lncRNA NEAT1靶向miR-495-3p对LPS诱导的WI-38细胞损伤的影响  

Effects of lncRNA NEAT1 targeting miR-495-3p on WI-38 cell injury induced by LPS

作　　者：朱曦 郭赟 陆涛 钱俊 ZHU Xi;GUO Yun;LU Tao;QIAN Jun(Department of Critical Care Medicine,Affiliated Children's Hospital of Jiangnan University(Wuxi Children's Hospital),Wuxi 214023,China;Department of Respiratory Medicine,Affiliated Children's Hospital of Jiangnan University(Wuxi Children's Hospital),Wuxi 214023,China)

机构地区：[1]江南大学附属儿童医院塈无锡市儿童医院重症医学科,无锡214023 [2]江南大学附属儿童医院塈无锡市儿童医院呼吸内科,无锡214023

出　　处：《中国免疫学杂志》2025年第3期576-581,共6页Chinese Journal of Immunology

基　　金：江苏省医学创新团队(CXTDB2017016);江苏省无锡市科技局“太湖之光”科技攻关(基础研究)(K20221033)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)核富集转录体1(NEAT1)通过调控微小RNA-495-3p(miR-495-3p)对脂多糖(LPS)诱导的人胚肺细胞(WI-38)损伤的影响。方法:采用10μg/ml LPS处理WI-38细胞建立体外肺炎模型,采用实时荧光定量聚合酶链式反应(qPCR)检测WI-38细胞中NEAT1和miR-495-3p的表达水平,qPCR检测NEAT1和miR-495-3p的表达。采用双荧光素酶报告基因实验检测NEAT1和miR-495-3p的靶向关系。采用细胞计数试剂盒和流式细胞术分别检测敲低NEAT1对LPS诱导的WI-38细胞增殖活力和凋亡的影响,蛋白质免疫印迹试验检测Bcl-2、Bax和Cleaved caspase-3蛋白质表达。ELISA检测炎症因子IL-6、IL-8和IL-1β的含量。结果:LPS刺激能够上调WI-38中NEAT1的表达,抑制miR-495-3p的表达;双荧光素酶报告基因实验证实NEAT1和miR-495-3p存在靶向关系,且敲低NEAT1能够促进miR-495-3p的表达;LPS能够抑制WI-38细胞增殖促进细胞凋亡,促进炎症因子IL-6、IL-8和IL-1β的表达(P<0.05);敲低NEAT1能够抑制LPS诱导的WI-38细胞凋亡和炎症反应,促进细胞增殖。结论:敲低NEAT1通过靶向负调控miR-495-3p抑制LPS诱导的WI-38细胞凋亡和炎症反应。Objective:To investigate the effect of long non-coding RNA(lncRNA)nuclear enriched transcript 1(NEAT1)on lipopolysaccharide(LPS)-induced human embryonic lung cells(WI-38)injury by regulating microRNA-495-3p(miR-495-3p).Methods:WI-38 cells were treated with 10μg/ml LPS to establish an in vitro pneumonia model.Real-time fluorescent quantitative polymerase chain reaction(qPCR)was used to detect the expression levels of NEAT1 and miR-495-3p in WI-38 cells.The dual luciferase reporter assay was used to detect the targeting relationship between NEAT1 and miR-495-3p.Cell counting kit and flow cytometry were used to detect the effect of knockdown NEAT1 on the proliferation and apoptosis of WI-38 cells treated with LPS.Western blot was used to detect Bcl-2,Bax and Cleaved caspase-3 protein expressions.ELISA was used to detect the levels of inflammatory factors IL-6,IL-8 and IL-1β.Results:LPS stimulation could up-regulate the expression of NEAT1 and inhibit the expression of miR-495-3p in WI-38;NEAT1 directly targeted miR-495-3p,and NEAT1 knockdown could promote the expressions of miR-495-3p.Functionally,LPS inhibited the proliferation of WI-38 cells and promoted cell apoptosis and the expression of the inflammatory factors IL-6,IL-8 and IL-1β(P<0.05).Knockdown of NEAT1 could inhibit LPS-induced WI-38 cell apoptosis and inflammation,and promote cell proliferation.Conclusion:Knockdown of NEAT1 can inhibit LPS-induced apoptosis and inflammation of WI-38 cells by negatively regulating miR-495-3p.

关 键 词：lncRNA NEAT1 miR-495-3p 人胚肺细胞WI-38 凋亡 炎症 

分 类 号：R725.6[医药卫生—儿科]