CAJ | 학술논문

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.
목적 연구저양유도인자1α (HIF-1α)화von Hippel-Lindau (VHL)재성골세포수평대소서연골내성골과정적조공궤제.방법 이HIF-1α혹VHL기인조건성고제소서위실험대상,분별우4、8、12주령시,채용조직화학염색관찰、현미CT소묘、골소량면적측량、개원소함량검측、사배소형광쌍표기관찰、Real-time PCR급Western blotting등방법,관찰화비교기인고제소서여야생형소서(대조조)연골내성골과정적차이.결과 여야생형대조소서비교,HIF-1α기인고제조소서재연골내성골과정중혈관내피생장인자(VEGF)표체감소,신골형성속도감만,개원소함량화골소량면적감소(P<0.05);이VHL기인고제조소서재연골내성골과정중VEGF표체증가,신골형성속도가쾌,개원소함량화골소량면적증가(P<0.001).결론 재소서연골내화골과정중,VHL/HIF-1α신호통로대VEGF표체구유조공작용,통과조절혈관형성,최종영향골량형성.
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.