PM2.5对支气管上皮细胞凋亡基因表达水平的影响

The effect of PM2.5 on apoptosis gene expression in HBE cells

目的探讨PM2.5对支气管上皮(HBE)细胞凋亡基因表达的影响。方法用PM2.5低剂量10μg/m L和高剂量50μg/mL对HBE细胞染毒,用未染毒的细胞作为对照组,10μg/mL的Cr6+为阳性对照组,应用实时荧光定量聚合酶链式反应(qRTPCR)检测凋亡基因Caspase-3、Caspase-8、Caspase-9、Bax、Bcl-2的mRNA表达水平变化情况,Western blot检测凋亡基因的蛋白质表达变化情况。结果 qRT-PCR结果显示,与对照组比较,PM2.510μg/mL和50μg/mL染毒以及阳性对照组染毒后,Caspase-3基因表达量分别升高193.0%、340.0%、214.0%,Caspase-8基因表达分别升高50.0%、84.0%、71.0%,Caspase-9基因表达分别升高94.0%、177.0%、117.0%,Bax基因表达分别升高8.0%、38.0%、57.0%,Bcl-2基因表达水平分别下降26.0%、46.0%、57.0%。Western blot结果显示,与对照组比较,Caspase-3蛋白在PM2.510μg/mL和50μg/mL染毒和Cr6+染毒后表达水平分别升高22.4%、48.6%、21.8%,Caspase-8蛋白表达水平分别升高54.7%、77.8%、64.3%,Caspase-9蛋白表达水平分别升高16.1%、82.7%、25.1%,Bax蛋白表达水平分别升高54.7%、90.4%、42.8%,Bcl-2蛋白表达量分别下降20.5%、41.8%、27.6%。结论 PM2.5染毒引起HBE细胞中凋亡基因表达升高,抑凋亡基因表达下降,提示雾霾对凋亡基因表达水平具有明显影响。

Objective To understand the effect of particulate matter 2.5(PM2.5)on apoptosis gene expression in human bronchial epithelial(HBE)cells.Methods HBE cells were treated with PM2.5 for 24 h at 10μg/mL and 50μg/mL,additionally,cells were treated with blank as negative control,10μg/mL Cr6+as a positive control.After the treatment,m RNA expression of apoptosis genes including Caspase-3,Caspase-8,Caspase-9,Bax,and Bcl-2 were detected by fluorescent quantitative RT-PCR(qRTPCR),the protein expression of apoptosis genes were detected with Western blot.Results The qRT-PCR showed that compared with control group,after exposure to PM2.510μg/mL,50μg/mL and positive control,the m RNA expression level of Caspase-3 gene increased by 193.0%,340.0%and 214.0%,Caspase-8 expression level increased by 50.0%,84.0%and 71.0%,Caspase-9 expression level increased by 94.0%,177.0%and 117.0%,Bax expression level increased by 8.0%,38.0%and 57.0%,Bcl-2 expression level decreased by 26.0%,46.0%,and 57.0%,respectively.Western blot analysis showed that compared with the control group,after exposure to PM2.510μg/mL,50μg/mL and positive control,the expression level of Caspase-3 protein increased by 22.4%,48.6%and 21.8%,Caspase-8 protein increased by 54.7%,77.8%and 64.3%,Caspase-9 protein increased by 16.1%,82.7%and 25.1%,Bax protein increased by 54.7%,90.4%and 42.8%,Bcl-2 protein decreased by 20.5%,41.8%and 27.6%,respectively.Conclusion PM2.5 exposure can increase the expression of apoptotic genes and decrease the expression.