Regulation Of Ca²⁺JAK1-STAT1 Signaling Pathway On Inflammatory Mediators Changes Of 16HBE Cells Induced By PM2.5

Objective:To explore the impact of Ca2+-JAK1-STAT1 signaling pathway on inflammatory mediators of human bronchial epithelial cells induced by PM2.5, to provide reference for the immune mechanism of asthma caused by PM2.5. Methods:Normal saline control group,100μg/ml heparin group,50μg/ml PM2.5 group,100μg/ml PM2.5 group,50μg/ml PM2.5+100μg/ml heparin group,100μg/ml PM2.5+100μg/ml heparin group were set. After the 16 HBE cells were exposed for 3h, 6h and 24 h, the changes of intracellular [Ca2+]i were detected by FCM and the gene and protein expression of JAK1 and STAT1 by QT-PCR and Western blotting. Normal saline control group,100μg/ml heparin group,10μM Cur group,50μg/ml PM2.5 group,100μg/ml PM2.5 group,50μg/ml PM2.5+100μg/ml heparin group,100μg/ml PM2.5+100μg/ml heparin group,50μg/ml PM2.5 +10μM Cur group,100μg/ml PM2.5+10μM Cur group,50μg/ml PM2.5 +100μg/ml heparin+10μM Cur group,100μg/ml PM2.5+100μg/ml heparin+10μM Cur group were set respectively. After the cells were exposed for 3h,6h and 24 h,the levels of TNF-α,ICAM-1 and HMGB1 were detected by ELISA. Results:1.After 6h and 24 h exposure, the intracellular [Ca2+]i in 50μg/ml PM2.5 and 100μg/ml PM2.5 groups was higher than the saline group and the differences were statistically significant(P<0.05). The intracellular [Ca2+]i in PM2.5+100μg/ml heparin groups were lower than alone PM2.5 groups.2. After 3h,6h and 24 h exposure, the gene and protein expression of JAK1 and STAT1 in 50μg/ml PM2.5 and 100μg/ml PM2.5 groups were higher than the saline group and the differences were statistically significant(P<0.05).The gene and protein expression of JAK1 in PM2.5+100μg/ml heparin groups were lower than alone PM2.5 groups. The gene and protein expression of STAT1 in PM2.5+100μg/ml heparin groups were lower than alone PM2.5 groups after 3h and 6h exposure,while the increasing tendency was presented after 24 h exposure.3. The levels of TNF-α, ICAM-1 and HMGB1 in 50μg/ml PM2.5 and 100μg/ml PM2.5 groups were higher than the saline group,100μg/ml heparin group and 10μM Cur group and the differences were statistically significant(P<0.05). Conclusion:PM2.5 could increase the levels of TNF-α, ICAM-1 and HMGB1 of human bronchial epithelial cells by Ca2+-JAK1-STAT1 signalling pathway.