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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。
AIM: To investigate the effect and mechanism of EGCG on the production of amyloid-β (Aβ) in SH-SY5Y neurons with APP_ (695) mutation. Methods: SH-SY5Y cells were divided into control group, model group (APP 695 mutation-containing cells, APP) and administration group (APP + 10, 20 and 30 μmol·L -1 EGCG for 24 h ). The expression of APP in each group was detected by immunofluorescence cytochemistry. The viability of cells in each group was detected by MTT assay. The concentration of Aβ 1-42 in each group was detected by ELISA. The expression of mi R-29 The expression of BACE1 (β-secretase) m RNA and protein in each group were detected by RT-PCR and ELISA. Results: Compared with model group, EGCG significantly improved cell morphology, promoted the survival of nerve cells, decreased the concentration of Aβ 1-42, up-regulated the expression of mi R-29 and increased the expression of m RNA and protein On the expression of BACE1. CONCLUSION: EGCG can inhibit Aβ production induced by APP 695 mutation. The mechanism may be related to up-regulation of mi R-29 expression and down-regulation of BACE1 expression.