不同浓度葡萄糖对3T3-L_1细胞分化及insig-1和insig-2 mRNA表达的影响

Effect of different glucose concentrations on the expressions of insig-1 and insig-2 mRNA during the differentiation of 3T3-L_1 cells

目的:观察不同浓度葡萄糖对小鼠前脂肪细胞(3T3-L1细胞)分化及insig-1和insig-2 mRNA基因表达的影响,探讨insig基因在脂肪细胞分化与代谢中的作用。方法:将3T3-L1细胞分别在高葡萄糖浓度(25mol/LG.S)、低葡萄糖浓度(5.5mol/LG.S)和甘露醇(19.5mol/L甘露醇+5.5mol/LG.S)中诱导分化,利用油红"O"染色检测脂肪细胞分化,RT-PCR和原位杂交法检测insig-1和insig-2 mRNA、脂肪细胞脂肪酸结合蛋白(AP2)mRNA表达的动态变化。结果:随着3T3-L1细胞的分化,insig-1和insig-2 mRNA和AP2 mRNA表达逐渐上调,低糖诱导组及甘露醇对照组细胞分化程度显著低于高糖诱导组,但两组insig-1和insig-2 mRNA的表达较高糖诱导组明显上调(P<0.05),而AP2 mRNA表达则下调;低糖诱导组与甘露醇对照组细胞分化程度及各基因表达无明显差别。结论:葡萄糖浓度可影响脂肪细胞分化;低糖时脂肪细胞分化及脂质沉积相对受抑制,可能与insig基因表达上调有关。

Objective To determine the effect of different concentrations of glucose on the differentiation of 3T3-L1 and the expression of insig-1 and insig-2 mRNA, and to explore the effect of insulin-induced gene in the differentiation and formation of adipocytes and lipogenesis. Methods The 3T3-L1 cells were induced to differentiate in high glucose concentration （25 mol/L G. S） , low glucose concentration （5. 5 mol/L G. S）, and mannitol （ 19. 5 mol/L Mannitol ＋ 5. 5 mol/L G. S） , respectively. The differentiation of 3T3-L1 cells was examined by oil red ＂O＂ straining, and the expression of insig-1 , insig-2 mRNA and AP2 mRNA was examined by RT-PCR and in situ hybridization. Results With the differentiation of 3T3-L1 cells, the expression of insig-1 and insig-2 mRNA was gradually up-regulated. The expression of insig-1 and insig-2 mRNA significantly increased while AP2 mRNA decreased in the low glucose concentration inducing group and mannitol inducing group. In the high glucose concentration inducing group, the cell differentiation was poor P〈 0.05 ）. There was no difference in the differentiation of 3T3-L1 cells, Conclusion Different concentrations glucose concentration promotes insig-1 of the differentiation and lipogenesis of between the low glucose concentration and the mannitol group and in the expression of insig-1 and insig-2 and AP2 mRNA. of glucose may influence the cell differentiation and the low and insig-2 gene expression, which may lead to the inhibition preadipocytes.