表没食子儿茶素没食子酸酯对稳定表达淀粉样前体蛋白的SH-SY5Y细胞抗炎抗凋亡作用研究

目的 探讨表没食子儿茶素没食子酸酯(epigallocatechingallate,EGCG)对稳定表达淀粉样前体蛋白(amyloid precursor protein, APP)的SH-SY5Y细胞抗炎抗凋亡的保护作用.方法 将SH-SY5Y细胞分为对照组、模型组(稳定表达APP的细胞)和EGCG组(APP+10、20、30 μmol/L的EGCG).免疫荧光细胞化学法观察APP在各组细胞中表达;MTT法检测细胞存活能力;TUNEL法检测细胞凋亡情况;RT-PCR法检测Bax、Bcl-2和Caspase-3表达;ELISA法测定IL-6和TNF-α浓度.结果 模型组中,APP高表达;与模型组比较,给予20 μmol/L EGCG的EGCG组明显提高SH-SY5Y细胞的存活率,减少细胞凋亡数目,下调Bax/Bcl-2比例及Caspase-3 mRNA表达,减少IL-6和TNF-α浓度.结论 EGCG对稳定表达APP的SH-SY5Y细胞有一定保护作用,可能是通过抗炎以及减少细胞凋亡发挥治疗作用.

A Study on the Anti-inflammatory and Anti-apoptosis Effect of EGCG on APP-expressing SH-SY5Y Cells

Objective To investigate the anti-inflammatory and anti-apoptosis effect of EGCG on human SH-SY5Y cells expressing amyloid precursor protein (APP) stably. Methods The SH-SY5Y cells were randomly divided into three groups including the control group, model group (SH-SY5Y cells expressing APP stably ), and EGCG group ( APP-expressing cells treated with 10, 20 and 30 μmol/L EGCG respectively). The expression of APP was confirmed by the immunofluoresence staining; the cell viability was assessed by MTT; the number of cell apoptosis was assessed by TUNEL; the mRNA expression of Bax, Bcl-2 and Caspase-3 were determined by RT-PCR; the concentration of IL-6 and TNF-α was assessed by ELISA. Results The APP expression was higher in the model group. Compared with the model group, The administration of 20 μmol/L EGCG promoted the survival rate of SH-SY5Y cells, reduced the number of apoptotic cells, downregulated the radio of Bax and Bcl-2 and the mRNA expression of Caspase-3, and decreased the concentration of IL-6 and TNF-α. Conclusion EGCG has protective effect on the APP-expressing SH-SY5Y cells and the therapeutic effect may be achieved by anti-inflammation and anti-apoptosis.