高炳玉
GAO Bingyu

• 1:冀中能源井陉矿业集团有限公司总医院

摘要(Abstract)：

目的:探讨铅诱发心肌细胞氧化损伤及凋亡的分子机制。方法:无菌条件下取1~3日龄Wistar大鼠心肌细胞进行原代培养,选取不同浓度的醋酸铅(10、20、40、80、160及320μmol/L)处理心肌细胞48 h时,采用MTT法筛选细胞存活率为50%~80%的醋酸铅浓度;选择10、20、40、80及160μmol/L醋酸铅处理48 h的心肌细胞,采用氧化应激法测定心肌细胞的乳酸脱氢酶(LDH)、丙二醛(MDA)、活性氧(ROS)水平,采用Western blot测定心肌细胞凋亡相关蛋白Bcl-2、Bax、caspase-3及p53水平,仅加入培养基的心肌细胞作为空白对照组。结果:MTT结果显示,20~160μmol/L醋酸铅处理48 h的心肌细胞存活率为50%~80%;氧化应激结果显示,20~160μmol/L的醋酸铅处理的心肌细胞LDH、MDA、ROS水平高于空白对照组细胞(P<0.05);Western blot结果显示,与空白对照组比较,20~160μmol/L的醋酸铅处理的心肌细胞中Bcl-2表达量、Bcl-2与Bax的比值显著降低(P<0.05),Bax、caspase-3及p53表达量显著升高(P<0.05)。结论:20~160μmol/L的醋酸铅可以增加大鼠心肌细胞氧化应激水平,引起心肌细胞的氧化损伤,其机制可能与降低心肌细胞中抑制凋亡蛋白Bcl-2的表达量,提高心肌细胞中促凋亡基因Bax、caspase-3和p53的表达量有关。
Objective: To investigate the molecular mechanism of plumbum induced oxidative damage and apoptosis of myocardial cells. Methods: the myocardial cells were taken from Wistar rats of 1 ~3 d under aseptic conditions for primary culture. The different concentrations of lead acetate( 0,10,20,40,80,160,320 μmol/L) were selected for myocardial cells. The toxicity of lead acetate to myocardial cells was detected by MTT assay. Oxidative stress tests including the determination of lactate dehydrogenase( LDH),malondialdehyde( MDA) and reactive oxygen( ROS) levels in myocardial cells. Bcl-2,Bax,caspase-3 and p53 of myocardial apoptosis-related proteins were determined and compared by Western blot. Results: According to the results of MTT assay,20,40,80 and160 μmol/L lead acetate was selected as the toxic dose to ensure the survival rate of cells in 50 ~80%. Oxidative stress tests showed that the lead acetate( 20 ~ 80 μmol/L) increased the release amount of myocardial cell lactate dehydrogenase( LDH) in a dose-dependent manner and the content of myocardial cells malondialdehyde( MDA),and improved the production of myocardial cells reactive oxygen species( ROS),eventually leading to the oxidative damage of myocardial cells. Western blot results showed that the expression of anti-apoptotic protein Bcl-2 in the lead acetate( 20 ~ 80 μmol/L)treatment group decreased significantly,compared with the normal group( P < 0. 05). The expression of apoptotic protein Bax increased significantly( P < 0. 05),compared with the normal group. The ratio of Bcl-2 to Bax decreased significantly,promoting the apoptosis of myocardial cells. At the same time,the expression of apoptotic protein caspase-3 and p53 in myocardial cells was significantly higher than that in the normal group( P < 0. 05). Conclusion: Lead acetate( 20 ~ 160 μmol/L) can increase the production of LDH,MDA and ROS in myocardial cells,causing the oxidative damage of myocardial cells in rats. Furthermore,the expression of Bcl-2 in myocardial cells decreased,while the expression of apoptosis proteins Bax,caspase-3 and p53 in myocardial cells increased. At the same time,the ratio of bcl-2 to Bax decreased significantly,therefore lead acetate can promote the apoptosis of myocardial cells.

关键词(KeyWords)： 铅;心肌细胞;氧化应激;氧化损伤;细胞凋亡
lead;myocardial cells;oxidative stress;oxidative damage;apoptosis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31560270)

作者(Author): 高炳玉
GAO Bingyu

DOI: 10.19367/j.cnki.1000-2707.2018.08.005

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