PM2.5对血管内皮细胞氧化应激和凋亡的影响及机制

发布时间：2018-09-06 15:58

【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 to vascular endothelial cells (VECs) from the effects of atmospheric fine particles (PM2.5) on oxidative stress and apoptosis of vascular endothelial cells (VEC). Methods: cultured vascular endothelial cells (EA.hy926,) were exposed to different concentrations of PM2.5 for 24 h. The activity of the cells was measured by CCK-8 method, the production of oxygen free radicals was detected by DCFH-DA fluorescence labeling method, and the apoptosis rate was detected by flow cytometry. Then the expression of apoptosis-related protein cytochrome cleaved caspase-9 and cleaved caspase-3 was detected by Western blot assay. Results the results of 1% CCK-8 assay showed that PM2.5 had obvious toxicity to vascular endothelial cells. When the concentration was more than 25 mg/L, the activity of EA.hy926 cells was significantly decreased and the fluorescence staining of DCFH-DA was enhanced 24 h after exposure to PM2.5, indicating that there were a large number of oxygen free radicals formed in the cells. Flow cytometry and Western blot showed that PM2.5 could induce apoptosis of EA.hy926 cells by up-regulating cytochrome C expression and activating cleaved caspase-9 and cleaved caspase-3. In addition, the inhibition of oxygen free radical production by N-acetylcysteine can inhibit the apoptosis of vascular endothelial cells, suggesting that apoptosis induced by PM2.5 is related to oxidative stress. Conclusion the increase of oxidative stress and apoptosis of vascular endothelial cells induced by 1: PM2.5 may be one of the mechanisms that affect cardiovascular system function.
【作者单位】：广东药科大学基础学院;广州中医药大学基础医学院;
【基金】：广东省科技计划(No.2014A020212266;No.2016A020215156)
【分类号】：R54;X513

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