附子多糖对胰岛素抵抗脂肪细胞模型GLUT4蛋白表达和转位的影响

The effect of Fuzi polysaccharides on the expression and translocation GLUT4 protein in insulin-resistance adipocytes

目的研究附子多糖对胰岛素抵抗脂肪细胞模型葡萄糖转运4(GLUT4)蛋白表达和转位的影响。方法将3T3-L1前脂肪细胞诱导分化为成熟的脂肪细胞,用高糖和高胰岛素联合诱导培养脂肪细胞,造成胰岛素抵抗脂肪细胞模型,实验设对照(CON)组、模型(MOD)组、附子多糖(FPS)组、罗格列酮(ROS)组,干预培养24 h,收集细胞,提取全细胞蛋白及细胞外膜蛋白,SDS-聚丙稀酰胺凝胶电泳后,通过Western blot方法检测各组全细胞蛋白及细胞外膜蛋白中GLUT4的含量。结果各组对GLUT4表达差异无统计学意义(P>0.05)。模型组转位显著降低,仅为正常脂肪细胞的30.4%,以模型组转位量为基值,各组分别与之相比计算GLUT4蛋白的相对定量,附子多糖组、罗格列酮组分别使GLUT4蛋白转位增加164%、301%,与模型组比较差异有统计学意义(P<0.01),附子多糖组显著低于罗格列酮组(P<0.01)。结论附子多糖对胰岛素抵抗脂肪细胞模型全细胞GLUT4蛋白表达无影响,但可促进其转位,其促进胰岛素抵抗脂肪细胞对葡萄糖的摄取可能与这一机理有关。

Objective To study the effect of Fuzi polysaccharides on the expression and translocation GLUT4 protein in insulin-resistance adipocytes. Methods 3T3-L1 proadipocytes were induced with drugs （Ins. ＋Dex. ＋IBMX.） and differentiated into adipocytes.High dose of glucose and insulin were employed to induce and culture adipocytes to establish insulin-resistance adipocytes model.The cells were divided into 4 groups：control group,model group,Fuzi polysaccharides group and rosiglitazone group.After 24 hours of FPS-treatment,the cells were harvested and the total exocellular and endocellular proteinsof the cells were extracted. SDS-PAGE and Western blot were used to identify the concentration of GLUT4 protein in all cell groups. Results There was no significant difference in the expression of GLUT4 protein among the four groups. But the translocation of GLUT4 was suppressed significantly in the model group, only 30.4% of the control group.Taking the level of GLUT4 translocation in model group as the base value （100%）.The translocation of GLUT4 increased 164% and 301% in FPS group and ROS group,respectively （compared with the model group,P〈0.01）.The translocation of GLUT4 in FPS group was significantly lower than that in ROS group （P〈0.01）. Conclusion Fuzi polysaccharides have no effect on the expression of GLUT4 protein in insulin-resistance adipocytes, but they can promote its translocation in the cells.