微小RNA-147负向调节大鼠巨噬细胞中Toll样受体7的表达，并减轻 pristane 诱导的大鼠类风湿性关节炎。
MicroRNA-147 negatively regulates expression of toll-like receptor-7 in rat macrophages and attenuates pristane induced rheumatoid arthritis in rats.
作者信息
Zhao Wei, Li Dai, Su Yuqiang, Zhao Haikang, Pang Weiwei, Sun Yang, Wu Shengjun
机构信息
Department of Rehabilitation Medicine, The Second Affiliated Hospital of Xi'an Medical University Xi'an 710038, Shaanxi, China.
Department of Anesthesiology, Chang Hai Hospital, Naval Military Medical University Shanghai 200433, China.
出版信息
Am J Transl Res. 2019 Apr 15;11(4):2219-2231. eCollection 2019.
UNLABELLED
Background/Introduction: Aberrant expression of Toll like receptors (TLR) plays a vital role in pathogenesis of rheumatoid arthritis (RA). Micro RNAs (miRs) could play important role in the related signaling pathways. The present study was undertaken to establish the link between miR-147 and TLR-7 in rat macrophages () and in pristane (PS) induced arthritic rats.
METHODOLOGY
Dual luciferase assay was done to confirm the interaction between miR-147 and TLR-7. The effect of miR-147 on regulation of TLR-7 was done by RT-qPCR and Immunoblotting studies in rat macrophages (ATCC CRL-2192) after treating them with miR-147 mimics and inhibitors. R-848 (Imiquimod) was used as TLR-7 stimulant, the mRNA and protein expression levels of IFN-β and TNF-α were recorded to determine the regulation of TLR-7. The levels of miR-147 and TLR-7 were evaluated during induction of rat bone marrow derived macrophage in the PS induced rat macrophages and spleens of methotrexate exposed rats. The miR-147 mimics was injected intraperitoneal to the PS treated rats and the severity of arthritis was studied.
RESULTS
The study confirmed TLR-7 mRNA as the potential target of miR-147 in rats. Alterations in miR-147 by transfecting mimics or inhibitors in ATCC CRL-2192 cells exhibited suppression and amelioration of TLR-7 and cytokine expression. The alteration in expression of miR-147 was inversely correlated with expression of TLR-7 during bone marrow derived macrophages induction in PS exposed cells and spleens. The abnormal expression was reversed in spleens of methotrexate treated arthritic rats. The treatment of miR-147 mimic caused suppression in expression of TLR-7 and improved the severity of arthritis in PS induced arthritic rats.
CONCLUSIONS
MiR-147 inversely regulates the TLR-7 signaling by targeting TLR-7 itself both and . The study provides a novel approach for conditions involving abnormal TLR-7 expression in arthritis.
未标记
背景/引言：Toll样受体（TLR）的异常表达在类风湿关节炎（RA）的发病机制中起着至关重要的作用。微小RNA（miR）可能在相关信号通路中发挥重要作用。本研究旨在建立大鼠巨噬细胞（）和 pristane（PS）诱导的关节炎大鼠中 miR-147 与 TLR-7 之间的联系。
方法
采用双荧光素酶报告基因检测法来确认 miR-147 与 TLR-7 之间的相互作用。在用 miR-147 模拟物和抑制剂处理大鼠巨噬细胞（ATCC CRL-2192）后，通过 RT-qPCR 和免疫印迹研究来观察 miR-147 对 TLR-7 调节的影响。使用 R-848（咪喹莫特）作为 TLR-7 刺激剂，记录 IFN-β 和 TNF-α 的 mRNA 和蛋白表达水平以确定 TLR-7 的调节情况。在 PS 诱导的大鼠巨噬细胞和甲氨蝶呤暴露大鼠的脾脏中诱导大鼠骨髓来源巨噬细胞的过程中，评估 miR-147 和 TLR-7 的水平。将 miR-147 模拟物腹腔注射到 PS 处理的大鼠中，并研究关节炎的严重程度。
结果
该研究证实 TLR-7 mRNA 是大鼠中 miR-147 的潜在靶点。在 ATCC CRL-2192 细胞中转染模拟物或抑制剂改变 miR-147 后，显示出 TLR-7 和细胞因子表达的抑制和改善。在 PS 暴露细胞和脾脏中骨髓来源巨噬细胞诱导过程中，miR-147 表达的改变与 TLR-7 的表达呈负相关。在甲氨蝶呤治疗的关节炎大鼠脾脏中，异常表达得到逆转。miR-147 模拟物的治疗导致 TLR-7 表达受到抑制，并改善了 PS 诱导的关节炎大鼠的关节炎严重程度。
结论
miR-147 通过靶向 TLR-7 本身在 和 中反向调节 TLR-7 信号通路。该研究为涉及关节炎中 TLR-7 异常表达的情况提供了一种新方法。
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