Effects of PM2.5 on Barrier Function,Oxidative Stress Response and Autophagy of Human Corneal Epithelial Cell
Objective:To investigate the effects of PM2.5 on barrier function,oxidative stress response and autophagy of human corneal epithelial cell and its molecular mechanism.Methods:Human corneal epithelial cells(HCEC)were treated with different concen-trations of PM2.5 for 24 h.The cell proliferation level was detected by CCK8.Intracellular reactive oxygen species(ROS)were detected by fluorescent labeling.The levels of IL-6,IL-8 and TNF-α were detected by ELISA kit.The changes of TEER in HCEC cells were de-tected by transmembrane resistance assay(TEER).The protein expression levels of Beclin1,LC3 Ⅱ/Ⅰ,Claudin1,Occludin and ZO-1 were detected by Western blot.Then HCEC cells were divided into control group,PM2.5 group,NAC group and PM2.5+NAC group,and the cells of all groups were cultured for 24 h.Cell proliferation level,ROS,inflammatory factors,transmembrane resistance and Western blot were subsequently detected.Results:Compared with 0 μg/mL,the relative proliferation level,TEER value,and the levels of Claudin1,ZO-1 and Occludin protein of HCEC cells treated with 40 µg/mL,80 µg/mL,160 μg/mL and 200 µg/mL PM2.5 were de-creased(P＜0.05),and the levels of IL-6,IL-8 and TNF-α in supernatant were increased(P＜0.05).The level of ROS and the protein levels of Beclin-1 and LC3 Ⅱ/Ⅰ in HCEC cells were increased(P＜0.05).Compared with the PM2.5 group,the relative proliferation level,TEER value,and the levels of Claudin1,ZO-1 and Occludin protein of HCEC cells in PM2.5+NAC group was increased(P＜0.05),the levels of IL-6,IL-8 and TNF-α in supernatant were decreased(P＜0.05),and the level of ROS and the protein levels of Beclin-1 and LC3Ⅱ/Ⅰ in HCEC cells were decreased(P＜0.05).Compared with the NAC group,the relative proliferation level,TEER value,and the lev-els of Claudin1,ZO-1 and Occludin protein of HCEC cells in PM2.5+NAC group was decreased(P＜0.05),the levels of IL-6,IL-8 and TNF-α in supernatant were increased(P＜0.05),and the level of ROS and the protein levels of Beclin-1 and LC3 Ⅱ/Ⅰ in HCEC cells were increased(P＜0.05).Conclusions:PM2.5 damages the barrier function of human corneal epithelial cells and promotes autophagy by inducing inflammation and oxidative stress responses.
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