低氧诱导因子-1α在长骨发育中的表达研究
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作者Author单位AddressE-Mail
朱勋兵 ZHU Xun-bing 蚌埠医学院第一附属医院骨科组织移植安徽省重点实验室,安徽 蚌埠 233003 Shanghai Institute of Traumatology and Orthopedics,Shanghai 200025,China lfdeng@msn.com 
周建生 ZHOU Jian-sheng 蚌埠医学院第一附属医院骨科组织移植安徽省重点实验室,安徽 蚌埠 233003 Shanghai Institute of Traumatology and Orthopedics,Shanghai 200025,China  
肖玉周 XIAO Yu-zhou 蚌埠医学院第一附属医院骨科组织移植安徽省重点实验室,安徽 蚌埠 233003 Shanghai Institute of Traumatology and Orthopedics,Shanghai 200025,China  
邓廉夫 DENG Lian-fu 上海市伤骨科研究所  
期刊信息:《中国骨伤》2009年,第22卷,第8期,第599-601页
DOI:doi:10.3969/j.issn.1003-0034.yyyy.nn.zzz
基金项目:安徽省自然科学基金资助(编号:070413097)
中文摘要:

目的:通过动态观察小鼠长骨发育的演变过程,检测各发育时段中低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)表达及分布状况,探讨HIF-1α在小鼠长骨发育过程中的时空表达特点与作用。

方法:分别剖腹取得胎龄12.5、13.5、14.5、15.5、16.5、17.5 d的C57BL6小鼠的胎鼠,解剖显微镜下观察肢芽长骨的软骨形成及其软骨内骨化的动态形态演变过程。通过免疫组织化学方法观察HIF-1α蛋白在初级骨化中心形成中的表达与分布,进一步应用RT-PCR检测HIF-1α mRNA在胎鼠长骨各发育时段的表达状况及与骨标记基因骨钙素(OC)表达的关系。

结果:胚胎第13.5 d(E13.5)时,软骨性肢体形成,关节轮廓出现;E14.5时,软骨性长骨内形成小的骨化核,初级骨化中心刚刚出现,显现不透光骨化影,其后逐渐增大并向头尾两端延伸。免疫组化检测到E14.5时初级骨化中心内的软骨细胞核内见大量HIF 1α阳性黄色颗粒,之后很快消失。HIF-1α mRNA自E13.5至E15.5均大量表达,其后很快下降,维持低水平表达。

结论:长骨发育表现为软骨内成骨过程,存在低氧环境,伴随HIF-1α mRNA的规律性表达,这可能与长骨软骨内骨化过程的启动有关。
【关键词】低氧诱导因子  骨生成  小鼠
 
Expression of hypoxia inducible factor-1α in long bone development
ABSTRACT  

Objective: To observe dynamically the development of fetal long bone and detect the expression and distribution of HIF-1α,to investigate the expression pattern and possible effects of hypoxia inducible factor-1α (HIF-1α) in fetal long bone development of mouse.

Methods: E12.5,E13.5,E14.5,E15.5,E16.5 and E17.5 pregnant C57BL6 mice were sacrificed. After sacrifice,the embryos were delivered by caesarean section. The development of fetal long bone was dynamically observed by stereoscopic microscope,and the distributional expression of HIF-1α protein was detected by using method of immunohistochemistry. The expression of HIF-1α mRNA and osteoblast marker gene at various stage were also detected by using methods of reverse transcription-polymerase chain reaction(RT-PCR).

Results: The cartilaginous long bone began to form and joints outline arised at E13.5,then the primary ossification center was observed at E14.5,showing opaque ossification under stereoscopic microscope,and then the osteogenesis expanded and extended to both sides. Immunohistochemistry demonstrated lots of HIF-1α protein positive chondrcytes in the center of primary ossification at E14.5,then they decreased dramatically. HIF-1α mRNA expressed at high level from E13.5 to E15.5,and then decreased to low level.

Conclusion: Fetal long bone development pattern appeared to be endochondral osteogenisis process,existing hypoxia microenviroment may increase HIF-1α mRNA expression and thus initiate the cascade of endochondral osteogenisis.
KEY WORDS  Hypoxia inducible factor,HIF  Osteogenesis  Mice
 
引用本文,请按以下格式著录参考文献:
中文格式:朱勋兵,周建生,肖玉周,邓廉夫.低氧诱导因子-1α在长骨发育中的表达研究[J].中国骨伤,2009,22(8):599~601
英文格式:ZHU Xun-bing,ZHOU Jian-sheng,XIAO Yu-zhou,DENG Lian-fu.Expression of hypoxia inducible factor-1α in long bone development[J].zhongguo gu shang / China J Orthop Trauma ,2009,22(8):599~601
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