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微小RNA-19通过靶向类风湿关节炎中的Toll样受体2抑制成纤维样滑膜细胞细胞因子释放。

MiR-19 suppresses fibroblast-like synoviocytes cytokine release by targeting toll like receptor 2 in rheumatoid arthritis.

作者信息

Li Zongyu, Cai Jinfang, Cao Xuecheng

机构信息

Department of Traumatic Orthopedic Surgery, The General Hospital of Jinan Military Command Jinan 250031, Shandong, China.

出版信息

Am J Transl Res. 2016 Dec 15;8(12):5512-5518. eCollection 2016.

Abstract

Fibroblast-like synoviocytes (FLS) play an important role in the pathogenesis of rheumatoid arthritis (RA) through participating in joint tissue inflammation and joint damage. MicroRNAs are a kind of small non-coding RNAs that can regulate gene expression in the transcription level to affect cell behaviors. This study intended to investigate the expression of miR-19 in FLS from RA patients and related mechanism. A total of 126 RA patients were selected in this study. MiR-19 expression in FLS was detected by qRT-PCR. Toll like receptor 2 (TLR2) protein expression was tested by Western blot. MiR-19 target genes were confirmed by bioinformatics analysis and luciferase reporter assay. The impact of miR-19 on the expression of TLR2, interleukin 6 (IL-6), and matrix metalloproteinase 3 (MMP-3) in FLS were analyzed by cell transfection and Western blot. MiR-19 expression in FLS from RA patients was significantly downregulated compared with control (P < 0.05), while TLR2 level was increased (P < 0.05). Bioinformatics analysis and luciferase reporter assay confirmed that TLR2 was the target gene of miR-19. Transfection of miR-19 mimic or miR-19 inhibitor obviously suppressed or increased TLR2 expression, and reduced or promoted release of cytokines IL-6 and MMP-3 in FLS, respectively. In conclusion, MiR-19 expression was downregulated in FLS from RA patients, leading to increased TLR2 expression and enhanced cytokines release.

摘要

成纤维样滑膜细胞(FLS)通过参与关节组织炎症和关节损伤在类风湿关节炎(RA)的发病机制中起重要作用。微小RNA是一类小的非编码RNA,其可在转录水平调节基因表达以影响细胞行为。本研究旨在探讨RA患者FLS中miR-19的表达及相关机制。本研究共纳入126例RA患者。采用qRT-PCR检测FLS中miR-19的表达。通过蛋白质印迹法检测Toll样受体2(TLR2)蛋白表达。通过生物信息学分析和荧光素酶报告基因测定法确认miR-19的靶基因。通过细胞转染和蛋白质印迹法分析miR-19对FLS中TLR2、白细胞介素6(IL-6)和基质金属蛋白酶3(MMP-3)表达的影响。与对照组相比,RA患者FLS中miR-19的表达显著下调(P<0.05),而TLR2水平升高(P<0.05)。生物信息学分析和荧光素酶报告基因测定法证实TLR2是miR-19的靶基因。转染miR-19模拟物或miR-19抑制剂分别明显抑制或增加FLS中TLR2的表达,并降低或促进细胞因子IL-6和MMP-3的释放。总之,RA患者FLS中miR-19表达下调,导致TLR2表达增加和细胞因子释放增强。

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