【摘要】：目的:觀察蛋白酶體抑制劑MG132誘導(dǎo)SH-SY5Y細(xì)胞凋亡及調(diào)節(jié)β-淀粉樣蛋白(beta-amyloid protein,Aβ)生成的作用,并對(duì)其機(jī)制進(jìn)行探討。方法:培養(yǎng)SH-SY5Y細(xì)胞,MG132對(duì)細(xì)胞進(jìn)行處理,濃度分別為2.5μmol/L、5μmol/L和10μmol/L,24 h后檢測(cè)各項(xiàng)指標(biāo)。MTT法檢測(cè)細(xì)胞活力;流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡;ELISA法檢測(cè)細(xì)胞Aβ_(1-40)和Aβ_(1-42)水平;Western blot檢測(cè)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(dá)。結(jié)果:經(jīng)MG132處理后,細(xì)胞活力明顯下降,誘導(dǎo)細(xì)胞凋亡,隨劑量增加凋亡率分別為36.97%、46.20%、50.50%;細(xì)胞Aβ_(1-42)和Aβ_(1-40)蛋白水平明顯上升;APP及Aβ代謝相關(guān)蛋白PS1、PS2和BACE1表達(dá)均出現(xiàn)劑量依賴性的增加,而ADAM10表達(dá)呈劑量依賴性降低;NCT水平變化不明顯。結(jié)論:MG132能誘導(dǎo)神經(jīng)細(xì)胞凋亡,通過(guò)影響Aβ生成途徑關(guān)鍵蛋白而促進(jìn)Aβ的產(chǎn)生,說(shuō)明蛋白酶體活性下降可通過(guò)調(diào)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(xué)院藥學(xué)院;山東省高校動(dòng)脈粥樣硬化重點(diǎn)實(shí)驗(yàn)室泰山醫(yī)學(xué)院動(dòng)脈粥樣硬化研究所;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(No.81441111;No.81302202) 山東省自然科學(xué)基金資助項(xiàng)目(No.ZR2011HM044) 山東省衛(wèi)生廳資助項(xiàng)目(No.2013WS0313) 泰安市科技計(jì)劃資助項(xiàng)目(No.2015NS2072)
【分類號(hào)】：R749.16

【相似文獻(xiàn)】

相關(guān)期刊論文 前3條

1 安麗榮,金連弘,趙育楨,傅松濱;人app基因轉(zhuǎn)染PC12細(xì)胞及效果檢測(cè)[J];解剖科學(xué)進(jìn)展;2005年02期

2 張晶晶;王法財(cái);王海萍;梁燕;沈玉先;;重組人tau蛋白對(duì)神經(jīng)細(xì)胞的毒性作用[J];安徽醫(yī)科大學(xué)學(xué)報(bào);2009年02期

3 ;[J];;年期

相關(guān)會(huì)議論文 前1條

1 鄔英全;于明;;Aβ_(25-35)致PC12細(xì)胞核酸、蛋白質(zhì)氧化損傷及MCI-186的保護(hù)作用[A];第九次全國(guó)神經(jīng)病學(xué)學(xué)術(shù)大會(huì)論文匯編[C];2006年

，

本文編號(hào)：2349311