微小 RNA-146a-5p 通过抑制 SDF-1/CXCR4 诱导的软骨细胞自噬来缓解骨关节炎的发病机制。
MicroRNA-146a-5p alleviates the pathogenesis of osteoarthritis by inhibiting SDF-1/CXCR4-induced chondrocyte autophagy.
机构信息
Department of Sports Medicine, The First Affiliated Hospital, Kunming Medical University, Kunming 650032, Yunnan, China.
Department of Sports Medicine, The First Affiliated Hospital, Kunming Medical University, Kunming 650032, Yunnan, China.
出版信息
Int Immunopharmacol. 2023 Apr;117:109938. doi: 10.1016/j.intimp.2023.109938. Epub 2023 Feb 28.
BACKGROUND
SDF-1/CXCR4 signaling promotes osteoarthritis (OA) development. CXCR4 is a potential target of miR-146a-5p. This study investigated the therapeutic role and the underlying mechanism of miR-146a-5p in OA.
METHODS
Human primary chondrocytes C28/I2 were stimulated with SDF-1. Cell viability and LDH release were examined. Chondrocyte autophagy was assessed using Western blot analysis, ptfLC3 transfection, and transmission electron microscopy. MiR-146a-5p mimics were transfected into C28/I2 cells to investigate the role of miR-146a-5p in SDF-1/CXCR4-induced autophagy of chondrocytes. An SDF-1-induced rabbit OA model was established to investigate the therapeutic role of miR-146a-5p in OA. Histological staining was performed to observe the morphology of osteochondral tissue.
RESULTS
SDF-1/CXCR4 signaling promoted autophagy in C28/I2 cells, as demonstrated by increased LC3-II protein expression and autophagic flux induced by SDF-1. SDF-1 treatment significantly inhibited cell proliferation while promoting necrosis and autophagosome formation in C28/I2 cells. In the presence of SDF-1, miR-146a-5p overexpression in C28/I2 cells suppressed CXCR4 mRNA expression, LC3-II and Beclin-1 protein expression, LDH release, and autophagic flux. In addition, SDF-1 increased the autophagy of chondrocytes in rabbits and promoted the development of OA. Compared with the negative control, miR-146a-5p significantly reduced the morphological abnormalities of the rabbit cartilage that were induced by SDF-1, as well as the number of LC3-II-positive cells, protein expression of LC3-II and Beclin 1, and mRNA expression of CXCR4 in osteochondral tissue. These effects were reversed by the autophagy agonist rapamycin.
CONCLUSIONS
SDF-1/CXCR4 promotes OA development by enhancing chondrocyte autophagy. MicroRNA-146a-5p may alleviate OA by suppressing CXCR4 mRNA expression and SDF-1/CXCR4-induced chondrocyte autophagy.
背景
SDF-1/CXCR4 信号通路促进骨关节炎（OA）的发展。CXCR4 是 miR-146a-5p 的一个潜在靶点。本研究探讨了 miR-146a-5p 在 OA 中的治疗作用及其潜在机制。
方法
用 SDF-1 刺激人原代软骨细胞 C28/I2，检测细胞活力和 LDH 释放。用 Western blot 分析、ptfLC3 转染和透射电镜评估软骨细胞自噬。转染 miR-146a-5p 模拟物到 C28/I2 细胞中，以研究 miR-146a-5p 在 SDF-1/CXCR4 诱导的软骨细胞自噬中的作用。建立 SDF-1 诱导的兔 OA 模型，以研究 miR-146a-5p 在 OA 中的治疗作用。进行组织学染色以观察骨软骨组织的形态。
结果
SDF-1/CXCR4 信号促进 C28/I2 细胞自噬，表现为 SDF-1 诱导的 LC3-II 蛋白表达增加和自噬流。SDF-1 处理显著抑制细胞增殖，同时促进 C28/I2 细胞坏死和自噬体形成。在 SDF-1 存在的情况下，C28/I2 细胞中 miR-146a-5p 的过表达抑制了 CXCR4 mRNA 表达、LC3-II 和 Beclin-1 蛋白表达、LDH 释放和自噬流。此外，SDF-1 增加了兔软骨细胞的自噬，促进了 OA 的发展。与阴性对照组相比，miR-146a-5p 显著减少了 SDF-1 诱导的兔软骨的形态异常，以及软骨组织中 LC3-II 阳性细胞数、LC3-II 和 Beclin 1 蛋白表达以及 CXCR4 mRNA 表达。这些作用可被自噬激动剂 rapamycin 逆转。
结论
SDF-1/CXCR4 通过增强软骨细胞自噬促进 OA 的发展。miR-146a-5p 可能通过抑制 CXCR4 mRNA 表达和 SDF-1/CXCR4 诱导的软骨细胞自噬来减轻 OA。
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