【摘要】：目的:研究細(xì)顆粒物(PM2.5)對(duì)PC-12細(xì)胞凋亡的影響及其信號(hào)途徑。方法:從廣州城區(qū)采集PM2.5,對(duì)PC-12細(xì)胞染毒,設(shè)對(duì)照組、不同濃度PM2.5組和N-乙�；腚装彼�(NAC)處理組,染毒24 h后用CCK-8法檢測細(xì)胞存活率,H2-DCFDA探針標(biāo)記法測定細(xì)胞內(nèi)活性氧(ROS)水平,流式細(xì)胞術(shù)檢測細(xì)胞凋亡情況,Western blot法檢測凋亡相關(guān)蛋白的表達(dá)變化。結(jié)果:當(dāng)濃度大于25μg/m L時(shí)PM2.5對(duì)PC-12細(xì)胞有較強(qiáng)的毒性,細(xì)胞存活率明顯下降;用25、50和100μg/m L的PM2.5染毒24 h細(xì)胞內(nèi)可見大量綠色熒光,說明ROS生成增多;流式細(xì)胞術(shù)和Western blot結(jié)果顯示PM2.5染毒組細(xì)胞凋亡率明顯增高,并伴隨有Cytochrome C表達(dá)上調(diào)、Caspase 9和Caspase 3激活及PARP增多;加入抗氧化劑NAC可以抑制氧自由基生成,使細(xì)胞凋亡減少并下調(diào)相關(guān)分子的表達(dá)或激活。結(jié)論:PM2.5可通過誘導(dǎo)氧化應(yīng)激,上調(diào)Cytochrome C表達(dá),激活Caspase 9/3誘導(dǎo)PC-12細(xì)胞凋亡,這可能是PM2.5影響PC-12細(xì)胞功能的機(jī)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.
【作者單位】： 廣東藥科大學(xué)基礎(chǔ)學(xué)院;廣州中醫(yī)藥大學(xué)基礎(chǔ)醫(yī)學(xué)院;廣東藥科大學(xué)醫(yī)藥化工學(xué)院;
【基金】：廣東省科技計(jì)劃項(xiàng)目(編號(hào):2014A020212266,2016A020215156) 廣州中醫(yī)藥大學(xué)薪火計(jì)劃項(xiàng)目(編號(hào):XH20150101);廣州中醫(yī)藥大學(xué)青年英才項(xiàng)目(編號(hào):QNYC20140102)
【分類號(hào)】：R122

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