MiR-127-5p通过靶向调节adipoR1促进骨关节炎软骨细胞的增殖

脂联素（adiponection）与骨关节炎（osteoarthritis, OA）的发病密切相关，且主要通过其受体adipoR1发挥作用。而骨关节炎中脂联素的表达是否受miRNA表达的影响却未见报道。本文旨在研究miR-127-5p对骨关节炎软骨细胞中脂联素及细胞增殖的影响。分离培养人原代OA软骨细胞及对应正常细胞，甲苯胺蓝染色和II型胶原免疫细胞化学染色进行鉴定。 Real-time PCR结果表明，OA软骨细胞中miR-127-5p的表达与正常软骨细胞中的相比较显著下降。MiR-127-5p转染可显著降低荧光素酶报告基因的荧光强度（P<0.05），表明adipoR1为miR-127-5p的靶向基因。MiR-127-5p mimic转染软骨细胞后，MTT法研究结果表明，miR-127-5p mimic 可显著促进软骨细胞增殖，Western 印迹结果表明，脂联素及其受体（adipoR1）表达显著上升，p65的表达以及p38、ERK1/2以及IkBα的磷酸化水平显著下降。ELISA结果表明，MMP-1、MMP-3、MMP-13的含量显著下降。实验结果提示，miR-127-5p通过靶向下调adipoR1及脂联素的表达，促进软骨细胞增殖，并且抑制NF-κB信号通路，进而抑制炎性反应。

MiR-127-5p Promotes Proliferation of Osteoarthritis Chondrocytes by Targeting AdipoR1

Adiponection is involved in the progression of osteoarthritis(OA) through receptor adipoR1. But whether the expression of adiponection is regulated by miRNA in the progression of OA remains unknown. This study was to analyze the effect of miR-127-5p on the expression of adiponection and the proliferation of OA chondrocytes. Chondrocytes were isolated from osteoarthritis patients and relative control patients. Toluidine blue staining and type II collagen immunocytochemistry staining assay were used to identify chondrocytes. Real-time PCR result showed that miR-127-5p was down-regulated in OA chondrocytes compared with normal chondrocytes. The dual luciferase reporter system showed that miR-127-5p could bind to the 3′UTR of adipoR1 and inhibit the expression of adipoR1. Furthermore, after OA chondrocytes were transfected with miR-127-5p mimic, the MTT result showed that the cell proliferation was significantly increased; Western blot showed that the expression of adipoR1 and adiponection were significantly increased. The expression of p65 and the phosphorylation level of p38,ERK1/2,IkBα were dramatically decreased. The ELISA showed that the concentration of MMP-1,MMP-3 and MMP-13 were dramatically decreased. These findings suggested that miR-127-5p promoted OA chondrocytes proliferation through downregulation of adipoR1 and adiponection, and inhibited inflammatory through inhibiting NF-κB signaling pathway. MiR-127-5p may have a protection effect on the progression of OA.