Effect and Mechanism of Azithromycin on Inflammatory Injury of Alveolar Epithelial Cells Induced by Lipopolysaccharide
To explore the regulatory effects of azithromycin on lipopolysaccharide(LPS)-induced proliferation,apoptosis and Janus kinase 2(JAK2)/signal transduction and transcription promoter 3(STAT3)signaling pathways in human alveolar epithelial cells,in vitro culture of human alveolar epithelial cells A549,were divided into blank group(no intervention),LPS group(10 μg/mL LPS treated for 24 h),low/medium/high concentration experimental group(10 μg/mL LPS+1,2,4 μg/mL azithromycin)and azithromycin group(10 μg/mL LPS+4 μg/mL azithromycin),inhibitor group(10 μg/mL LPS+50 μmol/L JAK2/STAT3 pathway inhibitor AG490),azithromycin+inhibitor group(10 μg/mL LPS+4 μg/mL azithromycin+50 μmol/L AG490)and azithromycin+activator group(10 μg/mL LPS+4 μg/mL azithromycin+0.5 μmol/L JAK2/STAT3 pathway acti-vator Colivelin).After 24 hours of intervention,enzyme-linked immunosorption assay(ELISA),cell counting kit-8(CCK-8),5-acetyney-2'deoxyuracil nucleoside(EdU),Hoechst 33258 staining and Western blotting(WB)were used for the expression levels of inflammatory factors interleukin-6(IL-6),interleukin-8(IL-8),tumor necrosis factor-α(TNF-α),cell viability,prolif-eration rate,apoptosis rate,Caspase-3 and Cyclin D1 and the expression levels of JAK2/STAT3 signaling pathway related pro-teins.The results show that:compared with blank group,the expression levels of inflammatory cytokines IL-6,IL-8 and TNF-α in LPS group significantly increased,and cell viability significantly decreased.Compared with LPS group,the expression levels of inflammatory cytokines IL-6,IL-8 and TNF-α in high-concentration experimental group significantly decreased,and cell vi-ability significantly increased.In this study,4 μg/mL azithromycin with significant difference from LPS group was selected as azithromycin group for follow-up experiments.Compared with blank group,the cell proliferation rate and Cyclin D1 protein ex-pression levels in LPS group significantly decreased,while the apoptosis rate,Caspase-3,p-JAK2 and p-STAT3 protein expres-sion levels significantly increased.Compared with LPS group,azithromycin group and inhibitor group significantly reversed the changes of the above indexes.Compared with azithromycin group,cell proliferation rate and Cyclin D1 protein expression lev-els in azithromycin+inhibitor group further significantly increased,while apoptosis rate,IL-6,IL-8,TNF-α,Caspase-3,p-JAK2 and p-STAT3 protein expression levels further significantly decreased.Azithromycin+activator group significantly reversed the changes of the above indexes.Azithromycin can reduce LPS-induced inflammatory damage of A549 cells by inhibiting JAK2/STAT3 signaling pathway,promote cell proliferation and inhibit apoptosis,and provide evidence for exploring azithromycin treatment of LPS induced acute lung injury.
国家科技期刊平台
NSTL
万方数据
