RA患者microRNA-155调节免疫细胞分化的机制研究  

Mechanism of microRNA-155 regulating immune cell differentiation in patients with rheumatoid arthritis

作　　者：杨瑞宾 国学红 周成英 吴雪梅 Yang Ruibin;Guo Xuehong;Zhou Chengying;Wu Xuemei(Xingyi City People’s Hospital,Xingyi 562400,Guizhou,China.)

机构地区：[1]兴义市人民医院,贵州兴义562400

出　　处：《贵州医药》2023年第5期675-677,687,F0003,共5页Guizhou Medical Journal

基　　金：贵州省卫健委科技计划项目(编号:gzwjkj2019-1-046)。

摘　　要：目的探讨类风湿性关节炎(RA)患者miR-155如何调节CD4+T细胞亚群Th17及Treg细胞的分化及其特异性转录因子和细胞因子的表达,了解miR-155主要作用的靶基因及其调节的JAK-SATA信号通路在RA中的致病机制。方法采集62例RA患者(包括37例轻度类风湿关节炎患者与25例重度类风湿关节炎患者)及30例健康对照组外周血提取RNA,采用实时荧光定量逆转录聚合酶链反应(RT-qPCR)的方法检测各组研究对象外周血miR-155、SOCS1、JAK3、RORγt、SATA3、SATA5、Foxp3等基因的mRNA表达情况;采用免疫印迹分析(Western boltting)检测SOCS1、JAK3、RORγt、SATA3/5、Foxp3靶基因及相关信号通路中重要蛋白的表达情况;采用免疫荧光法分别检测各组研究对象血清中IL-2、IL-6、IL-17、IL-10等细胞因子的表达情况。采用电化学发光法、免疫散射比浊法及胶乳增强免疫比浊法分别检测各组研究对象血清中A-CCP、CRP、ASO与RF含量。结果RA患者外周血中miR-155 mRNA、JAK3mRNA、SATA3mRNA、RORγt mRNA及其蛋白表达水平明显高于健康对照组(P<0.05),且与疾病严重程度相关;而SOCS1mRNA、SATA5mRNA、Foxp3 mRNA及其蛋白表达水平明显低于健康对照组,且与疾病严重程度相关(P<0.05);RA患者IL-2、IL-6、IL-10、IL-17水平明显高于健康对照组(P<0.05)且与疾病严重程度相关;RA患者血清中A-CCP、RF含量显著高于健康对照组(P<0.05),且与疾病严重程度相关。结论RA患者外周血miR-155表达明显升高,在CD4+T细胞分化过程中,miR-155可能通过抑制SOCS1的表达而上调JAK/STAT信号通路中SATA3/RORrt通路分子以促进Th17细胞的分化及其细胞因子IL-6、IL-17的表达;可能通过下调JAK/STAT信号通路中SATA5/Foxp3通路分子以抑制Treg细胞的分化及分泌。Objective To clarify how miR-155 regulates CD4+T cell subsets differentiation that including Th17 and Treg cells and the expression of specific transcription factors and cytokines in rheumatoid arthritis(RA)patients,and clarify the target genes of miR-155 and the pathogenic mechanism of JAK-SATA signal pathway in RA.Methods RNA was extracted from peripheral blood of 62 patients with RA that including 37 patients with mild rheumatoid arthritis and 25 patients with severe rheumatoid arthritis,and 30 healthy controls.The mRNA levels of miR-155,SOCS1,JAK3,RORγt,SATA3,SATA5,and Foxp3 in peripheral blood of subjects in each group were detected by RT-qPCR.The protein expression of SOCS1,JAK3,RORγt,SATA3,SATA5,and Foxp3 in the signaling pathways was detected by western boltting.The expression of cytokines IL-2,IL-6,IL-17 and IL-10 in the serum of the subjects in each group was detected by Immunofluorescence method.A-CCP,RF,ASO,CRP using to Electrochemical luminescence,Immunoturbidimetric assay and so on.Results The mRNA levels of miR-155,JAK3,SATA3,RORγt and its protein in peripheral blood of patients with RA were significantly higher than the healthy control group and there were statistical significantly differences(P<0.05),and were related to the severity of the disease.The mRNA levels of SOCS1,SATA5,Foxp3 and its protein in peripheral blood of patients with RA were significantly lower than the healthy control group,and were correlated with the severity of the disease(P<0.05).The levels of IL-2,IL-6,IL-10 and IL-17 in RA patients were significantly higher than the healthy control(P<0.05),and were related to the severity of the disease.The levels of A-CCP,RF in RA patients were significantly higher than that healthy controls(P<0.05),and were related to the severity of the disease.Conclusion The expression of miR-155 in peripheral blood of RA patients is increased significantly.During the differentiation of CD4+T cells,miR-155 up regulated SATA3/RORrt pathway in JAK/STAT signal pathway by inhibiting the expression of

关 键 词：MIR-155 JAK-SATA信号通路 SATA3 RORΓT SATA5 FOXP3 

分 类 号：R593.22[医药卫生—内科学]