姜黄素对swAPP HEK293细胞β样淀粉样蛋白生成的影响

Effect of curcumin on Aβ production in swAPP HEK293 cells

目的:在转染瑞典突变型淀粉样前体蛋白(APP)基因细胞(swAPP HEK293)中探讨姜黄素(Curcumin)对β样淀粉样蛋白(Aβ)生成的影响及初步机制。方法:以swAPP HEK293为细胞模型,首先以MTT的方法检测姜黄素在不同时间点(各个时间点)和浓度值(各个浓度值)时对细胞存活率的影响;选取无细胞毒性时的浓度值(5μmol/L)作用不同时间后,采用ELISA的方法检测其对Aβ的影响;选取抑制作用最强的浓度值和时间点进行后续试验,采用RT-PCR检测miR-153及APP mRNA的表达,采用Western blot检测APP蛋白的表达。结果:与对照组相比,姜黄素浓度≤5μmol/L组对细胞活性无毒性作用(P>0.05),姜黄素明显抑制Aβ的生成,差异有统计学意义(P<0.05),从而筛选姜黄素最佳作用浓度和时间为5μmol/L、24 h;姜黄素5μmol/L对APP mRNA表达水平无明显影响(P>0.05),而明显降低APP蛋白的表达(P<0.05);此外,姜黄素5μmol/L使细胞中miR-153的表达水平明显升高(P<0.05)。结论:在swAPP HEK293细胞中,姜黄素可能通过上调miR-153的表达及降低APP蛋白的表达进而抑制Aβ的生成。

Objective To investigate the effect of curcumin on Aβ production in swAPP HEK293 cells and its preliminary mechanism. Methods swAPP HEK293 was used as cell model, the effect of curcumin （at different time points and of concentration） on cell viability was accessed by MTT assay. After the cells were treated with non-eytotoxic concentration of 5 p, mol/L for different time, ELISA was used to detect Aβ production. The concentration and the time point of the strongest inhibitory effect were selected for the following tests. Real time PCR was employed to analyze miR-153 and APP mRNA expression, and Western blot was used to detect APP protein expression. Results As compared with the control group, curcumin of ≤ 5 μ mol/L had no toxicity effect on the cell viability （P 〉 0.05）. Curcumin significantly inhibited Aβ production （P 〈 0.05）. Therefore 5 μmol/L curcumin and 24 h were selected as the best concentration and time. Curcumin of 5 μmol/L had no obvious impact on APP mRNA expression （P 〈 0.05） , whereas markedly decreased APP protein expression. In addition, miR-153 level in the cells was significantly increased by 5 μ mol/L curcumin treatment （P 〈 0.05）. Conclusion Curcumin may inhibit Aβ production through up-regulating miR-153 level and reducing APP protein expression in swAPP HEK293 cells.