發(fā)布時(shí)間：2018-09-07 19:24

【摘要】：目的:研究Runx1在骨形態(tài)發(fā)生蛋白9(bone morphogenetic proteins 9,BMP9)誘導(dǎo)小鼠胚胎成纖維細(xì)胞(mouse embryonic fibroblasts,MEFs)成骨分化中的作用。方法:用BMP9腺病毒感染MEFs細(xì)胞,利用RT-PCR和Western blot分別在mRNA和蛋白質(zhì)水平檢測(cè)Runx1的內(nèi)源性表達(dá);構(gòu)建過(guò)表達(dá)Runx1的重組腺病毒Ad-Runx1,并在mRNA和蛋白質(zhì)水平驗(yàn)證Ad-Runx1的效果;用Ad-Runx1和BMP9條件培養(yǎng)基共處理MEFs,檢測(cè)成骨早期指標(biāo)堿性磷酸酶(ALP)染色和活性,茜素紅S染色檢測(cè)成骨晚期指標(biāo)鈣鹽沉積;RT-PCR和Western blot分別檢測(cè)成骨關(guān)鍵轉(zhuǎn)錄因子Runx2在mRNA和蛋白質(zhì)水平的變化。結(jié)果:Ad-BMP9處理MEFs細(xì)胞后,可使Runx1在mRNA和蛋白質(zhì)水平表達(dá)上調(diào);構(gòu)建的Ad-Runx1處理MEFs后,可使Runx1在mRNA和蛋白質(zhì)水平表達(dá)上調(diào);Ad-Runx1處理BMP9誘導(dǎo)的MEFs細(xì)胞后,增強(qiáng)了ALP活性和鈣鹽沉積以及Runx2在mRNA和蛋白質(zhì)水平的表達(dá)。結(jié)論:Runx1可以促進(jìn)BMP9誘導(dǎo)的間充質(zhì)干細(xì)胞MEFs的成骨分化。
[Abstract]:Aim: to investigate the role of Runx1 in osteogenic differentiation of mouse embryonic fibroblasts (mouse embryonic fibroblasts,MEFs) induced by bone morphogenetic protein 9 (bone morphogenetic proteins 9 (BMP9). Methods: BMP9 adenovirus was used to infect MEFs cells, and RT-PCR and Western blot were used to detect the endogenous expression of Runx1 at the mRNA and protein levels, respectively, and the recombinant adenovirus Ad-Runx1, expressing Runx1 was constructed, and the effect of Ad-Runx1 was verified at mRNA and protein levels. (ALP) staining and activity of alkaline phosphatase (ALP) were detected by co-treatment of MEFs, with Ad-Runx1 and BMP9 conditioned medium in the early stage of osteogenesis. Alizarin red S staining was used to detect the changes of Runx2 in mRNA and protein levels in late osteogenic stage by RT-PCR and Western blot, respectively. Results the expression of Runx1 and protein in MEFs cells were up-regulated after treated with: Ad-BMP9, and Runx1 was up-regulated at mRNA and protein levels in BMP9 induced MEFs cells after MEFs was treated with constructed Ad-Runx1. Enhanced ALP activity, calcium deposition and Runx2 expression at mRNA and protein levels. Conclusion 1: Runx 1 can promote the osteogenic differentiation of mesenchymal stem cells (MEFs) induced by BMP9.
【作者單位】： 重慶醫(yī)科大學(xué)檢驗(yàn)醫(yī)學(xué)院重慶醫(yī)科大學(xué)檢驗(yàn)醫(yī)學(xué)教育部重點(diǎn)實(shí)驗(yàn)室重慶市重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(31071304,81272006)
【分類(lèi)號(hào)】：R329.2

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