miR-195对人骨髓间充质干细胞成骨分化的影响

发布时间：2018-06-01 05:13

  本文选题：骨发育 + 间充质干细胞　； 参考：《中国组织工程研究》2016年45期

【摘要】：背景:既往研究发现,miR-195在人骨髓间充质干细胞成骨分化过程中,表达量显著增加,但是其作用及其机制尚不明确。目的:探索mi R-195对人骨髓间充质干细胞成骨分化的影响及其机制。方法:体外分离、培养人骨髓间充质干细胞,通过碱性磷酸酶活性测定试剂盒、蛋白免疫和实时定量反转录PCR(q RT-PCR)检测在人骨髓间充质干细胞诱导分化过程中,碱性磷酸酶活性和骨分化特异基因Runx2和osteopontin表达水平的变化,以及miR-195和它的潜在靶SMAD7表达量的变化;通过脂质体转染构建miR-195低表达的人骨髓间充质干细胞,研究miR-195对人骨髓间充质干细胞成骨分化的影响。利用荧光素酶报告基因实验检测miR-195对SMAD7的靶向作用。结果与结论:(1)分离培养的人骨髓间充质干细胞具有优良的体外成骨分化能力;(2)miR-195表达量随人骨髓间充质干细胞诱导分化时间的增加而升高,SMAD7则相反。(3)mi R-195能够促进人骨髓间充质干细胞成骨分化;(4)荧光素酶实验证实SMAD7是miR-195的直接靶,SMAD7过表达抑制人骨髓间充质干细胞成骨分化;(5)结果提示,miR-195通过靶向SMAD7促进人骨髓间充质干细胞成骨分化。
[Abstract]:Background: previous studies have found that miR-195 increased significantly during osteogenic differentiation of human bone marrow mesenchymal stem cells, but its role and mechanism are unclear. Aim: to investigate the effect of mi R 195 on osteogenic differentiation of human bone marrow mesenchymal stem cells (BMSCs) and its mechanism. Methods: human bone marrow mesenchymal stem cells were isolated and cultured in vitro. The differentiation of human bone marrow mesenchymal stem cells was detected by alkaline phosphatase assay kit, protein immunoassay and real-time reverse transcription PCR(q RT-PCR. The changes of alkaline phosphatase activity and the expression level of bone differentiation specific gene Runx2 and osteopontin, as well as the changes of miR-195 and its potential target SMAD7 expression, and the construction of human bone marrow mesenchymal stem cells with low expression of miR-195 by liposome transfection. To study the effect of miR-195 on osteogenic differentiation of human bone marrow mesenchymal stem cells. Luciferase reporter gene experiment was used to detect the targeting effect of miR-195 on SMAD7. Results and conclusion Human bone marrow mesenchymal stem cells (BMSCs) isolated and cultured have excellent osteogenic differentiation ability in vitro. The expression of miR-195 increases with the increase of differentiation time of human bone marrow mesenchymal stem cells. On the contrary, SMAD7 can promote human osteogenic differentiation. Luciferase assay confirmed that SMAD7 was a direct target of miR-195 for inhibiting osteogenic differentiation of human bone marrow mesenchymal stem cells (BMSCs). The results suggested that SMAD7-195 could promote osteogenic differentiation of human bone marrow mesenchymal stem cells by targeting SMAD7.
【作者单位】： 河南省中医院;
【分类号】：R329.2
，

本文编号：1962966