【摘要】：目的:觀察蛋白酶體抑制劑MG132誘導SH-SY5Y細胞凋亡及調節(jié)β-淀粉樣蛋白(beta-amyloid protein,Aβ)生成的作用,并對其機制進行探討。方法:培養(yǎng)SH-SY5Y細胞,MG132對細胞進行處理,濃度分別為2.5μmol/L、5μmol/L和10μmol/L,24 h后檢測各項指標。MTT法檢測細胞活力;流式細胞術檢測細胞凋亡;ELISA法檢測細胞Aβ_(1-40)和Aβ_(1-42)水平;Western blot檢測淀粉樣蛋白前體蛋白(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)蛋白表達。結果:經(jīng)MG132處理后,細胞活力明顯下降,誘導細胞凋亡,隨劑量增加凋亡率分別為36.97%、46.20%、50.50%;細胞Aβ_(1-42)和Aβ_(1-40)蛋白水平明顯上升;APP及Aβ代謝相關蛋白PS1、PS2和BACE1表達均出現(xiàn)劑量依賴性的增加,而ADAM10表達呈劑量依賴性降低;NCT水平變化不明顯。結論:MG132能誘導神經(jīng)細胞凋亡,通過影響Aβ生成途徑關鍵蛋白而促進Aβ的產(chǎn)生,說明蛋白酶體活性下降可通過調節(jié)Aβ途徑參與阿爾茨海默病的發(fā)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 偽 -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.
【作者單位】： 泰山醫(yī)學院藥學院;山東省高校動脈粥樣硬化重點實驗室泰山醫(yī)學院動脈粥樣硬化研究所;
【基金】：國家自然科學基金資助項目(No.81441111;No.81302202) 山東省自然科學基金資助項目(No.ZR2011HM044) 山東省衛(wèi)生廳資助項目(No.2013WS0313) 泰安市科技計劃資助項目(No.2015NS2072)
【分類號】：R749.16

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