| 摘要: |
| 目的：研究miR- 122-5p对人骨髓间充质干细胞hBMSCs细胞成骨分化诱导的作用分子机制。方法：运用茜素红 S、 碱性磷酸酶活性ALP染色试剂盒检测hBMSCs成骨分化的钙结节与成骨活性；使用双荧光素酶报告基因实验验证miR - 122- 5p与 BRCC3之间的靶向关系；运用 qRT-PCR分别检测Runx2、Osterix、wnt3a、β -catenin、GSK3- β 、miR- 122-5 以及BRCC3基因 的表达情况；Western 印迹检测 β-catenin蛋白的表达量。结果：过表达miR- 122-5p对人骨髓间充质干细胞hBMSCs细胞成骨分 化诱导21天后的钙结节与干细胞成骨活性均有明显促进作用，对成骨分化中wnt/β-catenin信号通路主要转录因子Runx2、Os- terix及wnt3a、β-catenin、GSK3- β 主要基因均激活作用，过表达BRCC3对干细胞成骨分化中wnt/β-catenin信号通路主要转录因 子 Runx2、Osterix 及 wnt3a、β -catenin、GSK3- β 主要基因有抑制作用；并且通过双荧光素酶报告基因与回复性实验确定miR- 122-5p与 BRCC3之间的靶向关系，差异均有统计学意义 (P<0. 05) 。结论：miR- 122-5p靶向抑制BRCC3 的表达诱导hBMSCs 细胞通过wnt/β-catenin通路进行成骨分化诱导。 |
| 关键词: miR- 122-5p 人骨髓间充质干细胞 成骨分化 wnt/β-catenin信号通路 |
| DOI：10. 19787/j.issn.2097-3128.2024.01.006 |
| 投稿时间：2023-02-23修订日期：2023-10-13 |
| 基金项目:佛山市科技局医学类科技攻关项目(NO. 1920001001298) |
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| Study on The Molecular Mechanism of MiR- 122-5p Targeted Regulation of Osteogenic Differentiation Induced by Human Bone Marrow Mesenchymal Stem Cells |
| CAI Li-xiong, MIAO Zhong-lu |
| (Foshan hospital of TCM, Foshan, Guangdong 528051) |
| Abstract: |
| Objective: To explore the role and mechanism of miR- 122-5p on the osteogenic differentiation of hBMSCs. Methods: we investigated the role of miR- 122-5p in osteogenic differentiation by alkaline phosphatase staining and Alizarin red staining，Dual luciferase reporter gene assay was used to verify the targeting relationship between miR- 122-5p and BRCC3, Expression of Runx2、Osterix、wnt3a、β -catenin、GSK3- β、miR- 122-5 and BRCC3 mRNA in hBMSCs were detected by quantitative reverse transcription polymerase chain reaction(RT- qPCR). Western blot was used to de- tect the expression of β -catenin protein. Results: The over expression of MiR- 122-5p significantly promoted the activation of alkaline phosphatase (ALP) and formation of calcified nodules in osteoblasts, The mRNA expressions of β-catenin、Runx2、Osterix、wnt3a、GSK3-β and the protein expres- sion of β -catenin were significantly higher than those in control group. The dual-luciferase reporter gene experiment confirmed that BRCC3 was the downstream target gene of miR- 122-5p. At the same time, inhibited osteogenic differentiation was also observed after BACC3 overexpression, the ex- pression levels ofβ-catenin、Runx2、Osterix、wnt3a、GSK3- β were inhibited. Conclusion: miR- 122-5p can promote osteogenic differentiation of hBM- SCs, and targeting BACC3affects the activation of the wnt/β-catenin signaling pathway and regulates the process of osteoporosis treatment. |
| Key words: MiR- 122-5p BRCC3 wnt/β-catenin signaling osteoporosis |
