本文選題：成骨 + 染色質(zhì)重塑�。� 參考：《暨南大學(xué)》2011年碩士論文

【摘要】：目的：基于原代培養(yǎng)的小鼠顱骨細(xì)胞,探索Brg1基因在骨形態(tài)發(fā)生蛋白2誘導(dǎo)成骨細(xì)胞分化過程中的調(diào)控機(jī)制。 方法：采用膠原酶消化法進(jìn)行小鼠顱骨成骨細(xì)胞的原代培養(yǎng)；分別用0,50,200μg/L的重組人骨形態(tài)發(fā)生蛋白2誘導(dǎo)原代培養(yǎng)的成骨細(xì)胞的分化,摸索骨形態(tài)發(fā)生蛋白2的最佳作用劑量；實(shí)時(shí)熒光定量PCR和Western blot進(jìn)行骨形態(tài)發(fā)生蛋白2對(duì)Brg1的作用時(shí)間的動(dòng)力學(xué)分析；實(shí)時(shí)熒光定量PCR和鈣鈷染色法檢測敲除Brgl對(duì)骨形態(tài)發(fā)生蛋白2誘導(dǎo)的成骨分化的影響；構(gòu)建Dlx5腺病毒重組表達(dá)載體,實(shí)時(shí)熒光定量PCR和鈣鈷染色法檢測Brg1在骨形態(tài)發(fā)生蛋白2誘導(dǎo)的成骨分化過程中對(duì)Dlx5的調(diào)控作用。 結(jié)果與結(jié)論：用自行合成的重組人骨形態(tài)發(fā)生蛋白2可誘導(dǎo)原代培養(yǎng)小鼠成骨細(xì)胞分化,200μg/L劑量有著較好的誘導(dǎo)分化效果；重組人骨形態(tài)發(fā)生蛋白2可誘導(dǎo)Brg1基因轉(zhuǎn)錄水平和翻譯水平表達(dá)水平上調(diào)；敲除Brgl可抑制重組人骨形態(tài)發(fā)生蛋白2誘導(dǎo)的成骨分化；Brg1能夠調(diào)控Dlx5的表達(dá)水平。說明Brg1通過調(diào)控Dlx5的表達(dá)水平調(diào)控重組人骨形態(tài)發(fā)生蛋白2誘導(dǎo)的小鼠成骨細(xì)胞的分化。
[Abstract]:Objective: To explore the regulatory mechanism of Brg1 gene in osteoblast differentiation induced by bone morphogenetic protein 2 based on primary cultured mouse cranial cells.
Methods: the primary culture of mouse skull osteoblasts was cultured with collagenase digestion; the differentiation of primary cultured osteoblasts was induced by recombinant human bone morphogenetic protein 2 0,50200 micron g/L, and the optimal dosage of bone morphogenetic protein 2 was explored. Real time fluorescence quantitative PCR and Western blot were used for bone morphogenetic protein 2 pairs. Dynamic analysis of the action time of Brg1; real-time fluorescence quantitative PCR and calcium cobalt staining to detect the effect of knockout Brgl on osteogenic differentiation induced by bone morphogenetic protein 2; construct Dlx5 adenovirus recombinant expression vector, real-time fluorescent quantitative PCR and calcium cobalt staining method to detect Brg1 in the osteogenic differentiation induced by bone morphogenetic protein 2 to D The regulatory role of LX5.
Results and conclusion: the self synthesized recombinant human bone morphogenetic protein 2 could induce the differentiation of osteoblasts in the primary culture of mice. The dose of 200 g/L had a better induction of differentiation. Recombinant human bone morphogenetic protein 2 could induce the up-regulation of Brg1 gene transcription level and the level of translation level, and knockout Brgl could inhibit the morphology of recombinant human bone. The osteogenic differentiation induced by protein 2; Brg1 can regulate the expression level of Dlx5. It shows that Brg1 regulates the differentiation of mouse osteoblasts induced by recombinant human bone morphogenetic protein 2 by regulating the expression level of Dlx5.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R329

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