骨形态发生蛋白-2促进鼠胚成骨细胞血管内皮生长因子表达的机制研究  

VEGF expression induced by rbBMP-2 in fetal mouse osteoblasts

作　　者：傅德皓[1] 杨述华[1] 刘国辉[1] 李进[1] 许伟华[1] 梅荣成[1] 郭晓东[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院骨科,武汉430022

出　　处：《中华创伤骨科杂志》2007年第10期964-966,共3页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金资助项目（30471753,30470483）

摘　　要：目的探讨重组人骨形态发生蛋白-2（rhBMP-2）促进鼠胚成骨细胞血管内皮生长因子（VEGF）表达的机制。方法取19d胎龄的小鼠胚胎颅骨成骨细胞进行体外培养，在加入rhBMP-2诱导成骨的同时，分别加入放线菌素D、放线菌酮和羟（基）脲三种药物，RT—PCR法检测VEGFmRNA表达的变化。结果应用放线菌素D阻断基因转录后，可降低成骨细胞VEGFmRNA的表达，并可完全抑制BMP-2对VEGF表达的促进作用。用放线菌酮阻断成骨细胞的蛋白质合成后，可增加成骨细胞VEGFmRNA的表达，但不能完全抑制BMP-2对VEGFmRNA表达的促进作用。应用羟（基）脲阻断DNA合成后，可抑制BMP-2对VEGF分泌的促进作用。结论rhBMP-2主要通过促进VEGFmRNA的转录来促进鼠胚成骨细胞VEGF基因表达，并与BMP-2的促有丝分裂活性密切相关，新蛋白的合成并不是促进VEGFmRNA表达的必要条件。Objective To investigate the mechanism of stimulation of vascular endothelial growth factor （VEGF） expression in fetal mouse osteoblasts by the recombinant human bone morphogenetic protein-2 （rhBMP-2）. Methods Calvaria osteoblasts of fetal mice of 19 days were cultured. Effects of actinomycin D, cycloheximide, and hydroxyurea on VEGF mRNA expression level in fetal mouse osteoblasts with presence or absence of rhBMP-2 were observed with reverse transcription-polymerase chain reaction （RT-PCR） . Results Inhibition of transcription by actinomycin D reduced VEGF transcription in control cells and almost completely prevented the stimulation of VEGF expression by rhBMP-2. Cycloheximide increased VEGF expression in control cells but it did not abolish the rhBMP-2 effect. Inhibition of DNA synthesis blocked the stimulatory effects of rhBMP-2 on VEGF gene expression. Conclusions The observation suggests that the observed enhancement of VEGF expression induced by rhBMP-2 is dependent on new transcription and cell replication （the mitogenic activity of rhBMP-2）. The results indicate that de novo protein synthesis might not be essential for the increase of VEGF mRNA.

关 键 词：生长调节素类 骨形态发生蛋白质类 成骨细胞 细胞培养 鼠科 

分 类 号：R686[医药卫生—骨科学]