学术论文

      PM2.5对血管内皮细胞氧化应激和凋亡的影响及机制

      Oxidative stress and apoptosis in endothelial cells exposed to PM2.5

      摘要:
      目的:从大气细颗粒物PM2.5对血管内皮细胞氧化应激和凋亡的影响,研究PM2.5对血管内皮细胞的毒性.方法:体外培养血管内皮细胞株EA.hy926,用不同浓度的PM2.5染毒24 h后,用CCK-8法测细胞的活性,用DCFH-DA荧光标记法检测细胞内氧自由基生成情况,用流式细胞术检测细胞凋亡率,然后用Western blot法检测凋亡相关蛋白细胞色素C、cleaved caspase-9和cleaved caspase-3的表达变化.结果:CCK-8法结果显示PM2.5对血管内皮细胞有明显的毒性,在浓度大于25 mg/L时可使EA.hy926细胞活性显著下降;PM2.5染毒24 h后可见DCFH-DA荧光染色增强,说明细胞内有大量的氧自由基形成;流式细胞术和Western blot检测证实PM2.5可以通过上调细胞色素C表达和活化cleaved caspase-9和cleaved caspase-3而诱导EA.hy926细胞凋亡.此外,用N-乙酰半胱氨酸抑制氧自由基生成可以抑制血管内皮细胞凋亡,提示PM2.5引起的细胞凋亡与氧化应激有关.结论:PM2.5可导致血管内皮细胞氧化应激水平增强和凋亡增加,这可能是其影响心血管系统功能的机制之一.
      Abstract:
      AIM:To study the toxicity of PM2.5 in the endothelial cells by investigating the induction of reactive oxygen species (ROS) and apoptosis in EA.hy926 cells exposed to PM2.5.METHODS: The endothelial cell line EA. hy926 was cultured in vitro and exposed to PM2.5 at different concentrations for 24 h.The cell viability was measured by CCK-8 assay and the generation of intracellular ROS was stained with DCFH-DA.The cell apoptosis was analyzed by flow cy-tometry with Annexin V-FITC/PI staining, and the protein levels of cytochrome C , caspase 9 and caspase 3 was detected by Western blot.RESULTS:After the treatment with PM2.5, the viability of the EA.hy926 cells was decreased markedly and the production of ROS was increased significantly .PM2.5 exposure upregulated the expression of cytoplasm cytochrome C and activated caspase-9 and caspase-3, resulting in the increase of the cell apoptosis significantly .The ROS generation was direct-ly involved in PM2.5-mediated endothelial cell apoptosis as N-acetyl-L-cysteine pretreatment abolished both ROS production and cell apoptosis induced by PM 2.5.CONCLUSION:PM2.5 induces oxidative stress and apoptosis in the vascular endo-thelial cells, which may be one of the mechanisms that PM2.5 influences the function of cardiovascular system .
      作者: 王芳 [1] 李岩 [2] 卢昕烁 [1] 杨杰波 [1] 刘方芳 [2] 余雪松 [1] 李明 [1]
      Author: WANG Fang [1] LI Yan [2] LU Xin-shuo [1] YANG Jie-bo [1] LIU Fang-fang [2] YU Xue-song [1] LI Ming [1]
      作者单位: 广东药科大学基础学院,广东广州,510006 广州中医药大学基础医学院,广东广州,510006
      刊 名: 中国病理生理杂志 ISTICPKU
      年,卷(期): 2017, 33(3)
      分类号: R363
      在线出版日期: 2017年4月10日
      基金项目: 广东省科技计划