绿茶提取物表没食子儿茶素没食子酸酯对酒精诱导SH-SY5Y细胞损伤的作用研究

摘要

Objective: To observe in vitro effects of green tea compound epigallocatechin-3-gallate (EGCG) on alcohol-induced SH-SY5Y cell injury and discuss the feasibility of alcohol-associated dementia (AAD) treatment with EGCG.Methods: SH-SY5Y neuron cells were treated with ethanol (100 mol/L) for 24 h in 37℃ for ethanol group. In EGCG + ethanol group, cells were treated with different doses of EGCG (1—100 µmol/L) and ethanol (100 mol/L) for 24 h in 37℃. For control group or EGCG group, cells were treated only with cell medium or same doses of EGCG. The cell viability was analyzed with MTT assay. The staining with Annexin-V APC/7- AAD was used to observe cell apoptosis and the cell morphology was observed by optical microscope.Results:hTe cell survival was improved by treatment of EGCG for 24 h in a dose-dependent manner (25—100 µmol/L). Compared with control group (4.90%±0.80%) or EGCG group (7.82%±0.68%), cell apoptosis was apparently induced by treatment of ethanol (100 mol/L) for 24 h (21.82%±1.27%,P<0.001). However, co-treated with EGCG (100 µmol/L), these effects of ethanol on SH-SY5Y cells was prevented (10.81%±0.31%,P<0.001). Conclusion:EGCG inhibits alcohol-induced SH-SY5Y cell apoptosis and improve cell survival, which suggests that EGCG may have potential values for AAD treatment.%目的：体外观察绿茶提取物表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate，EGCG)对酒精诱导SH-SY5Y细胞损伤的作用，初步探讨EGCG治疗酒精性痴呆(alcohol-associated dementia，AAD)的可行性。方法：以SH-SY5Y神经元细胞为实验对象，酒精组给予100 mol/L酒精37℃培养24 h， EGCG+酒精组在其培养液中同时加入不同浓度的EGCG(1~100µmol/L)及100 mol/L酒精，37℃培养24 h；另设相同浓度的EGCG对照组(EGCG组)及空白对照组(对照组)。采用四甲基偶氮唑蓝(MTT)比色法检测细胞存活率，以Annexin-V APC/7-AAD双染法检测细胞凋亡及光镜下观察细胞形态。结果：MTT结果证实EGCG(25~100µmol/L)作用24 h后可明显促进SH-SY5Y细胞存活，并呈浓度依赖关系(P<0.001)。流式检测及细胞形态观察结果证实酒精组细胞凋亡率(21.82%±1.27%)，明显高于对照组(4.90%±0.80%)(P<0.001)及EGCG组(7.82%±0.68%)(P<0.001)；EGCG(100µmol/L)+酒精组细胞凋亡率(10.81%±0.31%)，明显低于酒精组(P<0.001)。结论：EGCG可抑制酒精诱导SH-SY5Y细胞凋亡发生并促进细胞存活，提示EGCG对于AAD治疗具有潜在价值。