Urban PM2.5 in Guangzhou induces apoptosis of PC-12 cells / 实用医学杂志

ABSTRACT

Objective To investigate the toxicity of urban PM2.5 in Guangzhou on PC-12 cells.Methods PM2.5 was collected in Guangzhou city.PC-12 cells were cultured in vitro and divided into several groups,including the control group,PM2.5 groups with different concentrations and NAC pretreated group (pretreaed with 10 μmol/mL NAC followed with 100 μg/mL PM2.5 exposure).Cells were incubated with indicated stimulator for 24 h,then cell viability was checked with cell counting Kit-8 assay,the level of intracellular ROS was labeled using H2DCFDA fluorescence probe,cell apoptosis was measured by the flow cytometry and the expression of apoptosis-related proteins,including Cytochrome C,Caspase 9,Caspase 3 and PARP,were detected by Western Blot assay.Results PM2.5 has a strong toxicity on PC-12 cells when its concentration is over 25 μg/mL.After exposure for 24 h,the cell viability was markedly decreased.The results of flow cytometry and Western blot assay showed that PM2.5 enhanced the apoptosis of PC-12 cells with the upregulatios of Cytochrome C,Caspase 9,Caspase 3 and PARP.Pretreatment with NAC could significantly diminish PM2.5-induced PC-12 cell toxicity,decreased ROS generation and apoptosis of PC-12 cells,with the down regulations of apoptosis-related proteins.Conclusion PM2.5 can cause apoptosis of PC-12 cells by inducing oxidative stress,upregulating the Cytochrome C expression and activating Caspase9/3,which may be one of the mechanisms underlying PM2.5-induced neurotoxicity.