The Effect Of PM2.5 In Taiyuan On JAK/ STAT Signaling Pathway Of Human Bronchial Epithelial Cells

Part1 The Effect of PM2.5 on Oxidative Stress-JAK2/STAT3 Signaling Pathway of Human Bronchial Epithelial Cells Objective:To investigate the regulation of oxidative stress-JAK2/STAT3 signaling pathway on cytokines of human bronchial epithelial cells induced by PM2.5, to provide new ideas for further studying of the pathogenesis of respiratory diseases. Methods:Bronchial epithelial cells 16 HBE were cultured using traditional invasive methods. There were the following groups: control group, N-acetyl-L-cysteine(NAC,5 mmol/L) group, 6 μmol/L AG490 group, 50 μg/ml PM2.5 group, 100 μg/ml PM2.5 group,50 μg/ml PM2.5+5 mmol/L NAC group, 100 μg/ml PM2.5+5 mmol/L NAC group, 100 μg/ml PM2.5+6 μmol/L AG490. Intracellular ROS levels, the ratio of GSH/GSSG, the gene and protein expression of JAK2 and STAT3 and the level of IL-6, IL-8 and the protein expression of COX-2 were detected after all groups were exposed for 12 h and 24 h. Results:After 12 h and 24 h exposure, intracellular ROS levels, the gene and protein expression of JAK2 and STAT3 in 50 μg/ml PM2.5, 100 μg/ml PM2.5 exposure group were higher than the control group, however the ratio of GSH/GSSG appeared the opposite result. Intracellular ROS levels, the gene and protein expression of JAK2 and STAT3 in PM2.5 + NAC protection group were lower than the respective PM2.5 exposure group, but the ratio of GSH/GSSG was higher than the respective PM2.5 exposure group, the differences were statistically significant(P <0.05). Cellular IL-6 and IL-8 levels and the protein expression of COX-2 in 100 μg/ml PM2.5 group were higher than control group, 5mmol/L NAC group, 6μmol/L AG490 group, 100μg/ml PM2.5 +5mmol/L NACgroup, 100 μg/ml PM2.5+6μmol/L AG490 group, the differences were statistically significant(P<0.05). Conclusions:PM2.5 could cause oxidative damage on bronchial epithelial cells, and oxidative stress could regulate JAK/STAT signaling pathway and relative cytokines of epithelial.Part2 The Effect of PM2.5 on IL4R-JAK3/STAT6-Eotaxin Signaling Pathway of Human Bronchial Epithelial Cells Objective:To investigate the regulation of JAK3/STAT6 signaling pathway on cytokines of human bronchial epithelial cells induced by PM2.5, and to provide new ideas for further studying of the pathogenesis of respiratory diseases.Methods:Bronchial epithelial cells 16 HBE were cultured using traditional invasive methods. There were the following groups: control group, 50 μg/ml PM2.5 group, 100 μg/ml PM2.5 group, 10ng/ml IL-4 group, 50μg/ml PM2.5 + 10ng/ml IL-4 group, 100μg/ml PM2.5 + 10ng/ml IL-4 group, 10ng/ml IL-4+6 μmol/L AG490 group, 10ng/ml IL-4+6μmol/L AG490+100μg/ml PM2.5 group, The protein expression of IL-4Rα,the gene and protein expression of JAK3 and STAT6 and the intracellular eotaxin was determined after all groups were exposed for 12 h and 24 h. Results:After 12 h and 24 h exposure, the protein expression of IL-4Rα in 50 μg/ml PM2.5, 100 μg/ml PM2.5, 10ng/ml IL-4, 50μg/ml PM2.5 + 10ng/ml IL-4, 100μg/ml PM2.5 + 10ng/ml IL-4 groups were higher than the control group. The protein expression of IL-4Rα in 50μg/ml PM2.5 + 10ng/ml IL-4, 100μg/ml PM2.5 + 10ng/ml IL-4 groups were higher than the 10ng/ml IL-4 group,and the differences were statistically significant(P <0.05). The gene and protein expression of JAK3 and STAT6 in 50μg/ml PM2.5 + 10ng/ml IL-4, 100μg/ml PM2.5 + 10ng/ml IL-4 groups were higher than the control group and10ng/ml IL-4 group,the differences were statistically significant(P <0.05). After 12 h and 24 h exposure, intracellular eotaxin levels in 100μg/ml PM2.5+10ng/ml IL-4 exposure group were higher than the control group, 10ng/ml IL-4 group, 10ng/ml IL-4+6 μmol/L AG490 group and 10ng/ml IL-4+6μmol/L AG490+100μg/ml PM2.5 group, the differences were statistically significant(P <0.05). Conclusions:IL-4R-JAK3 / STAT6 signaling pathway involves in the regulation of eotaxin of bronchial epithelial cells induced by PM2.5.