高浓度葡萄糖诱导HepG-2细胞的凋亡及其调控的相关机制

High concentrations of glucose-induced apoptosis in HepG-2 cells and their regulation mechanisms

目的探讨高浓度葡萄糖诱导HepG-2细胞的凋亡及其调控的可能机制。方法用含不同浓度葡萄糖的DMEM处理HepG-2细胞,分为正常组(糖浓度为5.5 mmol/L)和高浓度葡萄糖组(糖浓度分别为25、40、100、150、200mmol/L),采用MTT、Hoechst染色法观察细胞的增殖和凋亡状况;Western blot检测凋亡相关蛋白Bcl-2、Bak、Bax、p53、NF-κB的表达变化,并检测Caspase3的活性。结果高浓度葡萄糖能明显抑制细胞的体外增殖且呈剂量依赖性;Ho-echst染色显示高浓度葡萄糖能诱导细胞的凋亡,且随着葡萄糖浓度的升高凋亡现象越来越明显,Caspase3的活性呈上升趋势;Western blot显示高浓度葡萄糖能下调抑制凋亡作用的蛋白Bcl-2,上调促进凋亡的蛋白Bak、Bax、p53和NF-κB的表达(P<0.05)。结论高浓度葡萄糖能明显诱导HepG-2细胞的凋亡。

Objective To investigate high concentrations of glucose-induced apoptosis in HepG-2 cells and its possible regulation mechanism. Methods HepG-2 cells were treated with different concentrations of glucose, and were divided into normal group （ glucose concentration of 5.5 mmol/L） and high concentrations of glucose （ glucose concentration of 25, 40, 100, 150, 200mmol/L）. The cell proliferation and apoptosis of HepG-2 cell were analyzed by MTT, Hoechst staining;The expression of apoptosis-related protein Bcl-2, Bak, Bax, p53 and NF-kB were studied by Western blot, and the activity of Caspase3 was analyzed by Caspase3 kit. Results High concentrations of glucose could inhibit growth of HepG-2 cells ; Moreover high concentrations of glucose could also induce apoptosis with the concentration increased more obviously;Western blot showed that high concentrations of glucose could down-regulate Bcl-2, and increase Bak, Bax, p53 and NF-kB expression（P 〈 0. 05 ）. Conclusion High concentrations of glucose can induce apoptosis in HepG-2 cells.