软骨细胞来源的过表达 miR-214-3p 的外泌体促进 M2 巨噬细胞极化和血管生成，从而缓解Legg-Calvé-Perthes 病。
Exosomes from chondrocytes overexpressing miR-214-3p facilitate M2 macrophage polarization and angiogenesis to relieve Legg Calvé-Perthes disease.
机构信息
Department of Orthopaedics, the First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, PR China.
Department of Orthopaedics, the First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, PR China.
出版信息
Cytokine. 2023 Aug;168:156233. doi: 10.1016/j.cyto.2023.156233. Epub 2023 May 27.
OBJECTIVE
Legg-Calvé-Perthes disease (LCPD) is a partial or total necrosis of femoral head bone caused by blood supply disorder and its etiology is not clear. Studies have revealed that microRNA-214-3p (miR-214-3p) plays a vital role in LCPD, however, its exact mechanism is still unclear. In this study, we investigated the potential role of chondrocytes-derived exosomes carrying miR-214-3p (exos-miR-214-3p) in the pathogenesis of LCPD.
METHODS
RT-qPCR was performed to evaluate miR-214-3p expression level in femoral head cartilage, serum and chondrocytes of patients with LCPD, as well as dexamethasone (DEX)-exposed TC28 cells. Effects of exos-miR-214-3p on the proliferation and apoptosis were verified via MTT assay, TUNEL staining and caspase3 activity assay. The M2 macrophage markers were assessed by flow cytometry, RT-qPCR and Western blot. Moreover, angiogenic effects of human umbilical vein endothelial cells (HUVECs) were tested using CCK-8 and tube formation assays. Bioinformatics prediction, luciferase assay and ChIP were applied to verify the association between ATF7, RUNX1 and miR-214-3p.
RESULTS
miR-214-3p was found to be decreased in patients with LCPD and DEX-treated TC28 cells, of which overexpression promoted cell proliferation and suppressed apoptosis. Mechanistically, exos-miR-214-3p facilitated M2 polarization by ATF7/TLR4 axis and HUVECs angiogenesis via RUNX1/VEGFA axis.
CONCLUSION
miR-214-3p alleviates LCPD by promoting M2 polarization of macrophages and angiogenesis.
目的
莱格-卡尔弗-佩尔特斯病（LCPD）是一种由血液供应紊乱引起的股骨头部分或全部坏死，其病因尚不清楚。研究表明，微小 RNA-214-3p（miR-214-3p）在 LCPD 中发挥着重要作用，但其确切机制尚不清楚。在本研究中，我们研究了携带 miR-214-3p（exos-miR-214-3p）的软骨细胞衍生外泌体在 LCPD 发病机制中的潜在作用。
方法
通过 RT-qPCR 评估 LCPD 患者股骨头软骨、血清和软骨细胞以及地塞米松（DEX）暴露的 TC28 细胞中 miR-214-3p 的表达水平。通过 MTT 测定、TUNEL 染色和 caspase3 活性测定验证 exos-miR-214-3p 对增殖和凋亡的影响。通过流式细胞术、RT-qPCR 和 Western blot 评估 M2 巨噬细胞标志物。此外，通过 CCK-8 和管形成测定检测人脐静脉内皮细胞（HUVECs）的血管生成作用。应用生物信息学预测、荧光素酶测定和 ChIP 验证 ATF7、RUNX1 和 miR-214-3p 之间的关联。
结果
miR-214-3p 在 LCPD 患者和 DEX 处理的 TC28 细胞中表达降低，其中过表达促进细胞增殖并抑制凋亡。机制上，exos-miR-214-3p 通过 ATF7/TLR4 轴促进 M2 极化，并通过 RUNX1/VEGFA 轴促进 HUVECs 血管生成。
结论
miR-214-3p 通过促进巨噬细胞 M2 极化和血管生成来缓解 LCPD。
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