骨形态发生蛋白-2促进胚胎成骨细胞血管皮内生长因子表达的剂量依赖性效应  

Dose-dependent effect of rhBMP-2 on VEGF expression in fetal mouse osteoblasts

作　　者：傅德皓[1] 杨述华[1] 杨操[1] 李鲲[1] 宁旭[1] 王晶[1] 聂宇[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,湖北武汉430022

出　　处：《创伤外科杂志》2006年第6期541-545,共5页Journal of Traumatic Surgery

基　　金：国家自然科学基金资助项目(30170945;30471753)

摘　　要：目的通过不同剂量重组人骨形态发生蛋白-2(recomb inant hum an bone morphogenetic prote in-2,rhBMP-2)刺激原代培养小鼠胚胎成骨细胞,探讨BMP-2对成骨细胞血管内皮生长因子(vascu lar endothelialgrowth factor,VEGF)表达的影响。方法取19天胎龄小鼠胚胎颅骨成骨细胞进行体外培养,采用RT-PCR、W estern b lot和免疫组化法分别检测不同浓度rhBMP-2对成骨细胞VEGF mRNA及蛋白表达的影响。结果经RT-PCR、W estern b lot及免疫组化法检测表明,VEGF mRNA及蛋白在原代成骨细胞内稳定表达;半定量分析证实,不同rhBMP-2剂量组间VEGF mRNA的表达量呈一定变化规律,随着rhBMP-2浓度的升高,VEGFmRNA的表达量逐渐增加,当rhBMP-2剂量为300ng/m l左右时,VEGF mRNA的表达量最高,超过此剂量,VEGF的表达则有下降趋势。结论VEGF可在原代鼠胚成骨细胞内稳定表达;rhBMP-2可促进VEGF在成骨细胞的表达,并呈一定的剂量依赖性关系,rhBMP-2剂量为300ng/m l左右时,其促进作用最佳。Objective To investigate the effect of different dose of recombinant human bone morphogenetic protein-2 （rhBMP-2） on vascular endothelial growth factor （VEGF） expression in fetal mouse osteoblasts. Methods Calvaria osteoblasts of 19 days old fetal mouse were cultured, the effect of different dose of rhBMP-2 on VEGF mRNA and protein expression in fetal mouse osteoblasts was observed by RT-PCR, western blot and immunocytochemistry. Results The steady-state level of VEGF mRNA and protein expressian in fetal mouse osteoblasts was found by RT-PCR, western blot and immunocytochemistry. The levels of VEGF mRNA increased with an increased stimulation of rhBMP-2, with 300 ng/ml of rhBMP-2. The level of VEGF mRNA and protein expression reached its highest paint,which decreased with more of 300ng/ml of rhBMP-2. Conclusion A steady-state level of VEGF mRNA and protein expression in fetal mouse osteoblasts could be measured by RT-PCR, western blot and immunocytochemistry;rhBMP-2 can promote the expression of VEGF in a dose-dependent manner, the levels of VEGF increased in an apparent biphasic manner with a maximum stimulation at 300ng,/ml of rhBMP-2.

关 键 词：血管内皮生长因子 重组人骨形态发生蛋白-2 成骨细胞 细胞培养 免疫细胞化学 

分 类 号：R392.1[医药卫生—免疫学]