Department of Stomatology, Shenzhen Hospital, Southern Medical University, Shenzhen, 518033, China.
Department of Orthodontics, Beijing Stomatological Hospital, Capital Medical University, Beijing, 100006, China.
Arch Oral Biol. 2021 Apr;124:105060. doi: 10.1016/j.archoralbio.2021.105060. Epub 2021 Jan 23.
The aim of this study was to investigate the role of microRNA-125a-5p (miR-125a-5p) in macrophages during orthodontic tooth movement (OTM).
Periodontal ligament tissues were collected from patients underwent OTM. Periodontal ligament cells were isolated from periodontal ligament tissues. Periodontal ligament stem cells were isolated from normal human impacted third molars. The miR-125-5p levels were measured by real-time quantitative polymerase chain reaction. The impact of miR-125-5p on macrophage polarization was tested by alizarin red staining assay. The effects of miR-125-5p and E26 transformation-specific variant 6 gene (ETV6) on M1/M2 macrophages phenotype markers were determined by real-time quantitative polymerase chain reaction, western blot, and flow cytometry analyses. The interaction between miR-125-5p and ETV6 was verified using luciferase reporter and RNA immunoprecipitation assays.
Periodontal miR-125a-5p was upregulated under the force. Macrophage polarization facilitated osteogenesis by cocultured system. Moreover, miR-125a-5p was upregulated in macrophages polarized with M2 conditions. MiR-125a-5p downregulation promoted the expression of M1 phenotype markers, while suppressed the expression of M2 markers. Mechanistically, ETV6 was confirmed to be a target of miR-125a-5p. ETV6 overexpression increased the expression of M1 polarized markers, while decreased the expression of M2 polarized markers. Furthermore, ETV6 knockdown reversed the effects of miR-125a-5p inhibitor on both M1 macrophages and M2 macrophages.
Overall, miR-125a-5p facilitates bone healing by targeting ETV6 to promote macrophage M2 polarization.
本研究旨在探讨微小 RNA-125a-5p（miR-125a-5p）在正畸牙齿移动（OTM）过程中对巨噬细胞的作用。
收集接受 OTM 治疗的患者的牙周韧带组织。从牙周韧带组织中分离出牙周韧带细胞。从正常人阻生的第三磨牙中分离出牙周韧带干细胞。通过实时定量聚合酶链反应测量 miR-125-5p 水平。通过茜素红染色试验检测 miR-125-5p 对巨噬细胞极化的影响。通过实时定量聚合酶链反应、western blot 和流式细胞术分析确定 miR-125-5p 和 E26 转化特异性变体 6 基因（ETV6）对 M1/M2 巨噬细胞表型标志物的影响。利用荧光素酶报告和 RNA 免疫沉淀试验验证 miR-125-5p 和 ETV6 之间的相互作用。
在力的作用下，牙周 miR-125a-5p 上调。共培养体系中巨噬细胞极化促进成骨作用。此外，在 M2 条件下极化的巨噬细胞中 miR-125a-5p 上调。miR-125a-5p 下调促进 M1 表型标志物的表达，而抑制 M2 标志物的表达。机制上，ETV6 被证实是 miR-125a-5p 的靶基因。ETV6 过表达增加了 M1 极化标志物的表达，而降低了 M2 极化标志物的表达。此外，ETV6 敲低逆转了 miR-125a-5p 抑制剂对 M1 巨噬细胞和 M2 巨噬细胞的影响。
总的来说，miR-125a-5p 通过靶向 ETV6 促进巨噬细胞 M2 极化来促进骨愈合。
