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p38 Kinase Participated in BMP9-induced Osteogenic Differentiation of C3H10T1/2 Mesenchynal Stem Cells |
XU Dao-jing, WANG Jin, HE Juan-wen, HU Jing, WENG Ya-guang, LUO Jin-yong |
Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education,Chongqing Medical University,Chongqing 400016,China |
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Abstract Objective:Analysis the functional role of p38 kinase in BMP-9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells. Methods:C3H10T1/2 cells were infected by recombinant adenovirus expressing BMP9, then the total protein level and phosphorylated form of p38 kinase were determined by Western blot. After treatment C3H10T1/2 cells with p38 specific inhibitor SB203580 or using RNA interference to silence expression of p38, the early osteogenic marker ALP activity was detected by quantitative and staining assay, later osteogenic marker calcium deposition was determined by Alizarin Red S staining, expression level of Smad6 and Smad7 was analyzed by Real time PCR. Animal assay was carried out to confirm that whether p38 silence can result in inhibition of entopic bone formation induced by BMP9 in vivo. Results: BMP9 did not change total protein level of p38, however, BMP9 increased the phosphorylated form of p38 kinase. P38 specific inhibitor SB203580 dose-dependently decreased ALP activity induced by BMP9 of C3H10T1/2 cells. Gene silence of p38 by RNA interference also led to a reduction of ALP activity. Furthermore, SB203580 markedly inhibited calcium deposition, reduced BMP9-induced expressions of Smad6 and Smad7. Moreover, p38 gene silence was also showed to inhibit entopic bone formation induced by BMP9 in vivo. Conclusion: The p38 kianse may invovlve in BMP9 induced osteoblast commitment of C3H10T1/2 mesenchymal stem cells.
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Received: 23 December 2010
Published: 27 May 2011
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