中文摘要：

背景：Brgl是依赖ATP的染色质改变复合物的核心催化弧基，该亚基在基因的转录调控、复制、重组，骨骼肌的分化、抑制肿瘤的发生等活动中起着重要的作用。目的：探索Brgl基因在骨形态发生蛋白2诱导成骨细胞分化过程中的调控机制。方法：采用胶原酶消化法进行小鼠颅骨成骨细胞的原代培养；分别用0，50，200pg／L的重组人骨形态发生蛋白2诱导原代培养的成骨细胞的分化，摸索骨形态发生蛋白2的最佳作用剂量：实时荧光定量PCR和Westernblot进行骨形态发生蛋白2对Brgl的作用时间的动力学分析：实时荧光定量PCR和钙钴染色法检测敲除Brgl对骨形态发生蛋白2诱导的成骨分化的影响；构建DIx5腺病毒重组表达载体，实时荧光定量PCR和钙钴染色法检测Brgl在骨形态发生蛋白2诱导的成骨分化过程中对Dlx5的调控作用。结果与结论：用自行合成的重组人骨形态发生蛋白2可诱导原代培养小鼠成骨细胞分化，200pg／L剂量有着较好的诱导分化效果；重组人骨形态发生蛋白2可诱导Brgl基因转录水平和翻译水平表达水平上调；敲除Brgl可抑制熏组人骨形态发生蛋白2诱导的成骨分化：Brgl能够调控DIx5的表达水平。说明Brgl通过调控DIx5的表达水平调控重组入骨形态发生蛋白2诱导的小鼠成骨细胞的分化。

英文摘要：

BACKGROUND： Brgl is an ATPase subunit of the SWI/SNF complex and plays an important role in the regulation of gene transcription, replication and recombination, skeletal muscle differentiation and inhibition carcinogenesis. OBJECTIVE： To explore the Brgl regulation mechanism in bone morphogenetic protein 2 （BMP-2）-induced osteogenic differentiation. METHODS： Mouse calvarial osteoblasts were primarily cultured through collagenase digestion and induced differentiation using 0, 50 or 200 pg/L BMP2 to select the optimal dose. Transcription time dynamics analysis of Brgl was enforced by RT-PCR and Western blot after BMP-2 treatments; ALP staining was used to evaluate the capability of osteogenic differentiation after BMP-2 treatments. The role of BMP-2 on Brgl mRNA expression level was detected by quantitative RT-PCR and the protein level was detected by Western Blot. Effect of Brgl on BMP-2-induced differentiation was analyzed by Brgl-siRNA; Ad-DIx5 was obtained by recombinant adenovirus. RESULTS AND CONCLUSION： BMP-2 could induce the osteogenic differentiation of primarily cultured mouse osteoblast, and the optimal BMP2 induction condition was 200 pg/L. The expression of Brgl was up-regulated by BMP-2 in primarily cultured mouse osteoblasts. BMP-2 induced the osteogenic differentiation was inhibited by Brgl-siRNA. The expression of DIx5 was regulated by Brgl. Brgl could regulate the BMP-2-induced osteogenic differentiation of primarily cultured mouse osteoblast through regulating the expression of DIx5.