摘要
目的 探讨诱导条件下成纤维细胞表达成骨细胞特异性转录子 -核结合因子 (Cbfa1)的可能性及成纤维细胞表达成骨表型的发生机理。 方法 分离、纯化人皮肤成纤维细胞 ,使其生长在分别含一定浓度肿坏死因子 (TNF) α、骨诱导蛋白 (BMP) 2、TNF α加BMP 2的条件培养液中 ,采用逆转录聚合酶链反应和免疫组织化学技术 ,分别检测Cbfa1、骨钙素、H ras、BMPⅠ型受体 (BMPR ⅠA)和Ⅰ、Ⅲ型胶原mRNA表达及Ras、BMPR ⅠA蛋白形成状况。 结果 TNF α可诱导成纤维细胞由表达Ⅰ、Ⅲ型胶原表型向表达Ⅰ型胶原转化 ;促进成纤维细胞Ras的形成 ,在TNF α作用 1、3h和 1周后 ,可使成纤维细胞H rasmRNA的相对表达量分别升高 39 5 %、7 4%和 43 2 % ;TNF α作用 1h后的成纤维细胞可产生BMPR ⅠA。联合应用TNF α与BMP 2可诱导成纤维细胞表达Cbfa1、骨钙素mRNA。 结论 TNF α与BMP 2联合应用 ,可刺激成纤维细胞表型转化 。
Objective To investigate the probabilities of core-biding factor a1 (Cbfa1) expression by human skin fibroblasts induced in vitro. Methods The fibroblasts were isolated, purified from human skin, and were grown in incubation in the media of TNF-α, BMP-2, and combined TNF-α and BMP-2 at certain concentrations, respectively. The changes in biological features of these fibroblasts correlated with osteogenesis were detected by immunohitochemistry and RT-PCR assay. Results TNF-α could switch phenotype of collagen in fibroblasts from TypeⅠand Ⅲ to TypeⅠand induce fibroblasts to express Ras and BMP typeⅠreceptor (BMPR-ⅠA). TNF-α in combination with BMP-2 could induce fibroblasts to express Cbfa1 and osteocalcin mRNA. Conclusion Human skin fibroblast could be induced into pro-osteoblast expressing Cbfa1, an osteoblast-specific transcription factor and a regulation of osteoblast differentiation, and combined use of TNF-αand BMP-2 was one of the regulating factors.
出处
《中华外科杂志》
CAS
CSCD
北大核心
2002年第8期592-595,共4页
Chinese Journal of Surgery
基金
国家自然科学基金 ( 30 0 70 75 9)
上海市青年科技启明星培养计划资助项目
