EGCG上调miR-29抑制APP_(695)突变的SH-SY5Y神经细胞Aβ生成研究

EGCG Attenuates Amyloid-β Generation Via Upregulation of mi R-29 in SH-SY5Y Cells with Mutant APP_(695)

目的:探讨没食子儿茶素没食子酸酯(EGCG)对含有APP_(695)突变的SH-SY5Y神经细胞中淀粉样肽(amyloid-β,Aβ)生成的影响及其机制。方法:将SH-SY5Y细胞分为正常组(Control)、模型组(含有APP_(695)突变的细胞,APP)和给药组(APP+10、20、30μmol·L-1的EGCG,24 h);免疫荧光细胞化学法检测各组细胞中APP的表达;MTT法检测各组细胞的存活能力;ELISA法检测各组Aβ_(1-42)的浓度;RT-PCR检测各组mi R-29的表达;RT-PCR和ELISA法检测各组BACE1(β-分泌酶)m RNA水平与蛋白水平的表达。结果:模型组中APP高表达;与模型组比较EGCG明显改善细胞形态,促进神经细胞的存活能力,降低Aβ_(1-42)的浓度,上调mi R-29的表达以及在m RNA与蛋白水平上降低BACE1的表达。结论:EGCG能抑制APP_(695)突变所导致的细胞Aβ生成,其作用机制可能与上调mi R-29的表达及下调BACE1的表达有关。

Objective：To investigate the effect of EGCG on Amyloid-β generation in SH-SY5 Y cells with mutant APP695 and its mechanisms. Methods：The cells were randomly divided into three groups,control group,model group（cells with mutant APP695）,medicine group（APP plus 10,20,30 μmol·L-1EGCG for 24 h）. The expression of APP was confirmed by immunofluesence,cell viability was detected by MTT assay,concentration of Aβ1-42 was assessed by ELISA,mi R-29 expression was determined by RTPCR,and BACE1 m RNA level and protein level among all group were assessed by RT-PCR and ELISA. Results：APP expression was significantly higher in model group. Compared with model groups,EGCG can improve the cells morphology,promote cells viability,decrease the concentration of Aβ1-42,upregulate the expression of mi R-29 and downregulate the expression of BACE1 in m RNA level and protein level. Conclusions：EGCG can inhibit Aβ generation in SH-SY5 Y cells with mutant APP695,which may be attributed to upregulation of mi R-29 and downregulation of BACE1.