LncRNA MIR22HG通过海绵吸附miR-22-5p对类风湿关节炎成纤维样滑膜细胞增殖、凋亡和炎性反应的影响  

Effects of lncRNA MIR22HG on proliferation,apoptosis and inflammatory response of rheumatoid arthritis fibroblast-like synoviocytes by sponge adsorption of miR-22-5p

作　　者：杨舟[1] 林书典[1] 詹宇威 肖璐 符克英[1] 黄小蝶[1] Yang Zhou;Lin Shudian;Zhan Yuwei;Xiao Lu;Fu keying;Huang Xiaodie(Dept of Rheumatology,Affiliated Hainan Hospital of Hainan Medical University,Hainan General Hospital Hainan General Hospital,Haikou 570311)

机构地区：[1]海南省人民医院,海南医学院附属海南医院风湿免疫科,海口570311

出　　处：《安徽医科大学学报》2023年第3期405-412,共8页Acta Universitatis Medicinalis Anhui

基　　金：海南省自然科学基金面上项目(编号:820MS128);海南省省级临床医学中心建设项目[编号:琼卫医涵(2021)75号]。

摘　　要：目的探讨长链非编码RNA(LncRNA)MIR22HG对类风湿关节炎(RA)成纤维样滑膜细胞(FLSs)增殖、凋亡和炎性反应的影响及其分子机制。方法收集在本院接受治疗的37例RA患者及30例关节创伤患者的滑膜组织样本,采用qRT-PCR检测滑膜组织中MIR22HG和miR-22-5p表达水平。体外分离、培养和鉴定人RA-FLSs,将MIR22HG siRNA干扰质粒(si-MIR22HG)及其阴性对照质粒(si-NC)和miR-22-5p inhibitor及其阴性对照(inhibitor-NC)分别或同时转染至RA-FLSs中,qRT-PCR检测细胞中MIR22HG和miR-22-5p表达水平;CCK-8检测各组细胞增殖活性;Annexin Ⅴ-FITC/PI检测各组细胞凋亡率;ELISA检测各组细胞上清液中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β和IL-6水平;Western blot检测各组细胞中Bcl-2、Bax和Cleaved caspase-3蛋白表达水平。双荧光素酶报告基因实验验证MIR22HG与miR-22-5p之间的靶向关系。结果与关节创伤患者比较,RA患者滑膜组织中MIR22HG表达水平升高(P<0.05),而miR-22-5p表达水平降低(P<0.05)。干扰MIR22HG可抑制RA-FLSs增殖活性,降低细胞上清液中TNF-α、IL-1β和IL-6水平及细胞中Bcl-2蛋白表达水平(P<0.05),提高细胞凋亡率、miR-22-5p及Bax、Cleaved casepase-3等蛋白表达水平(P<0.05)。然而,抑制miR-22-5p表达可逆转MIR22HG基因沉默对RA-FLSs增殖、凋亡和炎症的影响(P<0.05)。双荧光素酶报告实验显示,miR-22-5p是MIR22HG潜在的下游靶miRNA。结论MIR22HG在RA患者关节滑膜组织中高表达,其可能通过靶向下调miR-22-5p表达促进RA-FLSs增殖及炎性反应,并抑制细胞凋亡。Objective To explore the effects of long non-coding RNA(LncRNA)MIR22HG on proliferation,apoptosis and inflammatory response of rheumatoid arthritis(RA)fibroblast-like synoviocytes(FLSs)and its molecular mechanism.Methods Synovial tissue samples were collected from 37 RA patients and 30 joint trauma patients in our hospital,and the expression levels of MIR22HG and miR-22-5p in synovial tissue were detected by qRT-PCR.RA-FLSs in human was isolated,cultured and identified in vitro.MIR22HG siRNA interference plasmid(si-MIR22HG)and its negative control plasmid(si-NC),miR-22-5p inhibitor and its negative control(inhibitor-NC)were transfected into RA-FLSs respectively or simultaneously.The expression levels of MIR22HG and miR-22-5p were detected by qRT-PCR.CCK-8 was used to detect the proliferation activity of cells in various groups.AnnexinⅤ-FITC/PI was used to detect the apoptosis rates of cells in various groups.ELISA was used to detect the levels of TNF-α,IL-1β and IL-6 in the supernatant of cells in various groups.Western blot was used to detect the protein expression levels of Bcl-2,Bax and Cleaved caspase-3 of cells in various groups.The targeting relationship between MIR22HG and miR-22-5p was verified by dual luciferase reporter gene assay.Results Compared with joint trauma patients,the expression level of MIR22HG in synovial tissues of RA patients increased(P<0.05),while the expression level of miR-22-5p decreased(P<0.05).Interference with MIR22HG inhibited the proliferation activity of RA-FLSs,decreased the levels of TNF-α,IL-1β and IL-6 in cell supernatant and the protein expression level of Bcl-2 in cells(P<0.05),and increased the apoptosis rate,the expression level of miR-22-5p and the protein expression levels of Bax and Cleaved casepase-3(P<0.05).However,inhibition of miR-22-5p expression reversed the effects of MIR22HG gene silencing on proliferation,apoptosis and inflammation of RA-FLSs(P<0.05).Dual luciferase reporting assay showed that miR-22-5p was a potential downstream miRNA target of MIR22HG

关 键 词：类风湿关节炎 成纤维样滑膜细胞 LncRNA MIR22HG miR-22-5p 细胞增殖 细胞凋亡 炎性反应 

分 类 号：R593.22[医药卫生—内科学]