本文選題：骨發(fā)育 + 間充質(zhì)干細(xì)胞　； 參考：《中國(guó)組織工程研究》2016年45期

【摘要】：背景:既往研究發(fā)現(xiàn),miR-195在人骨髓間充質(zhì)干細(xì)胞成骨分化過(guò)程中,表達(dá)量顯著增加,但是其作用及其機(jī)制尚不明確。目的:探索mi R-195對(duì)人骨髓間充質(zhì)干細(xì)胞成骨分化的影響及其機(jī)制。方法:體外分離、培養(yǎng)人骨髓間充質(zhì)干細(xì)胞,通過(guò)堿性磷酸酶活性測(cè)定試劑盒、蛋白免疫和實(shí)時(shí)定量反轉(zhuǎn)錄PCR(q RT-PCR)檢測(cè)在人骨髓間充質(zhì)干細(xì)胞誘導(dǎo)分化過(guò)程中,堿性磷酸酶活性和骨分化特異基因Runx2和osteopontin表達(dá)水平的變化,以及miR-195和它的潛在靶SMAD7表達(dá)量的變化;通過(guò)脂質(zhì)體轉(zhuǎn)染構(gòu)建miR-195低表達(dá)的人骨髓間充質(zhì)干細(xì)胞,研究miR-195對(duì)人骨髓間充質(zhì)干細(xì)胞成骨分化的影響。利用熒光素酶報(bào)告基因?qū)嶒?yàn)檢測(cè)miR-195對(duì)SMAD7的靶向作用。結(jié)果與結(jié)論:(1)分離培養(yǎng)的人骨髓間充質(zhì)干細(xì)胞具有優(yōu)良的體外成骨分化能力;(2)miR-195表達(dá)量隨人骨髓間充質(zhì)干細(xì)胞誘導(dǎo)分化時(shí)間的增加而升高,SMAD7則相反。(3)mi R-195能夠促進(jìn)人骨髓間充質(zhì)干細(xì)胞成骨分化;(4)熒光素酶實(shí)驗(yàn)證實(shí)SMAD7是miR-195的直接靶,SMAD7過(guò)表達(dá)抑制人骨髓間充質(zhì)干細(xì)胞成骨分化;(5)結(jié)果提示,miR-195通過(guò)靶向SMAD7促進(jìn)人骨髓間充質(zhì)干細(xì)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.
【作者單位】： 河南省中醫(yī)院;
【分類號(hào)】：R329.2
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本文編號(hào)：1962966