MG132对SH-SY5Y细胞Aβ水平的影响

发布时间：2018-11-22 11:57

【摘要】：目的:观察蛋白酶体抑制剂MG132诱导SH-SY5Y细胞凋亡及调节β-淀粉样蛋白(beta-amyloid protein,Aβ)生成的作用,并对其机制进行探讨。方法:培养SH-SY5Y细胞,MG132对细胞进行处理,浓度分别为2.5μmol/L、5μmol/L和10μmol/L,24 h后检测各项指标。MTT法检测细胞活力;流式细胞术检测细胞凋亡;ELISA法检测细胞Aβ_(1-40)和Aβ_(1-42)水平;Western blot检测淀粉样蛋白前体蛋白(amyloid precursor protein,APP)、β-分泌酶(BACE1)、早老蛋白1(presenilins 1,PS1)、早老蛋白2(presenilins 2,PS2)、nicastrin(NCT)和α-分泌酶(ADAM10)蛋白表达。结果:经MG132处理后,细胞活力明显下降,诱导细胞凋亡,随剂量增加凋亡率分别为36.97%、46.20%、50.50%;细胞Aβ_(1-42)和Aβ_(1-40)蛋白水平明显上升;APP及Aβ代谢相关蛋白PS1、PS2和BACE1表达均出现剂量依赖性的增加,而ADAM10表达呈剂量依赖性降低;NCT水平变化不明显。结论:MG132能诱导神经细胞凋亡,通过影响Aβ生成途径关键蛋白而促进Aβ的产生,说明蛋白酶体活性下降可通过调节Aβ途径参与阿尔茨海默病的发生。
[Abstract]:Aim: to investigate the role of proteasome inhibitor MG132 in inducing apoptosis of SH-SY5Y cells and regulating the production of 尾 -amyloid protein (beta-amyloid protein,A 尾) and its mechanism. Methods: SH-SY5Y cells were cultured and treated with MG132 at concentrations of 2.5 渭 mol/L,5 渭 mol/L and 10 渭 mol/L,24 h, respectively. The cell viability was detected by MTT assay and apoptosis was detected by flow cytometry. The levels of A 尾 _ (1-40) and A 尾 _ (1-42) were detected by ELISA assay. The expressions of amyloid precursor protein (amyloid precursor protein,APP), 尾 -secretase (BACE1), presenilins _ 1 (presenilins _ 1), presenilins _ 2 (), nicastrin (NCT) and 伪 -secretase (ADAM10) were detected by Western blot. Results: after treated with MG132, the cell viability decreased and apoptosis was induced. The apoptotic rates were 36.97% and 46.20%, respectively, and the protein levels of A 尾 _ (1-42) and A 尾 _ (1-40) increased significantly. The expression of APP and A 尾 metabolism-related protein PS1,PS2 and BACE1 increased in a dose-dependent manner, while the expression of ADAM10 decreased in a dose-dependent manner, while the level of NCT did not change significantly. Conclusion: MG132 can induce neuronal apoptosis and promote the production of A 尾 by affecting the key protein of A 尾 pathway, which indicates that the decrease of proteasome activity may be involved in the pathogenesis of Alzheimer's disease by regulating A 尾 pathway.
【作者单位】：泰山医学院药学院;山东省高校动脉粥样硬化重点实验室泰山医学院动脉粥样硬化研究所;
【基金】：国家自然科学基金资助项目(No.81441111;No.81302202) 山东省自然科学基金资助项目(No.ZR2011HM044) 山东省卫生厅资助项目(No.2013WS0313) 泰安市科技计划资助项目(No.2015NS2072)
【分类号】：R749.16

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