HIF-1α在软骨内骨化过程中的表达研究

Study on the Expression of HIF-1α in Entochondrostosis

目的通过检测在不同日龄大鼠股骨中HIF-1α和VEGF的表达情况和它们之间的关系,探讨HIF-1α在软骨发育中血管侵入中的作用。方法选取出生后1、3、5、7、9、11、13、15天的大鼠,每组6只,大鼠断头术处死后取双侧股骨,常规固定,包埋,切片,采用HE染色法观察血管侵入的情况。采用免疫组织化学方法观察HIF-1α、VEGF蛋白的表达情况,RT-PCR法检测HIF-1α,VEGF mRNA的表达情况。并用SPSS 16.0软件进行数据分析。结果第1、3、5天的大鼠股骨内只有少量血管侵入,第7、9、11天的大鼠股骨内出现大量的血管侵入,第13,15天的大鼠股骨内的软骨开始钙化血管减少。HIF-1α和VEGF在第1、3、5天的大鼠股骨内弱表达,在第7、9、11天的大鼠股骨内强表达,在第13、15天的大鼠股骨内弱表达。结论在骨发育的过程中,HIF-1α可能通过激活其下游基因表达VEGF诱导外源性血管的侵入。

Objective To investigate the expression of HIF-1α in entochondrostosis,and to study it＇s role in the process of vascular intrusion.Methods The morphology change in rat＇s femoral bone development were observed using HE staining method.The protein expression of HIF-lα and VEGF in femoral bone was detected by immunohistochemistry,and the mRNA expression of HIF-lα and VEGF was detected by reverse transcription-polymerase chain reaction（RT-PCR） method.Results Many cartilage cells were seen in femoral bone at the 1st day after the birth;many vessels began to invade at the 7th day;at the 13th day,the cartilage matrix began to calcification,vessels and cartilage cells were reduced.The protein expression of HIF-lα and VEGF was increased at the 7th day,while decreased at the 13th day.The mRNA expression of HIF-lα and VEGF went up to the maximum at 7th day the time,and then decreased.Conclusion During femoral bone development,hypoxia may induce HIF-1 expression,then activates it＇s down stream gene VEGF expression,which would induce the intrusion of exogenous vascular,promote endochondral osteogenesis.