Shh促进BMP9诱导的小鼠间充质干细胞C3H10T1/2的成骨分化???

基础医学与临床 ›› 2014, Vol. 34 ›› Issue (12): 1629-1634.

Shh促进BMP9诱导的小鼠间充质干细胞C3H10T1/2的成骨分化???

李丽,董谦,冯巧灵,王玉凤,何通川,罗进勇

重庆医科大学

收稿日期:2014-05-06 修回日期:2014-06-20 出版日期:2014-12-05 发布日期:2014-11-25
通讯作者: 罗进勇 E-mail:luojinyong@sina.com
基金资助:
BMP9-MAPK信号途径调控间充质干细胞成骨分化;BMP9/PI3K/AKT信号途径调控间充质干细胞成骨分化;聚焦超声治疗肿瘤后坏死组织的转归及其效应研究

Shh enhances BMP9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells

Received:2014-05-06 Revised:2014-06-20 Online:2014-12-05 Published:2014-11-25

摘要/Abstract

摘要： 目的观察Shh蛋白对骨形态发生蛋白9（BMP9）诱导的小鼠间充质干细胞(MSCs)C3H10T1/2成骨分化的影响，并初步探讨其作用机制。方法 Shh腺病毒和BMP9作用于C3H10T1/2细胞，碱性磷酸酶（ALP）检测ALP变化，茜素红S染色检测钙盐沉积，RT-PCR检测Shh、BMP9、骨桥蛋白（OPN）、骨钙素（OCN）以及成骨相关基因Id1、Id2、Id3、CTGF和Runx2的表达，Western blot检测OPN、OCN、Runx2、Dlx5和p-Smad1/5/8的蛋白水平，荧光素酶报告基因检测Smad1/5/8的转录活性。结果 Shh不影响BMP9的表达，但可增强由BMP9诱导的C3H10T1/2细胞早晚期成骨分化（P<0.05），并促进BMP9诱导的成骨相关基因的表达（P<0.05）；Shh促进了BMP9诱导的Smad荧光素酶活性（P<0.05），但对其磷酸化并无影响。结论 Shh可促进BMP9诱导的小鼠C3H10T1/2细胞的成骨分化。

关键词: 关键词：间充质干细胞, 骨形态发生蛋白9, Shh, 成骨分化

Abstract: Objective To analysis the effect of Shh on BMP9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells and the mechanism involved. Methods C3H10T1/2 cells were treated with adenovirus Shh or/and BMP9, then the ALP activity was detected by quantitative and staining assay, calcium deposition was detected by Alizarin Red S staining, the expressions of Shh, BMP9, OPN, OCN, Id1, Id2, Id3, CTGF and Runx2 were detected by RT-PCR, the expressions of BMP9, OPN, OCN, Runx2, Dlx5 and p-Smad1/5/8 were detected by Western blot, Luciferae reporter assay was detected by quantitative. Results Shh can not influence the expression of BMP9 but increase the ealy and later osteogenic differentiation and the expression of osteogenesis related genes of C3H10T1/2 cells induced by BMP9. Furthermore, Shh also led to luciferase activity but not the phosphorylation activity of canonical Smad pathway stimulated by BMP9. Conclusion Shh promotes BMP9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells.

Key words: Key words: mesenchymal stem cells, BMP9, Shh, osteogenic differentiation

中图分类号:

Q254

李丽董谦冯巧灵王玉凤何通川罗进勇. Shh促进BMP9诱导的小鼠间充质干细胞C3H10T1/2的成骨分化???[J]. 基础医学与临床, 2014, 34(12): 1629-1634.

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