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肝细胞生长因子促进骨形态发生蛋白9诱导小鼠骨髓间充质干细胞C3H10成骨分化的研究

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Keywords: 骨形态发生蛋白,肝细胞生长因子,骨髓间充质干细胞,成骨分化

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Abstract:

目的探讨肝细胞生长因子(hepatocytegrowthfactor,HGF)联合骨形态发生蛋白9(bonemorphogeneticprotein9,BMP9)诱导小鼠骨髓间充质干细胞(C3H10)成骨分化的影响,并探讨其可能的分子机制。方法用重组腺病毒介导BMP9在C3H10细胞中过表达,同时联用20ng/mL肝细胞生长因子HGF诱导C3H10细胞,Real-timePCR检测C3H10成骨分化相关基因骨钙蛋白(osteocalcin,OC)及骨桥蛋白(osteopontin,OPN)基因mRNA水平的表达变化;Westernblot检测OC、OPN的蛋白表达水平;ALP染色及ALP读数分析C3H10细胞碱性磷酸酶(alkalinephosphatase,ALP)活性的变化;茜素红S染色检测细胞的钙盐沉积;Real-timePCR检测经典信号通路Smad1/5/8在此诱导过程中的表达水平。?结果?BMP9单独作用能诱导C3H10细胞向成骨方向分化,并增强细胞ALP活性,单独HGF有较弱的成骨诱导作用,但与BMP9联用可显著增强C3H10细胞成骨特异性标志物OC与OPN的mRNA及蛋白表达水平(P<0.05),促进ALP活性,及晚期的钙盐沉积;HGF能刺激Smad1/5/8信号通路mRNA表达水平显著增高(P<0.05)。结论肝细胞生长因子可显著增强BMP9诱导C3H10细胞成骨分化的作用,可能是通过激活Smad经典信号通路来实现的。

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