| lncRNA H19促进脐带间充质干细胞成骨分化的研究 |
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投稿时间：2023-07-08 修订日期：2024-01-15
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| 期刊信息：《中国骨伤》年，第卷，第期，第-页 |
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| 中文摘要:目的：观察长链非编码RNA H19（long non-coding RNAs lncRNA H19，lncRNA H19，以下简称H19）转染人脐带间充质干细胞后对多个成骨成血管因子表达的影响及其成骨性能，为今后合成血供良好的骨缺损填充物提供前期研究基础。方法：生物信息学确定H19可以调控与骨形成相关的微小RNA（MicroRNA，miRNA）及这些miRNAs可调控的成骨成血管因子，最后将目标锁定为miR-137-3p、miR-383-5p和它们可以调控的骨涎蛋白（Human bone saliva protein，IBSP）、Runt结构域基因家族的Runx2（ runt-related transcription factor 2）、和成纤维细胞生长因子5（Fibroblast growth factors 5，FGF5）。体外培养人脐带间充质干细胞。然后将lncRNA H19转染致人脐带间充质干细胞，反转录-聚合酶链式扩增技术（Reverse Transcription-Polymerase Chain Reaction，RT-PCR）法观察转染后细胞中miR-137-3p、miR-383-5p表达丰度及western blot法观察IBSP、RUNX2、和FGF5表达情况，碱性磷酸酶和茜素红染色观察转染H19后细胞的成骨性能。验证lncRNA H19转染人脐带间充质干细胞是否可以通过降低miR-137-3p、miR-383-5p的表达，从而提高成骨成血管因子IBSP、RUNX2、和FGF5的表达，并确认转染lncRNA H19可以促进人脐带间充质干细胞的成骨作用。结果：转染lncRNA H19的人脐带间充质干细胞内miR-137-3p、miR-383-5p表达下调，而RUNX2 、IBSP和FGF5的表达均明显升高，分别由转染前0.533±0.058，0.663±0.058，0.500±0.100升高至转然后1.300±1.100，1.233±0.058，0.967±0.115，差异具有显著统计学意义（P=0.000，P=0.000，P=0.006）。且脐带间充质干细胞转染H19后成骨性能更好。结论：转染lncRNA H19的人脐带间充质干细胞可以促进某些成骨成血管因子的表达且成骨性能提高，为今后研制一种包含有lncRNA H19的组织工程骨，促进该组织工程骨的成骨及成血管性能提供可能。 |
| 【关键词】lncRNA H19 人脐带间充质干细胞 成骨性能 RUNX2 IBSP FGF5 |
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| Experimental study on lncRNA H19 promoting the expression of osteogenic and angiogenesis factors in bone marrow mesenchymal stem cells |
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| ABSTRACT ABSTRACT Objective To observe the effect of lncRNA H19 transfecting human umbilical cord mesenchymal stem cells on the expression of multiple osteogenic angiogenesis factors and osteogenic property,which will provide a preliminary research basis for the future synthesis of bone defect fillers that can simultaneously promote the high expression of multiple osteogenic angiogenesis factors.Methods biological information website were used to identify lncRNA H19-regulated bone formation-related miRNAs and related osteogenic-angiogenic factors. MiR-137-3p, miR-383-5p and factors IBSP, Runx2, and FGF5 were selected. Bone marrow mesenchymal stem cells were cultured in vitro. Then lncRNA H19 was transfected into bone marrow mesenchymal stem cells, and the expression abundance of miR-137-3p, miR-383-5p in the transfected cells was observed by PCR method, and expression of IBSP, Runx2, and FGF5 was observed by western blot. It was confirmed that lncRNA H19 transfection of bone marrow mesenchymal stem cells can reduce the expression of miR-137-3p, miR-383-5p, thereby increasing expression level of the osteogenic angiogenic factors IBSP, Runx2 and FGF5.It was confirmed that transfection of lncRNA H19 promoted the osteogenesis and angiogenesis of bone marrow mesenchymal stem cells.Results The expression of miR-137-3p and miR-383-5p in BMSCs transfected with lncRNA H19 was down-regulated, while the expressions of RUNX2 , IBSP and FGF5 were significantly increased from 0.533±0.058，0.663±0.058，0.500±0.100 to 1.300±1.100，1.233±0.058，0.967±0.115 after transfection. The difference was statistically significant（P=0.000，P=0.000，P=0.006）.Conclusions HUCB-MSCS transfected with lncRNA H19 can promote the expression of some osteogenic angiogenic factor.Bone marrow mesenchymal stem cells transfected with lncRNA H19 are promising and may have better osteogenic and angiogenic properties in the future. |
| KEY WORDS lncRNA H19 Bone marrow mesenchymal stem cells Osteogenic and angiogenic properties RUNX2 IBSP |
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