LncRNA MALAT1通过调控miR-181d-5p对脂多糖诱导的库普弗细胞增殖活性和炎性因子表达的影响  

Effect of LncRNA MALAT1 on Lipopolysaccharide-induced Kupffer Cell Proliferation and Inflammatory Factor Expression via Regulation of miR-181d-5p

作　　者：杜娜 段少琼 宋波 李开林 颜悦蓉 刘悦 DU Na;DUAN Shaoqiong;SONG Bo;LI Kailin;YAN Yuerong;LIU Yue(Department of Infectious Diseases,Suizhou Central Hospital,Suizhou Hospital Affiliated to Hubei University of Medicine,Suizhou,Hubei,441300,China;Department of Gastroenterology,Xiangyang Central Hospital,Affiliated Hospital of Hubei University of Arts and Science,Xiangyang,Hubei,441021,China)

机构地区：[1]湖北医药学院附属随州医院随州市中心医院感染科,湖北省随州市441300 [2]湖北文理学院附属医院襄阳市中心医院消化内科,湖北省襄阳市441021

出　　处：《医学分子生物学杂志》2023年第5期426-431,共6页Journal of Medical Molecular Biology

基　　金：湖北省卫生和计划生育委员会基金(No.WJ2017H039)。

摘　　要：目的探讨长链非编码RNA肺腺癌转移相关转录本1(LncRNA MALAT1)对脂多糖(LPS)诱导的库普弗细胞(kupffer cells,KCs)存活和炎性因子表达的影响及其可能作用机制。方法采用LPS诱导KCs建立细胞损伤模型,随机分组为对照(Con)组、LPS组、LPS+si-NC组、LPS+si-MALAT1组、LPS+miR-NC组、LPS+miR-181d-5p组、LPS+si-MALAT1+anti-miR-NC组、LPS+si-MALAT1+anti-miR-181d-5p组;实时荧光定量聚合酶链式反应(qRT-PCR)检测LncRNA MALAT1、miR-181d-5p的表达量;细胞计数试剂盒8(CCK-8)法检测细胞增殖活性;酶联免疫吸附实验(ELISA)检测白细胞介素(IL)-6、IL-12、肿瘤坏死因子α(TNF-α)的水平;双荧光素酶报告实验检测LncRNA MALAT1与miR-181d-5p的靶向关系。结果与Con组比较,LPS组LncRNA MALAT1的表达量升高(P<0.05),IL-6、IL-12、TNF-α的水平升高(P<0.05),miR-181d-5p的表达量降低(P<0.05),细胞增殖活性降低(P<0.05)。与LPS+si-NC组比较,LPS+si-MALAT1组细胞增殖活性升高(P<0.05),IL-6、IL-12、TNF-α的水平降低(P<0.05)。LncRNA MALAT1可负向调控miR-181d-5p的表达;与LPS+miR-NC组比较,LPS+miR-181d-5p组细胞增殖活性升高(P<0.05),IL-6、IL-12、TNF-α的水平降低(P<0.05)。与LPS+si-MALAT1+antimiR-NC组比较,LPS+si-MALAT1+anti-miR-181d-5p组细胞增殖活性降低(P<0.05),IL-6、IL-12、TNF-α的水平升高(P<0.05)。结论干扰LncRNA MALAT1表达可通过促进miR-181d-5p表达而促进LPS诱导的KCs存活及抑制细胞炎性因子表达。Objective To investigate the effect of LncRNA MALAT1 on lipopolysaccharide(LPS)-induced survival and expression of inflammatory factors in kupffer cells(KCs)and its possible mechanism.Methods LPS-induced KCs were used to establish a cell injury model and were randomly divided into 8 groups:Con group,LPS group,LPS+si-NC group,LPS+si-MALAT1 group,LPS+miR-NC group,LPS+miR-181d-5p group,LPS+si-MALAT1+anti-miR-NC group,LPS+si-MALAT1+anti-miR-181d-5p group.The expression levels of LncRNA MALAT1 and miR-181d-5p were detected by qRT-PCR.Cell viability was detected by CCK-8 method.The levels of IL-6,IL-12 and TNF-αwere detected by ELISA.The targeting relationship between LncRNA MALAT1 and miR-181d-5p was detected by dual luciferase gene reporter assay.Results The expression of LncRNA MALAT1 in the LPS group was increased(P<0.05),the levels of IL-6,IL-12 and TNF-αwere increased(P<0.05),while the expression of miR-181d-5p was decreased(P<0.05),and the cell viability was decreased(P<0.05),when compared with those in the Con group.The cell survival rate in the LPS+si-MALAT1 group was increased(P<0.05),while the levels of IL-6,IL-12 and TNF-αwere decreased(P<0.05),when compared with those in the LPS+si-NC group.The LncRNA MALAT1 negatively regulated the expression of miR-181d-5p.The cell survival rate in the LPS+miR-181d-5p group was increased(P<0.05),while the levels of IL-6,IL-12 and TNF-αwere decreased(P<0.05),when compared with those in the LPS+miRNC group.The survival rate in the LPS+si-MALAT1+anti-miR-181d-5p group was decreased(P<0.05),while the levels of IL-6,IL-12,TNF-αwere increased(P<0.05),when compared with those in the LPS+si-MALAT1+anti-miR-NC group.Conclusion Interfering with the expression of LncRNA MALAT1 could promote the survival of LPS-induced KCs and inhibit the expression of inflammatory factors by promoting the expression of miR-181d-5p.

关 键 词：LncRNA MALAT1 miR-181d-5p 脂多糖 库普弗细胞 炎症因子 

分 类 号：R363[医药卫生—病理学]