不同葡萄糖浓度对INS-1细胞PPARδ表达的影响

Effects of different concentrations of glucose on PPARδ expression in INS-1 cells

目的研究不同葡萄糖浓度对大鼠INS-1细胞PPARδ表达的影响,以初步阐明PPARδ在胰岛细胞糖脂代谢关联中的作用。方法分别用含3、11、20mmol/L葡萄糖的RPMI1640培养基,培养INS-1细胞24h后,冻融法裂解细胞,检测INS-1细胞内的甘油三酯水平。提取RNA和核蛋白,RT-PCR半定量检测PPARδmRNA,Westernblot检测PPARδ蛋白表达。结果随着葡萄糖浓度的升高INS-1细胞内甘油三酯含量增高,而PPARδmRNA和蛋白的表达均明显降低。结论葡萄糖促进,INS-1细胞内甘油三酯沉积,而PPARδ的表达下降可能是高糖条件下胰岛细胞脂质沉积的重要原因。

Objectives To observe the effects of glucose PPARδ in the rat insulinoma INS-1 cells so as to illustrate the tabolism with glucose metabolism in pancreatic cells. Methods at different concentrations on the expression of function of PPARδ in the cross-talk of lipid me-tabolism with glucose metabolism in pancreatic cells. Methods After cultured in RPMI1640 media containing 3, 11 and 20 mmol/L glucose respectively for 1 h, the cells were used to measure the content of intracellular triglyceride. The RNA and nuclear protein of INS-1 cells were extracted, and then the expression of PPARδ mRNA was detected by semi-quantity RT-PCR and the PPARδ protein was analyzed by Western blotting. Resuits With the increment of the concentrations of glucose, the content of intracellular triglyceride in the cultured cells was elevated, while the expressions of PPARδ mRNA and protein were both down-regulated. Conclusion Glucose promotes triglyceride deposition in INS-1 cells, and the down-regulation of PPARδ presumably contributes to the lipid deposition in INS-1 cells during hyperglycemic states.