广州城区大气细颗粒物对PC-12细胞凋亡的影响
发布时间：2018-07-29 11:45
【摘要】：目的:研究细颗粒物(PM2.5)对PC-12细胞凋亡的影响及其信号途径。方法:从广州城区采集PM2.5,对PC-12细胞染毒,设对照组、不同浓度PM2.5组和N-乙酰基半胱氨酸(NAC)处理组,染毒24 h后用CCK-8法检测细胞存活率,H2-DCFDA探针标记法测定细胞内活性氧(ROS)水平,流式细胞术检测细胞凋亡情况,Western blot法检测凋亡相关蛋白的表达变化。结果:当浓度大于25μg/m L时PM2.5对PC-12细胞有较强的毒性,细胞存活率明显下降;用25、50和100μg/m L的PM2.5染毒24 h细胞内可见大量绿色荧光,说明ROS生成增多;流式细胞术和Western blot结果显示PM2.5染毒组细胞凋亡率明显增高,并伴随有Cytochrome C表达上调、Caspase 9和Caspase 3激活及PARP增多;加入抗氧化剂NAC可以抑制氧自由基生成,使细胞凋亡减少并下调相关分子的表达或激活。结论:PM2.5可通过诱导氧化应激,上调Cytochrome C表达,激活Caspase 9/3诱导PC-12细胞凋亡,这可能是PM2.5影响PC-12细胞功能的机制之一。
[Abstract]:Aim: to study the effect of fine particles (PM2.5) on apoptosis of PC-12 cells and its signal pathway. Methods: PM2.5 was collected from Guangzhou urban area and treated with PC-12 cells. The cells were treated with different concentrations of PM2.5 and N-acetylcysteine (NAC). After 24 hours of exposure, the cell survival rate was detected by CCK-8 method and the intracellular reactive oxygen species (Ros) (ROS) levels were detected by H2-DCFDA probe labeling method. Apoptosis was detected by flow cytometry and the expression of apoptosis-related protein was detected by Western blot. Results: when the concentration of PM2.5 was more than 25 渭 g / mL, PM2.5 was more toxic to PC-12 cells, and the cell survival rate was significantly decreased, and a large amount of green fluorescence could be observed in the cells exposed to 25 渭 g / mL and 100 渭 g / mL PM2.5 for 24 h, indicating that the production of ROS increased. The results of flow cytometry and Western blot showed that the apoptosis rate of PM2.5 exposed group was significantly increased, accompanied by up-regulation of Cytochrome C expression, activation of caspase 9 and Caspase 3, and increase of PARP, and the addition of antioxidant NAC could inhibit the production of oxygen free radicals. Reduce apoptosis and down-regulate the expression or activation of related molecules. Conclusion the PC-12 cells apoptosis induced by Caspase 9 / 3 could be induced by oxidative stress, up-regulated expression of Cytochrome C and activation of Caspase 9 / 3, which may be one of the mechanisms by which PM2.5 affects the function of PC-12 cells.
【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;广东药科大学医药化工学院;
【基金】：广东省科技计划项目(编号:2014A020212266,2016A020215156) 广州中医药大学薪火计划项目(编号:XH20150101);广州中医药大学青年英才项目(编号:QNYC20140102)
【分类号】：R122
[Abstract]:Aim: to study the effect of fine particles (PM2.5) on apoptosis of PC-12 cells and its signal pathway. Methods: PM2.5 was collected from Guangzhou urban area and treated with PC-12 cells. The cells were treated with different concentrations of PM2.5 and N-acetylcysteine (NAC). After 24 hours of exposure, the cell survival rate was detected by CCK-8 method and the intracellular reactive oxygen species (Ros) (ROS) levels were detected by H2-DCFDA probe labeling method. Apoptosis was detected by flow cytometry and the expression of apoptosis-related protein was detected by Western blot. Results: when the concentration of PM2.5 was more than 25 渭 g / mL, PM2.5 was more toxic to PC-12 cells, and the cell survival rate was significantly decreased, and a large amount of green fluorescence could be observed in the cells exposed to 25 渭 g / mL and 100 渭 g / mL PM2.5 for 24 h, indicating that the production of ROS increased. The results of flow cytometry and Western blot showed that the apoptosis rate of PM2.5 exposed group was significantly increased, accompanied by up-regulation of Cytochrome C expression, activation of caspase 9 and Caspase 3, and increase of PARP, and the addition of antioxidant NAC could inhibit the production of oxygen free radicals. Reduce apoptosis and down-regulate the expression or activation of related molecules. Conclusion the PC-12 cells apoptosis induced by Caspase 9 / 3 could be induced by oxidative stress, up-regulated expression of Cytochrome C and activation of Caspase 9 / 3, which may be one of the mechanisms by which PM2.5 affects the function of PC-12 cells.
【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;广东药科大学医药化工学院;
【基金】：广东省科技计划项目(编号:2014A020212266,2016A020215156) 广州中医药大学薪火计划项目(编号:XH20150101);广州中医药大学青年英才项目(编号:QNYC20140102)
【分类号】：R122
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