骨形态发生蛋白-2对原代鼠胚成骨细胞血管内皮生长因子表达的影响  

Effect of human bone morphogenetic protein-2 on vascular endothelial growth factor expression in fetal mouse osteoblasts

作　　者：傅德皓[1] 杨述华[1] 肖宝钧[1] 杨操[1] 叶哲伟[1] 李鲲[1] 宁旭[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》2006年第8期751-754,共4页Chinese Journal of Orthopaedic Trauma

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(rhBMP-2)刺激原代培养的小鼠胚胎成骨细胞,探讨BMP-2对成骨细胞血管内皮生长因子(VEGF)表达的影响及规律。方法取19 d胎龄的小鼠胚胎颅骨成骨细胞进行体外培养,采用RT-PCR法或Western blot法染色法检测不同浓度rhBMP-2、不同作用时间下对成骨细胞VEGF mRNA表达的影响及规律。结果RT-PCR法检测到VEGF mRNA在原代成骨细胞内稳定表达；半定量分析表明,随着rhBMP-2浓度的升高,VEGF mRNA的表达量逐渐增加,当rhBMP-2浓度为300 ng/mL左右时,VEGF mRNA的表达量最高,超过此浓度,VEGF的表达则有下降趋势；应用终浓度rhBMP-2(300 ng/mL)刺激成骨细胞时,不同作用时间下VEGF mRNA的表达量呈双时相,分别在3 h和24 h时出现高峰,48 h时VEGF mRNA的表达仍保持较高水平。除去各自对照组的影响,各时间点VEGF表达的相对值[(rhBMP-2组/对照组)比值]亦呈时间依赖性关系,6 h时明显升高为对照组的2．5倍(P＜0．05),36 h时约升高为对照组的2倍。结论VEGF可在原代鼠胚成骨细胞内稳定表达；rhBMP-2在体外可刺激原代骨细胞VEGF的表达,并呈一定的浓度和时间依赖性关系。Objective To investigate effects of different doses of recombinant human bone morphogenetic protein-2 （rhBMP-2） on vascular endothelial growth factor （VEGF） expression in fetal mouse osteoblasts. Methods Calvaria osteoblasts of fetal mice of 19 days were cultured. The effects of rhBMP-2 at different doses and different action times on VEGF expression patterns in fetal mouse osteoblasts were observed with reverse transcription-polymerase chain reaction （RT-PCR） and Western blot staining. Results In the present study, RT-PCR detected a steady expression of VEGF mRNA in the control fetal mouse osteoblasts. The levels of VEGF mRNA increased in an apparent biphasic manner with a maximum stimulation （about 2-fold above the control, P 〈 0. 05） in both VEGF mRNA species observed at 300 ng/mL of rhBMP-2. After 48 h of rhBMP-2 treatment, the VEGF mRNA levels approached those in the control. The VEGF mRNA levels appeared to be biphasic in rhBMP-2-treated cultures, showing peak induction at 3 and 24 h and remaining elevated at 48 h. Compared with the individual control value at each time point, an apparent maximum increase （about 2.5-fold above the control, P 〈 0. 05） occurred at 6 h. The second peak induction, about 2-fold above that in the control, occurred at about 36 h. Conclusions The expression of VEGF mRNA is steady in the control fetal mouse osteoblasts. RhBMP-2 can promote the expression of VEGF in dose-dependent and time-dependent manners.

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 原代培养 

分 类 号：R68[医药卫生—骨科学]