N-乙酰半胱氨酸抑制亚砷酸钠诱导心肌细胞凋亡和自噬作用的研究

发布时间：2018-01-25 20:49

  本文关键词： N-乙酰半胱氨酸 亚砷酸钠 HC心肌细胞 细胞凋亡 细胞自噬　出处：《中国计划生育学杂志》2015年05期 　论文类型：期刊论文

【摘要】：目的:探讨N-乙酰半胱氨酸(NAC)抑制亚砷酸钠诱导H9C2细胞凋亡和自噬作用及机制。方法:用亚砷酸钠诱导H9C2细胞构建凋亡和自噬模型。采用CCK-8比色法检测细胞的存活率,Hoechst 33258核染色法观察凋亡细胞的形态和数量改变.双氯荧光素染色法检测细胞内活性氧(ROS)水平,Western Blot法测定蛋白P53、Bcl-2、Bax及LC3的表达水平,以及磷酸化ERK1/2蛋白及ERK1/2蛋白总量的表达水平。结果:与对照组相比,15μM亚砷酸钠处理的H9C2细胞存活率显著降低.细胞内ROS水平明显增加.凋亡细胞数量明显增多,自噬标志蛋白LC3-Ⅱ与LC3-Ⅰ比值明显增大(P0.01)。1 mM NAC预处理后显著减少H9C2细胞内ROS的生成和促凋亡蛋白P53和Bax的表达,诱导抗凋亡蛋白Bcl-2表达升高,同时降低自噬相关蛋白LC3-Ⅱ与LC3-Ⅰ的比值。亚砷酸钠处理诱导ERK1/2蛋白磷酸化(P0.01),而NAC和亚砷酸钠共处理抑制了ERK1/2蛋白的磷酸化。结论:亚砷酸钠降低H9C2细胞的存活率,增加细胞内ROS的生成,诱导细胞凋亡和自噬。NAC通过降低H9C2细胞内的ROS水平和抑制ERK1/2蛋白的磷酸化拮抗亚砷酸钠诱导的细胞凋亡和自噬。
[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，

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