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《Journal of Clinical Rehabilitative Tissue Engineering Research》 2009-37
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Effects of p38MAPK on the proliferation, differentiation and apoptosis of murine osteoblasts

Wang Xiao-hui1, Tang Xu-lei21Department of Endocrine, Gansu Hospital of Traditional Chinese Medicine, Lanzhou 730000, Gansu Province, China; 2Department of Endocrine, the First Hospital of Lanzhou University, Lanzhou 730000, Gansu Province, China  
BACKGROUND: p38MAPK is one of signal transduction pathway of MAPK. However, there are no reports addressing how p38MAPK control the proliferation and differentiation of osteoblasts, as well as if it is concerned with the apoptosis of osteoblasts. OBJECTIVE: To investigate the effects of p38MAPK on the proliferation, differentiation and apoptosis of murine osteoblasts. DESIGN, TIME AND SETTING: In vitro single sample controlled observation at a cytology level was performed in the Cell Culture Room, Orthopaedic Institute, General Hospital of Lanzhou Military Area Command of Chinese PLA between May 2007 and March 2008. MATERIALS: Nine newborn SD rats within 24 hours were used to isolate osteoblasts; SB203580 (the inhibitor of p38MAPK) was offered by Sigma. METHODS: Different doses of 17β-Estradiol and Puerarin (1×10-6, 1×10-7, 1×10-8 mol/L) were added into the medium of the first generation osteoblasts harvested from the skull of newborn SD rats, serving as drug stimulus groups; in the inhibitor group, 10 μmol/L SB203580 was added 30 minutes in advance to block signal transduction pathway, followed by drug stimulation; Blank control group was also set. MAIN OUTCOME MEASURES: The proliferation and alkaline phosphatase activity of cells in different groups were determined with methyl thiazolyl tetrazolium method and p-Nitrophenyl phosphate method. The apoptosis rate was analyzed by flow cytometry with Annexin V-FITC/PI kit. RESULTS: 17β-Estradiol or Puerarin apparently stimulated the proliferation and differentiation of murine osteoblasts, with significant differences compared with blank control group (P 0.05); After p38MAPK signal transduction pathway was blocked, cell proliferation was not inhibited (P 0.05); the differentiation was apparently inhibited, with extremely significant differences compared with non-blocked groups (P 0.01). Flow cytometry results showed that, drug stimulus could reduce the apoptotic rate of osteoblasts apparently compared with control group (P 0.05); SB203580 had no apparent influence the apoptosis (P 0.05). CONCLUSION: 17β-Estradiol and Puerarin could promote the proliferation and differentiation of murine osteoblasts, as well as inhibits the apoptosis; p38MAPK plays an important role in the differentiation of osteoblasts, but it has no significant effect on the proliferation and the apoptosis.
【CateGory Index】: R363
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