The Damage Effects Of Atmospheric PM_2.5 On Human Bronchial Epithelial Cells BEAS-2B In Shijiazhuang

Objective:To study the damage effects of atmospheric fine particulate matters PM_2.5 on the human bronchial epithelial cells BEAS-2B,and its possible mechanism on respiratory injury;to provide a scientific basis for respiratory diseases caused by atmospheric particulate matters,and an important clue for subsequent research.Methods:PM_2.5 were collected during the heating period of Shijiazhuang city,eluted with ultrapure water,and collected after freeze-drying by vacuum dryer.Then they were prepared into different concentrations in PBS.The BEAS-2B cells were cultured in different PM_2.5 concentrations of 0,100,200,and 400?g/ml respectively during a period of 6,24,and 48h respectively.The viability of cells was determined by MTT assay,and LDH leakage was determined by micro-enzyme method,and cell proliferation was determined by 5-BrdU incorporation method to evaluate the comprehensive toxicity of PM_2.5 on cells;intracellular ROS content,MDA production,SOD activity,and CAT activity were determined to evaluate the oxidative damage effect of PM_2.5on cells;the content of IL-1?,IL-10,IL-12p70,and TNF-?were determined by flow cytometry to evaluate the inflammatory damage effect of PM_2.5 on cells.SPSS 21.0 software was used for statistical analysis;multiple groups were compared through one-way ANOVA analysis;multiple exposure concentration groups compared with 0?g/ml group,and 24,48h exposure time group compared with 6h group were analyzed through Dunnett-t test.Results:1.The comprehensive toxity effect of PM_2.5 on cells:Compared with the0?g/ml group at the same exposure time,the survival rate of cells gradually decreased,and the LDH leakage increased,and the proportion of proliferating cells decreased in 100,200 and 400?g/ml group,the difference was statistically significant?P<0.05?;compared with the 6h group at the same exposure concentration,the survival rate of cells gradually decreased,and the LDH leakage increased,and the proportion of proliferating cells decreased in24,48h group,the difference was statistically significant?P<0.05?.2.The oxidative damage effect of PM_2.5 on cells:Compared with the0?g/ml group at the same exposure time,intracellular ROS content and MDA production gradually increased,and SOD and CAT activity gradually decreased in 100,200 and 400?g/ml group,the difference was statistically significant?P<0.05?;compared with the 6h group at the same exposure concentration,intracellular ROS content and MDA production gradually increased,and SOD and CAT activity gradually decreased in 24,48h group,the difference was statistically significant?P<0.05?.3.The inflammatory damage effect of PM_2.5 on cells:Compared with the0?g/ml group at the same exposure time,the cytokines content of IL-1?,IL-10,IL-12p70 and TNF-?gradually increased in 100,200 and 400?g/ml group,the difference was statistically significant?P<0.05?.Compared with the 6h group at the same exposure concentration,the cytokines content of IL-1?,IL-10,IL-12p70 and TNF-?didn't increase.Conclusions:Atmospheric fine particles PM_2.5 produced cytotoxic effects on BEAS-2B cells and induced oxidative stress and inflammatory responses in respiratory cells,and the damage effects gradually worsen with the PM_2.5 exposure concentration increasing and exposure time prolonging.Oxidative stress and inflammatory reaction may be the main mechanism of PM_2.5 injury on respiratory system.