乙醇对HepG2细胞陷窝蛋白-1表达的影响

Effects of ethanol on the expression of caveolin-1 in HepG2 cells

目的探讨乙醇引起肝细胞膜的氧化应激以及此氧化应激机制是否与陷窝蛋白-1(CAV-1)相关。方法培养人肝癌HepG2细胞株,分别设实验组:不同浓度(100、200、500、1 000和1 500mmol/L)乙醇分别刺激不同时间(0~24h);正常对照组:用等量的1μmol/L无菌0.9%氯化钠注射液替代乙醇。四甲基偶氮唑盐比色法、乳酸脱氢酶(LDH)比色检测细胞生存率和细胞活性;选定200mmol/L浓度乙醇作为实验浓度,应用蛋白质印迹法、实时反转录聚合酶链反应(RT-PCR)检测CAV-1表达的动态变化。结果 200mmol/L浓度的乙醇未影响细胞的生存率。蛋白质印迹法检测200mmol/L乙醇在24h内致HepG2细胞CAV-1表达增多。结论乙醇刺激细胞能够诱导CAV-1表达增加。

Objective To investigate the ethanol-induced oxidative stress on liver cell membrane and whether the expression of caveolin -1 （CAV-1） can be influenced by the mechanism of oxidative stress. Methods Human HepG2 cell lines were cultured. Different molar concentration of ethanol （100,200,500,1 000 and 1 500 mmol/L） were applied at different times （0-24 h） in experimental group; Ethanol was substituted by the same amount of 1 μmol/L sterile saline in normal control group. The survival rate of the cells and cell viabilities were measured by 3-（4,5）-dimethylthiazo （-z-yl）-3,5-di-phenyltetrazoliumromide （MTT） and lactate dehydrogenase （LDH） colorimetric assays. The concentration of 200 mmol/L ethanol was selected as an experimental application of western blotting and study. Results The 200 mmol/L concentration of ethanol did not affect the survival rate of the cells. The western blotting suggested that, induced by the 200 mmol/L ethanol within 24 h, the expression of CAV-1 in HepG2 cells increased. Conclusion Moderate alcohol can stimulate liver cells, which induces the CAV-1 generation and accelerate the metabolism of ethanol.