CXCL2基因在LPS诱导的支气管上皮细胞炎症反应中的作用  

Role of CXCL2 gene in LPS-induced inflammation of bronchial epithelial cells

作　　者：钟甜 郭璐[1] 蒋才玉 彭夏莹 邹俊[1] ZHONG Tian;GUO Lu;JIANG Cai-Yu;PENG Xia-Ying;ZOU Jun(Sichuan Academy of Medical Sciences·Sichuan Provincial People's Hospital,Respiratory and Critical Care Medicine,Chengdu 610072,China)

机构地区：[1]四川省医学科学院·四川省人民医院呼吸与危重症医学科,成都610072

出　　处：《中国免疫学杂志》2021年第15期1804-1808,共5页Chinese Journal of Immunology

基　　金：四川省卫生和计划生育委员会科研课题(17PJ369);四川省医学科学院·四川省人民医院青年人才基金项目(2016QN07)。

摘　　要：目的:探讨CXCL2基因对脂多糖(LPS)诱导的人支气管上皮细胞(16HBE)炎症反应的影响及机制。方法:体外培养16HBE细胞,采用50μg/ml的LPS干预16HBE细胞,构建LPS诱导的16HBE细胞炎症反应模型,实验分为未处理对照组(Control组)、LPS模型组(LPS组)、转染siRNA NC+LPS处理组(si-NC+LPS组)和转染CXCL2 siRNA+LPS处理组(si-CXCL2+LPS组)。分别采用qPCR和Western blot分析CXCL2 mRNA和蛋白的表达情况,MTT法检测16HBE细胞增殖活力,ELISA检测TNF-α、IL-6和IL-1β含量,流式细胞术检测16HBE细胞凋亡率,Western blot检测JAK2和STAT3的磷酸化水平。结果:与Control组相比,LPS组16HBE细胞中CXCL2 mRNA和蛋白的表达明显上调,细胞增殖活力明显降低,TNF-α、IL-6和IL-1β含量明显增多,细胞凋亡率明显升高,JAK2和STAT3磷酸化水平明显升高,差异有统计学意义(P<0.05);与LPS组相比,si-CXCL2+LPS组16HBE细胞中CXCL2 mRNA、CXCL2蛋白、TNF-α、IL-6和IL-1β的含量、JAK2和STAT3的磷酸化水平以及细胞凋亡率均明显降低,细胞增殖活力明显升高,差异有统计学意义(P<0.05);LPS组与si-NC+LPS组间以上各指标差异无统计学意义(P>0.05)。结论:沉默CXCL2基因能够减轻LPS诱导的支气管上皮细胞炎症反应,其作用机制可能与抑制JAK2/STAT3信号通路激活有关。Objective:To investigate the effect and mechanism of CXCL2 gene on lipopolysaccharide(LPS)-induced inflammation of human bronchial epithelial cells(16HBE).Methods:16HBE cells were cultured in vitro,and 50μg/ml LPS was used to intervene in 16HBE cells to construct an LPS-induced model of 16HBE cell inflammation.Experiment was divided into untreated control group(Control group),LPS model group(LPS group),transfected siRNA NC+LPS treatment group(si-NC+LPS group)and transfected CXCL2 siRNA+LPS treatment group(si-CXCL2+LPS group).qPCR and Western blot were used to analyze expression of CXCL2 mRNA and protein.MTT was used to detect proliferation activity of 16HBE cells,contents of TNF-α,IL-6 and IL-1βwere detected by ELISA,flow cytometry to detect 16HBE cell apoptosis rate,Western blot was used to detect phosphorylation levels JAK2 and STAT3.Results:Compared with Control group,expression of CXCL2 mRNA and protein in 16HBE cells of LPS group was significantly increased,the cell proliferation activity was significantly reduced,content of TNF-α,IL-6 and IL-1βwas significantly increased,and the apoptosis rate was significantly increased.JAK2 and STAT3 phosphorylation levels were significantly increased,the difference was statistically significant(P<0.05);compared with LPS group,CXCL2 mRNA,CXCL2 protein,TNF-α,IL-6 in 16HBE cells of si-CXCL2+LPS group and IL-1βcontent,JAK2 and STAT3 phosphorylation levels and apoptosis rate were significantly reduced,and cell proliferation activity was significantly increased,the difference was statistically significant(P<0.05);in LPS group and si-NC+LPS group,there was no statistically significant difference between the above indexes(P>0.05).Conclusion:Silencing CXCL2 gene can reduce LPS-induced bronchial epithelial inflammatory response,and its mechanism may be related to the inhibition of JAK2/STAT3 signaling pathway activation.

关 键 词：CXCL2基因 脂多糖 支气管上皮细胞 炎症反应 JAK2/STAT3信号通路 

分 类 号：R73[医药卫生—肿瘤]