miR-129-1-3p通过BMP2/SMAD1信号通路抑制人骨髓间充质干细胞成骨分化的研究

doi:10.13591/j.cnki.kqyx.2025.06.004

摘要/Abstract

摘要：

目的探讨miR-129-1-3p对人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,hBMSCs)成骨分化的作用及相关机制研究。方法构建空白对照(NC组)、miR-129-1-3p模拟物(Mimic组)、miR-129-1-3p抑制物(Inhibitor组)及其相应的阴性对照组(Mimic-NC组、Inhibitor-NC组)并转染至hBMSCs。碱性磷酸酶和茜素红染色观察钙盐矿化结节形成能力,qRT-PCR检测miR-129-1-3p及成骨分化标志物表达水平,western blot检测骨形态发生蛋白2(bone morphogenetic protein 2,BMP2)、SMAD1及磷酸化SMAD1蛋白表达水平。结果转染后,相较于Mimic-NC组,Mimic组miR-129-1-3p表达水平显著升高(P<0.05),矿化结节显著减少,BMP2、Runt相关转录因子2(Runt-related transcription factor 2,RUNX2)、骨钙素(osteocalcin,OCN)mRNA表达水平显著下调(P<0.05),BMP2和磷酸化SMAD1蛋白表达水平显著下调(P<0.05)。Inhibitor组相较于Inhibitor-NC组矿化结节明显增多,BMP2、RUNX2、OCN mRNA表达显著上调(P<0.05),BMP2和磷酸化SMAD1蛋白表达水平显著上调(P<0.05)。结论 miR-129-1-3p可能通过调控BMP2/SMAD1信号通路抑制人骨髓间充质干细胞成骨分化。

关键词: miR-129-1-3p, BMP2/SMAD1信号通路, 人骨髓间充质干细胞, 成骨分化, 骨质疏松

Abstract:

Objective To investigate the effect of miR-129-1-3p on the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) and its potential mechanism. Methods Negative control group, the miR-129-1-3p mimic group, the miR-129-1-3p inhibitor group and the corresponding negative control were constructed and transfected into hBMSCs. The formation of calcium-mineralized nodules was observed by alkaline phosphatase staining and alizarin red S staining. The expression levels of miR-129-1-3p and osteogenic differentiation markers were detected by qRT-PCR. Western blot detected the protein expressions of bone morphogenetic protein 2 (BMP2), SMAD1 and p-SMAD1. Results After transfection, the expression level of miR-129-1-3p in mimic group was significantly increased (P<0.05), the number of mineralized nodules was significantly decreased, and the expression levels of BMP2, Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) mRNA were significantly down-regulated (P<0.05). BMP2 and p-SMAD1 protein were also significantly down-regulated (P<0.05) compared with Mimic-NC group. The expression levels of BMP2, RUNX2, OCN mRNA were significantly up-regulated in inhibitor group (P<0.05) compared with Inhibitor-NC group. BMP2 and p-SMAD1 protein were significantly up-regulated in inhibitor group (P<0.05). Conclusion miR-129-1-3p can inhibit the osteogenic differentiation of human bone marrow mesenchymal stem cells by suppressing BMP2/SMAD1 signaling pathway.

Key words: miR-129-1-3p, BMP2/SMAD1 signaling pathway, human bone marrow mesenchymal stem cells, osteogenic differentiation, osteoporosis

中图分类号:

R780.2

耿铭珠, 穆文清, 邱琳, 张玮. miR-129-1-3p通过BMP2/SMAD1信号通路抑制人骨髓间充质干细胞成骨分化的研究[J]. 口腔医学, 2025, 45(6): 418-423.

GENG Mingzhu, MU Wenqing, QIU Lin, ZHANG Wei. miR-129-1-3p inhibits osteogenic differentiation of human bone marrow mesenchymal stem cells via BMP2/SMAD1 signaling pathway[J]. Stomatology, 2025, 45(6): 418-423.

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基因名称	引物序列(5'→3')
RUNX2-F	GACGAGGCAAGAGTTTCACC
RUNX2-R	GGTTCCCGAGGTCCATCTAC
BMP2-F	TTCGGCCTGAAACAGAGACC
BMP2-R	CCTGAGTGCCTGCGATACAG
OCN-F	CTACCTGTATCAA
OCN-R	GGATTGAGCTCACACACCT
miR-129-1-3p RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCA-CTGGATACGACATACTT
miR-129-1-3p-F	ACCATTCAAAGCCCTTACCCCAA
miR-129-1-3p-R	ATCCAGTGCAGGGTCCGAGG
GAPDH-F	CAATGACCCCTTCATTGACC
GAPDH-R	TTGATTTTGGAGGGATCTCG