miR-451对类风湿关节炎患者外周血单个核细胞增殖及炎症反应的影响

Effects of miR-451 on proliferation and inflammation of peripheral blood mononuclear cells from patients with rheumatoid arthritis

目的:探讨miR-451对类风湿关节炎(RA)患者外周血单个核细胞(PBMC)增殖及炎症反应的影响。方法:分离培养20例RA患者和5名健康人的PBMC。将RA患者的PBMC分为miR-NC组(脂质体法转染miR-451阴性对照)、miR-451组(转染miR-451 mimics)、miR-451+TNF-α组(转染miR-451mimics后用10μg/L的TNF-α处理24 h),以无处理细胞为对照组。qRT-PCR法检测细胞中miR-451的表达,MTT法检测细胞增殖抑制率,ELISA法检测细胞中IL-17、IL-1β、IL-6的表达,Western blot检测细胞中P65、P52、P50、IκBα蛋白的表达。结果:与健康人PBMC相比,RA患者PBMC增殖抑制率升高,miR-451、IL-17、IL-1β、IL-6的表达均上调(P<0.05)。与对照组和miR-NC组比较,miR-451组PBMC的增殖抑制率升高,IL-17、IL-1β、IL-6表达降低,P65、P52、P50蛋白表达下调,IκBα蛋白表达上调(P<0.05);与miR-451组比较,miR-451+TNF-α组PBMC增殖抑制率降低,细胞中IL-17、IL-1β、IL-6表达升高,P65、P52、P50蛋白表达上调,IκBα蛋白表达下调(P<0.05)。结论:miR-451可抑制RA患者PBMC的增殖和炎症因子表达,其机制与NF-κB信号通路失活有关。

Aim:To study the effects of miR-451 on proliferation and inflammatory response of peripheral blood mononuclear cells(PBMCs)from rheumatoid arthritis(RA)patients.Methods:PBMCs were isolated from the whole blood of 20 RA patients and 5 healthy people.The PBMCs from RA patients were allocated into 4 groups:the control group(without transfection or treatment),miR-NC group(transfected negative control miR-451),miR-451 group(transfected miR-451 mimics),miR-451+TNF-αgroup(transfected miR-451 mimics and then treated with 10μg/L TNF-α).The inhibition rate of proliferation was detected by MTT assay;ELISA assay was used to detect the expression of IL-17,IL-1βand IL-6 in PMBC;qRT-PCR was used to detect the expression of miR-451 in cells;Western blot was used to detect the protein expressions of P65,P52,P50 and IκBαin cells.Results:Compared with the PBMCs of healthy people,the proliferation inhibition rate,the expressions of miR-451,IL-17,IL-1βand IL-6 increased in PBMCs of RA patients(P<0.05).Compared with those of the control group and miR-NC group,the proliferation inhibition rate increased,and the expressions of IL-17,IL-1βand IL-6 decreased,the expressions of P65,P52 and P50 proteins were down-regulated,and the expression of IκBαprotein was up-regulated in PBMCs of miR-451 group(P<0.05).Compared with those of the miR-451 group,the proliferation inhibition rate decreased,the expressions of IL-17,IL-1βand IL-6 increased,the expressions of P65,P52 and P50 proteins were up-regulated,and the expression of IκBαprotein was down-regulated in PBMCs of miR-451+TNF-αgroup(P<0.05).Conclusion:miR-451 could inhibit the proliferation and inflammatory reaction of PBMCs of RA patients through inactivating NF-κB signaling pathway.