骨折愈合过程中血管内皮生长因子基因扩增及蛋白表达研究

The study of VEGF gene amplification and protein expression in the process of fracture healing

目的 探讨骨折愈合过程中血管内皮细胞生长因子 (VEGF)基因转录及蛋白表达特点。方法 利用 3 5只大耳白兔制作桡骨骨折模型 ,于不同时相点将兔处死。从骨折端提取总RNA及蛋白样品 ,并进行VEGF基因逆转 聚合酶链反应 (RT PCR)扩增和蛋白免疫印记检测。结果 从伤后各时相点骨折端提取的总RNA中扩增到 2 3 0bp的目的基因片段 ;利用免疫印记法进行的VEGF多克隆抗体杂交检测到相对分子质量为 45× 10 3 的蛋白条带。结论 骨折后 72h～ 3周为VEGF表达的高峰期 ,其表达亚型为VEGF165。

Objective To study the VEGF gene transcription and characteristic of VEGF protein expression.Methods Thirty five rabbits were used to made radius fracture models and sacrificed at intervals of different time.The total RNA and protein were isolated from the fractured ends,and were used for RT PCR amplification of VEGF gene and protein detection by Western blot.Results The aimed gene fragment of 230?bp was amplified from the total RNA isolated from the fractured ends of every time interval and the protein molecular weight was about 45×10 3 detected in the isolated protein from fractured ends with polyclonal antibody by Western blot.Conclusion The VEGF165 is expressed in the process of fracture healing with the highest expression period being during 72?h to 3 weeks after fracture.