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

发布时间：2018-01-18 07:03

本文关键词：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　出处：《第三军医大学学报》2014年23期 　论文类型：期刊论文

【摘要】：目的探讨肝细胞生长因子(hepatocyte growth factor,HGF)联合骨形态发生蛋白9(bone morphogenetic protein 9,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20 ng/m L肝细胞生长因子HGF诱导C3H10细胞,Real-time PCR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;Western blot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(alkaline phosphatase,ALP)活性的变化;茜素红S染色检测细胞的钙盐沉积;Real-time PCR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(P0.05),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(P0.05)。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。
[Abstract]:Objective to investigate hepatocyte growth factor. HGF combined with bone morphogenetic protein 9bone morphogenetic protein 9. BMP9) induced osteogenic differentiation of mouse bone marrow mesenchymal stem cells (BMP9). Methods Recombinant adenovirus mediated BMP9 overexpression in C3H10 cells. At the same time, C3H10 cells were induced by 20 ng/m L hepatocyte growth factor HGF. Osteocalcin (OC) and osteopontin (osteopontin) were detected by Real-time PCR. The change of mRNA expression level of OPN gene; Western blot was used to detect the protein expression of OCG-OPN. The activity of alkaline phosphatase (ALP) in C3H10 cells was analyzed by ALP staining and ALP reading. Calcium deposition was detected by alizarin red S staining. Real-time PCR was used to detect the expression level of classical signal pathway Smad1/5/8 during the induction. Results. BMP9 alone could induce C3H10 cells to differentiate into osteoblasts. The activity of ALP was enhanced and the osteogenesis induced by HGF alone was weaker. However, combined with BMP9 could significantly enhance the mRNA and protein expression level of OC and OPN in C3H10 cells and promote the activity of ALP. And late calcium deposition; HGF could stimulate the expression of mRNA in Smad1/5/8 signaling pathway and increase the level of mRNA expression significantly (P0.05). Conclusion Hepatocyte growth factor can significantly enhance the osteogenic differentiation of C3H 10 cells induced by BMP9. This may be achieved by activating the classical Smad signaling pathway.
【作者单位】：重庆医科大学附属儿童医院儿科研究所干细胞实验室儿童发育疾病研究教育部重点实验室儿科学重庆市重点实验室重庆市儿童发育重大疾病诊治与预防国际科技合作基地;
【基金】：国家自然科学基金青年科学基金(81100309)~~
【分类号】：R329
【正文快照】： 点实验室,儿科学重庆市重点实验室,重庆市儿童发育重大疾病诊治与预防国际科技合作基地)骨髓间充质干细胞(mesenchymal stem cell,MSCs)是一类具有自我更新能力和多向分化潜能的多能干细胞,作为骨组织工程中的种子细胞被广泛研究[1-2],然而在治疗骨科疾病中MSCs分化为成骨细胞

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