| Objective To investigate the effects of p38MAPK on the proliferation,differentiation and inhibited apoptosis of murine osteoblasts by 17B-estradiol,puerain and resveratrol.Methods 1.The newborn SD mice calvarium osteoblasts(OB)were isolated and digested with trypsin and collagenase repeatedly,which were then cultured and observed the morphologic character both in the light and phase-contrast microscopy. OB was identified by cytochemical staining of alkaline phosphatase(ALP)and the mineralization nodes were stained by different staining.2.The second passagae of OB were place in 96-well culture plates and incubated for 24h,divided to control,various concentrations of 17β-estradiol,puerarin and resveratrol group(10-8-10-6mol·L-1),and 17β-estradiol,puerarin and resveratrol were added into SB203580(10μmol·L-1)group, the inhibitor of p38MAPK,were added pretime.The proliferation,differentiation,mainerlization and calcium deposition of different groups were observed.3.After 8d using RT-PCR assay to determine collagen I mRNA expression,and divided to the control,17β-estradiol,puerarin and resveratrol group(10-8mol·L-1),blocking group.4. After 24h the second passagae of OB being no serum incubating and divided to control, 17β-estradiol,puerarin and resveratrol group(10-8mol·L-1),blocking group,after 72h and apoptosis rate was analyzed by FCM with Annexin V-FITC/PI kit.Results 1.17β-Estradiol,puerarin and resveratrol at the concentration of 10-8-10-6mol·L-1had apparently stimulated the proliferation of murine osteoblast(P＜0.05,P＜0.01,vs control),The blocking group had increased the value of MTT(P＜0.05, P＜0.01,vs control),The contain and the no-contain blocking group have closed to each other.2.17β-Estradiol,puerarin and resveratrol at the concentration of 10-8-10-6mol·L-1 had apparently stimulated the differentiation of murine osteoblast(P＜0.05,P＜0.01,vs control),The blocking group had decreased the ALP activity(P＜0.05,P＜0.01,vs control).3.17β-Estradiol,puerarin and resveratrol at the concentration of 10-8-10-6mol·L-1had apparently stimulated the mineral nodes and the calcium deposition of murine osteoblast(P＜0.05,P＜0.01,vs control),The blocking group had apparently decreased the mineral nodes and the calcium deposition value(P＜0.01,vs control).4. 17β-Estradiol,puerarin and resveratrol at the concentration of 10-8mol·L-1had apparently stimulated the collagen I mRNA expression of murine osteoblast(P＜0.05, vs control),the blocking group had apparently blocked the collagen I mRNA expression (P＜0.01,vs control).5.17β-Estradiol,puerarin and resveratrol at the concentration of 10-8mol·L-1had inhibited the apoptosis of osteoblasts apparently(P＜0.05,vs control), the blocking group had apparently inhibited the apoptosis of osteoblasts(P＜0.05,vs control),The contain and the no-contain blocking group have not significant differented.Conclusion 17β-Estradiol,puerarin and resveratrol apparently stimulated the proliferation,differentiation,mineral nodes,calcium deposition,the collagen I mRNA expression and inhibited the apoptosis of murine osteoblast.P38MAPK signal transduction pathway plays an important role in the differentiation,mineral nodes, calcium deposition and the collagen I mRNA expression of murine osteoblast,but it has significant effect on the proliferation and inhibited the apoptosis of murine osteoblast. |
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