MiR-210-3p 转染对大鼠骨髓基质干细胞成骨活性影响
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1.潍坊医学院;2.北京大学第三医院

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国家自然科学基金项目(面上项目,重点项目,重大项目)


Effect of miR-210-3p regulates the rat bone marrow mesenchymal stem cells on osteogenic activity
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1.Weifang Medical University;2.Peking University Third Hospital

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    摘要:

    [目的] 探讨微小 RNA (microRNA,miR) -210-3p 诱导大鼠骨髓基质干细胞 (Bone Marrow Mesenchymal stem cells, BMMSCs) 治疗骨缺损修复的意义。[方法] 体外培养大鼠 BMMSCs,随机分为空白对照组、miR-210-3p增强 (mimics) 组、miR-210-3p抑制 (inhibitors) 组,分组处理后并诱导其成骨、成脂分化,行茜素红染色检测各组细胞成骨分化情况、油红 O 染色检测各组细胞成脂分化情况;采用 qRT-PCR 检测成骨分化基因碱性磷酸酶 (Alkaline phosphatase,ALP)、骨钙素 (Bone gla protein-2, Bglap-2) mRNA 的表达;以及成脂分化基因过氧化物酶体增殖物激活受体- γ (peroxisome proliferator activated receptor-γ,PPARγ)、脂蛋白脂肪酶基因 (lipoprotein lipase, LPL) mRNA 的表达。通过CCK-8试剂盒、凋亡试剂盒及Transwell实验检测各组细胞增值、凋亡及迁移能力。[结果] miR-210-3p 增强组矿化结节相对比例、 ALP 和 Bglap-2 mRNA 相对表达量增高,脂滴相对比例、 PPARγ 和 LPL mRNA 相对表达量降低;细胞凋亡减少,细胞增值、迁移能力增强; miR-210-3p 抑制组各指标则相反,差异均有统计学意义 (P < 0.05)。[结论] miR-210-3p可促进大鼠 BMMSCs 成骨分化,抑制成脂分化,减少细胞凋亡并促进细胞增值和迁移。 上述结果表明miR-210-3p 结合 BMMSCs 的干细胞疗法对治疗骨缺损具有良好的应用前景。

    Abstract:

    [Objective] This study aimed to investigate the significance of Bone Marrow Mesenchymal stem cells (BMMSCs) in rats induced by micro RNA-210-3p (miR-210-3p) for bone defect regeneration. [Methods] BMMSCs were cultured in vitro and randomly divided into control group, miR-210-3p mimics group and miR-210-3p inhibitors group. Osteogenic differentiation of cells in each group was detected by alizarin red staining, and adipogenic differentiation was detected by oil red O staining. qRT-PCR was used to detect the mRNA expression of Alkaline phosphatase (ALP) and Bone gla protein-2 (Bglap-2) genes of osteogenic differentiation. And a peroxisome proliferator activated receptor γ (PPARγ), lipoprotein lipase (LPL) mRNA expression. Cell proliferation、apoptosis and migration were detected by CCK-8 assay, apoptosis kit, and transwell assay. [Results] In the miR-210-3p mimics group, the relative proportion of mineralized nodules, the relative expression levels of ALP and Bglap-2 mRNA increased. The relative proportion of lipid droplets, PPARγ and LPL mRNA expression were decreased. The cell apoptosis was impeded, and the cell proliferation and migration ability were enhanced. All indicators in miR-210-3p inhibitor group were on the contrary, with statistical significance (P < 0.05). [Conclusions] miR-210-3p can promote osteogenic differentiation, inhibit adipogenic differentiation, reduce cell apoptosis, and enhance proliferation and migration of BMMSCs. These results demonstrated that the combination of miR-210-3p and BMMSCs gene stem cell therapy has a good application prospect in the treatment of bone defect.

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  • 收稿日期:2022-03-15
  • 最后修改日期:2022-05-28
  • 录用日期:2022-06-14
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