摘要
目的探讨IL-6通过调控酪氨酸蛋白激酶2(JAK2)/信号传导与转录激活因子3(STAT3)信号通路减轻急性肺损伤(ALI)的机制。方法将人肺癌细胞A549细胞分为空白对照组、脂多糖(LPS)组、LPS+IL-6过表达组、LPS+IL-6干扰组、LPS+空载组。空白对照组细胞正常培养,LPS组LPS处理细胞,LPS+IL-6过表达组LPS处理细胞后转染IL-6过表达质粒,LPS+IL-6干扰组LPS处理细胞后转染IL-6干扰质粒,LPS+空载组LPS处理细胞后转染空载质粒。采用细胞计数试剂盒法检测细胞增殖率,流式细胞术检测细胞凋亡率和活性氧(ROS)水平,试剂盒检测丙二醛(MDA)和超氧化物歧化酶(SOD)水平,ELISA法检测炎性因子TNF-α、IL-6、IL-1β水平,qRT-PCR法检测IL-6、JAK2、STAT3 mRNA表达水平,Western blot法检测磷酸化JAK2(p-JAK2)/JAK2、磷酸化STAT3(p-STAT3)/STAT3蛋白表达水平。结果与空白对照组比较,过表达IL-6可增加LPS诱导的ALI细胞凋亡率、ROS、MDA、TNF-α、IL-6、IL-1β水平,JAK2、STAT3 mRNA表达水平,p-JAK2、p-STAT3蛋白表达水平,降低细胞增殖率和SOD水平;干扰IL-6则相反。结论IL-6可通过抑制JAK2/STAT3信号通路的激活,降低氧化应激和炎症反应,从而减轻ALI。
Objective To explore the mechanism by which IL-6 attenuates acute lung injury(ALI)by regulating janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway.Methods Human lung cancer cell line A549 cells were divided into blank control group(normally cultured),lipopolysaccharide(LPS)group(treated with LPS),LPS+IL-6 overexpression group(treated with LPS and transfected with IL-6 overexpression plasmid),LPS+IL-6 interference group(treated with LPS and transfected with IL-6 interference plasmid),and LPS+empty load group(treated with LPS and transfected with empty plasmid).Cell counting kit-8(CCK-8)assay was used to detect the cell proliferation rate.Rate of apoptosis and reactive oxygen species(ROS)content were measured by flow cytometry.The contents of malondialdehyde(MDA)and superoxide dismutase(SOD)were detected by the kit.The levels of inflammatory factors of TNF-α,IL-6 and IL-1βwere detected by ELISA.The gene expression levels of IL-6,JAK2 and STAT3 were detected by qRT-PCR.The protein expression levels of phosphorylated JAK2(p-JAK2)/JAK2,and phosphorylated STAT3(p-STAT3)/STAT3 were determined by Western blot assay.Results Compared to the blank control group,overexpression of IL-6 increased the apoptosis rates,the contents of ROS,MDA,TNF-α,IL-6,IL-1β,the gene expression levels of JAK2 and STAT3,and the protein expression levels of p-JAK2 and p-STAT3 in ALI cells induced by LPS,and decreased the proliferation rate and the content of SOD;while results of interfering with IL-6 were the opposite.Conclusion IL-6 can inhibit the activation of JAK2/STAT3 signaling pathway,and reduce oxidative stress and inflammatory reaction,and thus attenuates ALI.
作者
周斌
万少兵
王瑛
余平
典万康
周莹
周琴
ZHOU Bin;WAN Shaobing;WANG Ying;YU Ping;DIAN Wankang;ZHOU Ying;ZHOU Qin(Department of Emergency,Wuhan Third Hospital,Wuhan 430060,China)
出处
《浙江医学》
CAS
2024年第2期131-138,I0004,共9页
Zhejiang Medical Journal
基金
武汉市卫健委基金项目(WX21B32)。
作者简介
通信作者:周斌,E-mail:361635695@qq.com。
