Brg1調(diào)控重組人骨形態(tài)發(fā)生蛋白2誘導成骨細胞分化
發(fā)布時間:2018-05-22 09:03
本文選題:成骨 + 染色質(zhì)重塑。 參考:《暨南大學》2011年碩士論文
【摘要】:目的:基于原代培養(yǎng)的小鼠顱骨細胞,探索Brg1基因在骨形態(tài)發(fā)生蛋白2誘導成骨細胞分化過程中的調(diào)控機制。 方法:采用膠原酶消化法進行小鼠顱骨成骨細胞的原代培養(yǎng);分別用0,50,200μg/L的重組人骨形態(tài)發(fā)生蛋白2誘導原代培養(yǎng)的成骨細胞的分化,摸索骨形態(tài)發(fā)生蛋白2的最佳作用劑量;實時熒光定量PCR和Western blot進行骨形態(tài)發(fā)生蛋白2對Brg1的作用時間的動力學分析;實時熒光定量PCR和鈣鈷染色法檢測敲除Brgl對骨形態(tài)發(fā)生蛋白2誘導的成骨分化的影響;構(gòu)建Dlx5腺病毒重組表達載體,實時熒光定量PCR和鈣鈷染色法檢測Brg1在骨形態(tài)發(fā)生蛋白2誘導的成骨分化過程中對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可誘導原代培養(yǎng)小鼠成骨細胞分化,200μg/L劑量有著較好的誘導分化效果;重組人骨形態(tài)發(fā)生蛋白2可誘導Brg1基因轉(zhuǎn)錄水平和翻譯水平表達水平上調(diào);敲除Brgl可抑制重組人骨形態(tài)發(fā)生蛋白2誘導的成骨分化;Brg1能夠調(diào)控Dlx5的表達水平。說明Brg1通過調(diào)控Dlx5的表達水平調(diào)控重組人骨形態(tài)發(fā)生蛋白2誘導的小鼠成骨細胞的分化。
[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.
【學位授予單位】:暨南大學
【學位級別】:碩士
【學位授予年份】:2011
【分類號】:R329
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