Dkk1对骨形态发生蛋白9诱导间充质干细胞成骨分化的影响  

Effects of Dkk1 on bone morphogenetic protein 9 induced mesenchymal stem cells osteogenic differentiation

作　　者：廖军义 林良波[1] 周年[1] 梁熙[1] 胡宁[1] 毕杨[2] 黄伟[1] 

机构地区：[1]重庆医科大学附属第一医院骨科,重庆400016 [2]重庆医科大学附属儿童医院干细胞实验室,重庆400014

出　　处：《重庆医科大学学报》2014年第5期578-582,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:31070875)

摘　　要：目的:探讨经典Wnt信号通路胞外拮抗因子Dkk1对骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)诱导间充质干细胞(mesenchymal stem cells,MSCs)成骨分化的影响。方法:以C3H10T1/2为目的细胞,通过重组腺病毒介导BMP9过表达,联用重组腺病毒Dkk1抑制经典Wnt信号通路。检测各处理组碱性磷酸酶(alkaline phosphatase,ALP)活性变化、钙盐沉积、骨钙素(osteocalcin,OC)、骨桥素(osteopontin,OPN)变化;体内异位成骨实验分析Dkk1对BMP9诱导MSCs成骨分化的影响。结果:BMP9可诱导C3H10T1/2细胞成骨分化;Dkk1显著降低BMP9诱导的C3H10T1/2细胞ALP活性(F=1 467.21,P=0.000)、钙盐沉积、OC(F=32.95,P=0.000)和OPN(F=127.23,P=0.000)蛋白表达;异位成骨模型中Dkk1抑制BMP9诱导的C3H10T1/2成骨分化和骨块成熟。结论:Wnt信号通路胞外拮抗因子Dkk1可降低BMP9诱导的C3H10T1/2成骨分化,BMP9诱导的C3H10T1/2细胞成骨分化可能需要经典的Wnt信号通路参与。Objective：To investigate the effect of canonical Wnt signaling antagonist Dickkopf1（Dkk1）on bone morphogenetic protein 9（BMP9）induced C3H10T1/2 mesenchymal stem cells（MSCs）osteogenic differentiation. Methods：C3H10T1/2 cells were infected by recombinant adenovirus expressing green fluorescent protein（GFP）,BMP9 and/or Dkk1. Early osteogenic marker,alkaline phosphatase（ALP）activity was detected by ALP activity assay and staining. Later osteogenic marker calcium deposition was determined by Alizarin Red S staining. Western blot was used to detect the middle osteogenic marker osteopontin（OPN）and osteocalcin（OC）at protein level. Ectopic bone formation assay was carried out to make sure the influence of Dkk1 on BMP9 induced osteogenic differentiation of mesenchymal stems cells in vivo. Results：Dkk1 effectively inhibited BMP9 induced ALP activity,calcium deposition,OPN and OC protein expression in C3H10T1/2 cells. Athymic nude mice subcutaneous injection C3H10T1/2 cells showed that Dkk1 inhibited BMP9 induced ectopic bone formation and calcification degree in vivo. Conclusions：Wnt signaling antagonist Dkk1 can dramatically inhibit BMP9 induced C3H10T1/2 cells osteogenic differentiation and BMP9 induced osteogenic differentiation may require functional canonical Wnt signaling.

关 键 词：骨形态发生蛋白9 经典Wnt信号通路 DKK1 间充质干细胞 骨向分化 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]