摘要
目的探讨血管内皮生长因子(VEGF)在鼠成骨细胞分化过程中的作用。方法利用鼠原代FRC细胞培养系统(第0、5、9、14天)和取材新生鼠颅盖骨组织(5例),采用反转录聚合酶链反应(RTPCR)和免疫组织化学染色方法,检测VEGF及其受体Flt1和KDR/Flk1在FRC细胞分化、矿化过程中mRNA和在新生鼠颅盖骨组织中蛋白质的表达。结果在控制培养条件下,FRC细胞出现明显的成骨特性。结合FRC细胞分化特性,RTPCR检测发现VEGF及其受体Flt1和KDR/Flk1在FRC细胞中从分化到矿化过程中,mRNA表达逐渐增强。特别是在矿化后第9天和第14天表达水平明显增强。免疫组织化学染色证实了RTPCR结果,新生鼠颅盖骨中的成骨细胞同时表达VEGF及其受体Flt1和KDR/Flk1。结论VEGF及其受体Flt1和KDR/Flk1在成骨细胞分化和矿化过程中表达逐渐增强。作为一种重要的血管性因子,VEGF的自分泌作用方式在成骨细胞的分化中起重要作用。
Objective To study the potential role of VEGF during rat osteoblast differentiation.Methods Rat primary FRC cell culture system (day 0,5,9,14) was produced and calvaria from 5 neonatal rats were collected.The mRNA expressions of VEGF and its receptors Flt 1 and KDR/Flk 1 in FRC cells during FRC differentiation and mineralization were examined by RT PCR and these protein expressions in calvaria were detected by immunohistochemistry.Results The FRC cell strongly expressed characteristics of the osteoblast phenotype in the control cultures.The mRNA expressions of VEGF and its receptors Flt 1 and KDR/Flk 1 gradually increased with the process from differentiation to mineralization in FRC cells,and the intensity was distinctly improved after day 9 and day 14 of mineralization.Immunolocalizations substantiated the results of RT PCR,VEGF together with its two receptors were co expressed in osteoblasts of neonatal rats calvaria.Conclusion The expressions of VEGF and its receptors Flt 1 and KDR/Flk 1 gradually increased in FRC cells during cell differentiation.Acting as an important factor of angiogenesis,VEGF might play an important autocrine function during osteoblast differentiation.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2004年第7期852-854,F003,共4页
Chinese Journal of Experimental Surgery
