骨形态发生蛋白9促进小鼠颅骨间充质祖细胞成骨分化的实验研究  

BMP9 effectively induces osteogenic differentiation of immortalized calvarial mesenchymal progenitor cells

作　　者：宿旭东 邓锋[1,2] 汤寅虹 周鹏飞[1,2] 梁盼盼[1,2] 蓝丹[1,2] 黄恩毅[1,2] SU Xu-dong DENG Feng TANG Yin-hong ZHOU Peng-fei LIANG Pan-pan LAN Dan HUANG En-yi

机构地区：[1]重庆医科大学附属口腔医院,口腔疾病与生物医学重庆市重点实验室,重庆401147 [2]重庆医科大学口腔医学院,重庆市高校市级口腔生物医学工程重点实验室,重庆401147

出　　处：《军事医学》2016年第12期984-987,993,共5页Military Medical Sciences

基　　金：国家自然科学基金资助项目(81301551);重庆市基础科学与前沿技术研究专项资助项目(cstc2013jcyj A10022);重庆高校创新团队建设计划资助项目(KJTD201304)

摘　　要：目的研究腺病毒载体介导的骨形态发生蛋白9(BMP-9)对小鼠永生化颅骨间充质祖细胞(i CAL)成骨分化的诱导作用。方法以i CAL为模型,设置绿色荧光蛋白(GFP)对照组和BMP-9诱导组,于诱导后的3、5、7 d对早期成骨分化指标碱性磷酸酶(ALP)活性进行定性和定量检测;于诱导后的14 d通过茜素红染色检测骨钙结节形成;荧光定量PCR法和Western印迹检测成骨细胞分化标志基因Runx2、OCN表达情况。结果 BMP-9诱导组细胞内ALP活性高于GFP对照组,差异具有统计学意义(P<0.05);BMP-9诱导组形成较多的钙盐结节沉积;荧光定量PCR检测结果显示诱导组中Runx2、OCN的mRNA表达量明显上调,差异具有统计学意义(P<0.05),Western印迹检测3 d Runx2蛋白表达上调,5 d OCN蛋白表达上调。结论 BMP-9促进小鼠永生化i CAL成骨分化。Objective To investigate the effect of adenovirns-bone morphogenic protein 9 （Ad-BMP9） on osteogenic differentiation of immortalized calvarial mesenchymal progenitor cells （iCALs）. Methods iCALs were infected with adenoviral vectors encoding BMP-9 or green fluorescent protein（GFP） and the early osteogenic differentiation was assessed by detecting alkaline phosphatase（ALP） activity after being cultured for 3, 5 and 7 days. 14 days after infection, alizarin red S staining was performed to study the formation of osteogenic calcium nodules. The expression of osteogenic marker genes Runx2 and OCN was assessed by quantitative real-time （RT）-PCR and Western blotting.. Results Significant increases in ALP activity and in the expressions of Runx2 and OCN were detected in BMP-9 treated iCALs compared with GFP-treated cells（ P 〈0.05 ）. Meanwhile, alizarin red S staining showed that more mineralized nodules were found in the BMP-9 induced group. Conclusion BMP-9 can promote the osteogenic differentiation of iCALs.

关 键 词：骨形态发生蛋白质类 小鼠 颅骨间充质祖细胞 成骨分化 颅骨修复 碱性磷酸酶 

分 类 号：R782[医药卫生—口腔医学]