miR-155通过SOCS1/STAT3途径调控类风湿性关节炎中炎症反应和Th17/Treg失衡OA北大核心CSTPCD
MiR-155 regulates inflammatory responses and Th17/Treg imbalances in rheumatoid arthritis through the SOCS1/STAT3 pathway
目的 在类风湿性关节炎(RA)中探究miR-155和细胞因子信号转导抑制因子1(SOCS1)的表达及作用机制.方法 采用RT-PCR和流式细胞术检测miR-155和Th17、Treg细胞在RA患者(RA组)和对照组(HC组)外周血中的表达差异;生物信息学和双荧光素酶基因报告实验检测miR-155和SOSC1的调控关系;分离RA患者外周CD4+T细胞,将沉默miR-155与SOCS1表达的miR-155 inhibitor和si-SOCS1及各自的阴性对照序列分别或联合转染入CD4+T细胞中,并将细胞分为:miR-NC组、miR-155 inhibitor组、miR-155 inhibitor+si-NC组和miR-155 inhibitor+si-SOCS1组.使用Th17诱导分化液处理上述细胞后,采用流式细胞术检测各组CD4+T细胞中Th17比率,Western blot实验检测细胞中p-STAT3/STAT3比值.结果 与HC组相比,RA患者中miR-155、Th17比率升高(P<0.01),Treg细胞比率降低(P<0.01);miR-155可靶向抑制SOCS1表达.与miR-NC组相比,miR-155 inhibitor组、miR-155 inhibitor+si-NC组和miR-155 inhibitor+si-SOCS1组中Th17 比率、p-STAT3/STAT3 比值均降低(P<0.01);与miR-155 inhibitor组相比,miR-155 inhibitor+si-SOCS1组CD4+T细胞中Th17比率、p-STAT3/STAT3比值均升高(P<0.05).结论 在RA患者中表达升高的miR-155可能通过SOCS1/STAT3途径来介导CD4+T细胞的Th17分化,从而参与RA患者外周Th17/Treg细胞失衡.
Objective This research aimed to investigate the expression and mechanism of miR-155 and suppressor of cytokine signaling 1(SOCS1)in rheumatoid arthritis(RA).Methods RT-PCR and flow cytometry were applied to detect the expression differences of miR-155,Th17,and Treg cells in peripheral blood of RA patients(RA group)and control group(HC group).Bioinformatics analysis and dual-luciferase reporter assay were conducted to investigate the regulatory relationship between miR-155 and SOCS1.CD4+T cells were isolated from peripheral blood of RA patients and transfected with miR-155 inhibitor,si-SOCS1,and their respective negative control sequences,and divided into four groups:miR-NC group,miR-155 inhibitor group,miR-155 inhibitor+si-NC group,and miR-155 inhibitor+si-SOCS1 group.The cells were treated with Th17-inducing differentiation medium,and flow cytometry was used to determine the ratio of Th17 cells in each group of CD4+T cells.Western blot was used to determine the ratio of p-STAT3/STAT3 in the cells.Results Compared to the HC group,RA patients showed increased expression of miR-155 and Th17 ratio(P<0.01),and decreased Treg cell ratio(P<0.01).MiR-155 could target and inhibit the expression of SOCS1.Compared to the miR-NC group,the miR-155 inhibitor group,miR-155 inhibitor+si-NC group,and miR-155 inhibitor+si-SOCS1 group showed decreased Th17 ratio and p-STAT3/STAT3 ratio(P<0.01).Compared to the miR-155 inhibitor group,the miR-155 inhibitor+si-SOCS1 group exhibited increased Th17 ratio and p-STAT3/STAT3 ratio in CD4+T cells(P<0.05).Conclusion Elevated expression of miR-155 in RA patients may mediate the differentiation of CD4+T cells into Th17 cells through the SOCS1/STAT3 pathway,contributing to the imbalance of Th17/Treg cells in peripheral blood of RA patients.
张玉红;单新洁;周俊
新疆生产建设兵团医院肾病内科(乌鲁木齐 830000)新疆维吾尔自治区儿童医院检验科(乌鲁木齐 830010)
临床医学
类风湿性关节炎miR-155细胞因子信号转导抑制因子1Th17/Treg失衡
rheumatoid arthritismiR-155suppress of cytokine signaling 1(SOCS1)Th17/Treg imbalance
《实用医学杂志》 2024 (013)
1791-1796 / 6
兵团科技攻关与成果转化计划项目(编号:2020AD008)
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