5 mol/L gliquidone + 1 × 10-7 mol/L insulin group. Glucose concentrations in medium were determined by the glucose oxidase method. The amount of glucose consumption was calculated by the glucose concentrations of blank wells subtracting the remaining glucose in cell plated wells. Scintillation was used to detect the glucose uptake of C2C12 myotubes. The expression of GLUT4 mRNA was tested by real-time PCR (RT-PCR). One-way ANOVA was used for data analysis.Results Glucose consumption of C2C12 myotubes was significantly increased with 1 × 10-5 mol/L gliquidone ((11.0 ±0.5) vs (7.9 ±0.6) mmol/L, q =0.36, P ＜0.01). With the existence of gliquidone, the glucose-lowering effect of 1 × 10 -7 mol/L insulin was strengthened. Compared to the control cells, insulin, gliquidone, gliquidone + insulin, and glibenclamide increased 2-deoxyglucose uptake (q values were 14. 78,11.30 and 5.00, all P ＜0. 01) in C2C12 myotubes after incubation for 12 h with the indicated drug concentrations. The gliquidone-stimulated glucose uptake of mytubes was higher than that with the glibenclamide-stimulated (q = 6. 30,P ＜0. 01), and the insulin-stimulated glucose uptake was distinctly elevated in the presence of gliquidone (q =5.43 ,P ＜0. 01). Gliquidone did not enhance the expression of GLUT4 mRNA in C2C12 myotubes. Conclusion Gliquidone stimulates glucose uptake, increases the insulin-stimulated glucose uptake in C2C12 myotubes, but has no effect on the expression of GLUT4 mRNA.