lncRNA NEAT1对脂多糖所致肺泡巨噬细胞NF-κB活化的影响  

Effect of lncRNA NEAT1 on NF-κB activation in alveolar macrophages induced by lipopolysaccharide

作　　者：黄舒颖 张诚[2] 黄自坤[3] 罗清[3] 卿城[4] HUANG Shuying;ZHANG Cheng;HUANG Zikun(Department of Obstetrics and Gynecology,The First Affiliated Hospital of Nanchang University,Nanchang 330006,China;Department of Ultrasonography,The First Affiliated Hospital of Nanchang University,Nanchang 330006,China;Department of Clinical Laboratory,The First Affiliated Hospital of Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第一附属医院妇产科,南昌330006 [2]南昌大学第一附属医院超声科,南昌330006 [3]南昌大学第一附属医院检验科,南昌330006 [4]南昌大学第一附属医院重症医学科,南昌330006 [5]江西卫生职业学院助产系,南昌330052

出　　处：《江西医药》2020年第3期259-262,共4页Jiangxi Medical Journal

基　　金：江西省自然科学基金,编号20171BAB205028;江西省教育厅科学技术研究项目,编号GJJ160120;江西省卫生计生委科技计划,编号20185084,20204780。

摘　　要：目的检测沉默lncRNA NEAT1对LPS所诱导的NR8383肺泡巨噬细胞炎症因子IL-6分泌,NF-κB核移位的影响,探讨NEAT1调控肺泡巨噬细胞炎症反应的可能机制。方法NR8383肺泡巨噬细胞分为对照组,LPS(10 mg/L)组,LPS(10 mg/L)+siNC组,LPS(10 mg/L)+siNEAT1组。分别在6h后收集各组细胞和培养上清,RT-qPCR检测细胞内IL-6 mRNA和NEAT1表达,ELISA检测上清中细胞因子IL-6蛋白表达分泌,Western blot实验检测细胞核内NF-κB p65表达。结果LPS刺激NR8383肺泡巨噬细胞后,与对照组相比,细胞内NEAT1和IL-6 mRNA表达上调,培养上清中IL-6蛋白分泌增加(均P<0.05);siRNA沉默NEAT1后,巨噬细胞内NEAT1表达下调;与LPS组比较,siNEAT1组培养上清中IL-6分泌减少,NF-κB p65蛋白在细胞核内表达减少(均P<0.05)。结论肺泡巨噬细胞沉默lncRNA NEAT1可抑制IL-6分泌和NF-κB核转位,推测NEAT1通过此机制减轻NR8383肺泡巨噬细胞炎症反应。Objective To investigate the possible mechanism of NEAT1 regulating inflammatory response of alveolar macrophages.Methods NR8383 alveolar macrophages were divided into control group,LPS(10 mg/L)group,LPS(10 mg/L)+siNC group,LPS(10 mg/L)+siNEAT1 group.After 6h,the cells in each group were collected and supernatants were cultured.The expressions of IL-6 mRNA and NEAT1 in the cells were detected by RT-qPCR,the secretion of cytokine IL-6 protein was detected by ELISA,and the expression of NF-κB p65 in the nucleus was detected by Western blot.Results After LPS stimulated NR8383 alveolar macrophages,compared with the control group,intracellular NEAT1 and IL-6 mRNA expressions were up-regulated,and IL-6 protein secretion in culture supernatant was increased(all P<0.05).After silencing NEAT1 by siRNA,NEAT1 expression in macrophages was down-regulated.Compared with the LPS group,the production of IL-6 and the expression of NF-κB p65 decreased in the siNEAT1 group(all P<0.05).Conclusion Silencing of lncRNA NEAT1 by alveolar macrophages inhibited IL-6 secretion and NF-B nuclear transposition,suggesting that NEAT1 alleviated the inflammatory response of NR8383 alveolar macrophages through this mechanism.

关 键 词：脓毒症 肺泡巨噬细胞 lncRNA NEAT1 核因子-ΚB 

分 类 号：R563[医药卫生—呼吸系统]