本文關(guān)鍵詞： N-乙酰半胱氨酸 亞砷酸鈉 HC心肌細(xì)胞 細(xì)胞凋亡 細(xì)胞自噬　出處：《中國計(jì)劃生育學(xué)雜志》2015年05期 　論文類型：期刊論文

【摘要】：目的:探討N-乙酰半胱氨酸(NAC)抑制亞砷酸鈉誘導(dǎo)H9C2細(xì)胞凋亡和自噬作用及機(jī)制。方法:用亞砷酸鈉誘導(dǎo)H9C2細(xì)胞構(gòu)建凋亡和自噬模型。采用CCK-8比色法檢測細(xì)胞的存活率,Hoechst 33258核染色法觀察凋亡細(xì)胞的形態(tài)和數(shù)量改變.雙氯熒光素染色法檢測細(xì)胞內(nèi)活性氧(ROS)水平,Western Blot法測定蛋白P53、Bcl-2、Bax及LC3的表達(dá)水平,以及磷酸化ERK1/2蛋白及ERK1/2蛋白總量的表達(dá)水平。結(jié)果:與對照組相比,15μM亞砷酸鈉處理的H9C2細(xì)胞存活率顯著降低.細(xì)胞內(nèi)ROS水平明顯增加.凋亡細(xì)胞數(shù)量明顯增多,自噬標(biāo)志蛋白LC3-Ⅱ與LC3-Ⅰ比值明顯增大(P0.01)。1 mM NAC預(yù)處理后顯著減少H9C2細(xì)胞內(nèi)ROS的生成和促凋亡蛋白P53和Bax的表達(dá),誘導(dǎo)抗凋亡蛋白Bcl-2表達(dá)升高,同時降低自噬相關(guān)蛋白LC3-Ⅱ與LC3-Ⅰ的比值。亞砷酸鈉處理誘導(dǎo)ERK1/2蛋白磷酸化(P0.01),而NAC和亞砷酸鈉共處理抑制了ERK1/2蛋白的磷酸化。結(jié)論:亞砷酸鈉降低H9C2細(xì)胞的存活率,增加細(xì)胞內(nèi)ROS的生成,誘導(dǎo)細(xì)胞凋亡和自噬。NAC通過降低H9C2細(xì)胞內(nèi)的ROS水平和抑制ERK1/2蛋白的磷酸化拮抗亞砷酸鈉誘導(dǎo)的細(xì)胞凋亡和自噬。
[Abstract]:Objective: to study the N-acetylcysteine (NAC). Methods: the model of apoptosis and autophagy was established by sodium arsenite induced H9C2 cells. The survival rate of H9C2 cells was determined by CCK-8 colorimetric assay. The morphologic and quantitative changes of apoptotic cells were observed by Hoechst 33258 nuclear staining and the intracellular reactive oxygen species (Ros) levels were detected by dichlorofluorescein staining. Western Blot assay was used to detect the expression of protein P53, Bcl-2, Bax and LC3. Results: compared with the control group, the expression levels of phosphorylated ERK1/2 protein and total ERK1/2 protein were measured. The survival rate of H9C2 cells treated with 15 渭 M arsenite was significantly decreased, the intracellular ROS level was significantly increased, and the number of apoptotic cells was significantly increased. The ratio of autophagy marker protein LC3- 鈪，

本文編號：1463665