低氧诱导因子-1α和VHL对小鼠软骨内成骨过程的调控机制  

Role of hypoxia inducible factor-1α and VHL in murine endochondral ossification

作　　者：邵进[1] 邓廉夫[1] 齐进[1] 周琦[1] 王君[1] 魏立[1] 王晋申[1] 钱念东[1] 

机构地区：[1]上海交通大学医学院瑞金医院骨科上海市伤骨科研究所,上海200025

出　　处：《上海交通大学学报（医学版）》2009年第11期1305-1310,共6页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(30672145);上海市科委2006"登山计划"(06DZ22020);上海市科委2007"创新行动"(07DZ22008)~~

摘　　要：目的研究低氧诱导因子1α(HIF-1α)和von Hippel-Lindau(VHL)在成骨细胞水平对小鼠软骨内成骨过程的调控机制。方法以HIF-1α或VHL基因条件性敲除小鼠为实验对象,分别于4、8、12周龄时,采用组织化学染色观察、显微CT扫描、骨小梁面积测量、钙元素含量检测、四环素荧光双标记观察、Real-time PCR及Western blotting等方法,观察和比较基因敲除小鼠与野生型小鼠(对照组)软骨内成骨过程的差异。结果与野生型对照小鼠比较,HIF-1α基因敲除组小鼠在软骨内成骨过程中血管内皮生长因子(VEGF)表达减少,新骨形成速度减慢,钙元素含量和骨小梁面积减少(P<0.05);而VHL基因敲除组小鼠在软骨内成骨过程中VEGF表达增加,新骨形成速度加快,钙元素含量和骨小梁面积增加(P<0.001)。结论在小鼠软骨内化骨过程中,VHL/HIF-1α信号通路对VEGF表达具有调控作用,通过调节血管形成,最终影响骨量形成。Objective To investigate the role of hypoxia inducible factor-1α （HIF-1α） and von Hippel-Lindau （VHL） in murine endochondral ossification. Methods The knockout of HIF-1α or VHL gene in murine osteoblasts was accomplished by conditional knockout technique at 4th,8th and 12th week,and the differences between wild-type group and knock-out group in endochondral ossification were detected by HE staining,micro-CT scanning,trabecular bone area measurement,calcium content measurement,tetracycline fluorescence labeling,Real-time PCR and Western blotting.Results After knockout of HIF-1α gene in osteoblasts,the expression of vascular endothelial growth factor（VEGF） reduced,the rate of new bone formation stepped down,the content of calcium became less,and the trabecular bone volume decreased （ P 0.05）. After knockout of VHL gene in osteoblasts,the expression of VEGF increased,the rate of new bone formation stepped up,the content of calcium became more,and the trabecular bone volume was promoted （ P 0.001）.Conclusion During murine endochondral ossification,VHL/HIF-1α signal pathway promotes angiogenesis through the stimulation of VEGF expression,which subsequently accelerates osteogenesis.

关 键 词：低氧诱导因子-1Α von HIPPEL-LINDAU 软骨内成骨 

分 类 号：R681.3[医药卫生—骨科学] Q78[医药卫生—外科学]