脂肪间充质干细胞外泌体miR-212-3p对成纤维样滑膜细胞的影响  

The Effect of fat mesenchymal stem cell exosome miR-212-3P on fibroblast synovial cells

作　　者：贾丙申[1] 于鹏[1] 焦拓 李君 李明 曲国欣 纪志华[1] 付昆[1] JIA Bingshen;YU Peng;JIAO Tuo;LI Jun;LI Ming;QU Guoxin;JI Zhihua;FU Kun(Department of Joint Trauma,the First Affiliated Hospital of Hainan Medical University,Haikou,Hainan,China,570102)

机构地区：[1]海南医学院第一附属医院关节创伤外科,海南海口570102

出　　处：《分子诊断与治疗杂志》2021年第10期1713-1717,1721,共6页Journal of Molecular Diagnostics and Therapy

基　　金：白求恩医学科学研究基金资助项目(N187CS)。

摘　　要：目的探讨脂肪间充质干细胞(ADSCs)外泌体miR-212-3p对类风湿性关节炎成纤维样滑膜细胞RA-FLS增殖、迁移和侵袭的影响及其作用机制。方法收集海南医学院第一附属医院类风湿性关节炎(RA)患者和健康者血清各20例,采用RT-qPCR检测血清中miR-212-3p的表达情况。体外细胞可分为4组:RA-FLS细胞单独培养(NC组)、RA-FLS细胞+Exo-ADSCs共培养(Exo组)、RA-FLS细胞+沉默外泌体miR-212-3p共培养(Exo-antagomiR-212-3p组)、敲降RA-FLS细胞SMAD1+沉默外泌体miR-212-3p共培养(si-SMAD1+Exo-antagomiR-212-3p组)。外源性调节ADSCs细胞中miR-212-3p和RA-FLS细胞中SMAD1的表达水平后,将ADSCs外泌体与RA-FLS细胞共培养,CCK-8和Transwell分别检测RA-FLS细胞增殖活力、迁移和侵袭能力;采用双荧光素酶报告基因验证miR-212-3p与SMAD1之间的靶向关系。结果与健康者相比,miR-212-3p在RA患者血清中低表达,差异有统计学意义(P<0.05);与NC组相比,Exo组中miR-212-3p表达显著上调,增殖活力、迁移和侵袭显著降低,差异无统计学意义(均P>0.05);miR-212-3p能够靶向结合SMAD1的3′UTR;同时与NC组相比,Exo组及si-SMAD1+Exo-antagomiR-212-3p组中SMAD1相对表达、RA-FLS细胞增殖活力、迁移和侵袭数显著降低,差异无统计学意义(均P<0.05),而Exo-antagomiR-212-3p组中SMAD1相对表达、RA-FLS细胞增殖活力、迁移和侵袭数与NC组比较,差异无统计学意义(均P>0.05)。结论ADSCs外泌体miR-212-3p在RA患者和RA-FLS细胞中低表达,其通过靶向下调SMAD1抑制RA-FLS细胞增殖、迁移和侵袭。Objective To investigate the effect of ADSCs exosome miR-212-3P on the proliferation,migration and invasion of RA-FLS cells and its mechanism.Methods Collect 20 serums of patients with rheumatoid arthritis(RA)and healthy people,and the expression of miR-212-3p in the serum was detected by RT-qPCR.In vitro cells can be divided into 4 groups:RA-FLS cells cultured alone(NC group),RA-FLS cells+Exo-ADSCs co-culture(Exo group),RA-FLS cells+silent exosomes miR-212-3p co-culture(Exo-antagomiR-212-3p group),knockdown RA-FLS cells SMAD1+silent exosomes miR-212-3p co-cultured(si-SMAD1+Exo-antagomiR-212-3p group).After exogenously regulating the expression level of miR-212-3p in ADSCs cells and SMAD1 in RA-FLS cells,ADSCs exosomes were co-cultured with RA-FLS cells,and the proliferation activity of RA-FLS cells was measured by CCK-8 and Transwell respectively,Migration and invasion ability;At the same time,dual luciferase reporter gene was used to verify the targeting relationship between miR-212-3p and SMAD1.Results Compared with healthy individuals,miR-212-3p wasunderexpressed in the serum of RA patients(P<0.05);compared with NC group,miR-212-3p expression was significantly up-regulated in Exo group,and proliferation activity,migration and invasion were significantly reduced(All P<0.05);miR-212-3p can target the 3′UTR of SMAD1;at the same time,compared with the NC group,the relative expression of SMAD1,in the Exo and si-SMAD1+Exo-antagomiR-212-3p groups RA-FLS cell proliferation activity,migration and invasion numbers were significantly reduced(all P<0.05),while the relative expression of SMAD1,RA-FLS cell proliferation activity,migration and invasion numbers in the Exo-antagomiR-212-3p group were not significantly different from those in the NC group(All P>0.05).Conclusion ADSCs exosome miR-212-3p was low expressed in RA patients and RA-FLS cells,and it inhibited RA-FLS cell proliferation,migration and invasion by targeting down regulation of SMAD1.

关 键 词：类风湿性关节炎 滑膜细胞 脂肪间充质干细胞移植 外泌体 miR-212-3p SMAD1 

分 类 号：R593.22[医药卫生—内科学]