摘要
目的研究丝裂原激活的蛋白激酶（mitogen-activated protein kinases,MAPKs）的亚型p38 MAPK在植物雌激素金雀异黄酮（Genistein）促进小鼠骨髓间充质干细胞（bone marrow-derived mesenchymal stem cells ,BMSCs）向成骨细胞分化过程中的作用.方法 BMSCs用无酚红α-MEM（含经活性碳吸附的10% FBS、β-磷酸甘油、维生素C）培养,先用Genistein处理细胞,观察BMSCs向成骨细胞分化情况,然后用SB203580（p38 MAPK通路阻断剂）以及PD98059（p44/42 MAPK通路阻断剂）阻断相应的通路后,再用Genistein处理细胞,观察BMSCs向成骨细胞分化情况,并同时观察上述处理后MAPK通路的变化.测定碱性磷酸酶（ALP）活性和钙（Ca）沉积量反映BMSCs向成骨细胞分化状况,用Western blot来检测MAPK通路是否激活.结果 Genistein（0.01,0.1,1 μmol·L-1）剂量依赖性增加小鼠BMSCs细胞内ALP活性和细胞外Ca沉积量,并同时引起p38MAPK通路的激活和p44/42MAPK通路的抑制.SB203580预处理能减弱Genistein刺激引起的p38MAPK通路的激活并同时阻止Genistein诱导的BMSCs向成骨细胞分化.结论 Genistein在0.01～1 μmol·L^-1剂量范围内可通过p38MAPK通路促进小鼠BMSCs向成骨细胞分化.
Aim To investigate the role of p38 mitogen-activated protein kinases （MAPKs） in genisteininduced osteoblastic differentiation of mouse bone marrow-derived mesenchymal stem cells （BMSCs）. Methods Mouse BMSCs＇ cultured in phenol red-free α- MEM containing 10% V/V FBS, were added β-glycerophosphate and ascorbic acid for inducing osteoblastic differentiation, and treated with 0. 01 - 1 μmol . L^-1 genistein and/or SB203580 and PD98059. The temporal sequence of osteoblastie differentiation in BMSCs cultures was assayed by measuring alkaline phosphatase activity （ALP） and calcium deposition. The MAPK phosphorylation level was detected by Westernblot. Results Genistein （0.01 -1 μmol . L^-1） showed a dose-dependent effect on osteoblastie differentiation as evidenced by increased alkaline phosphatase （ALP） activity and calcium deposition in mouse BMSCs cultures. Genistein （ 1 μmol.L^-1 ）_induced osteoblastic differentiation of BMSCs was diminished by p38 MAPK inhibitor but not the p44/42 MAPK inhibitor. The effects of Genistein were associated with rapid and sustained activation of p38 MAPK in the BMSCs cultures, which was blocked by SB203580 （1 μmol.L^-1）. In contrast, Genistein treatment resulted in inactivation of p42/44MAPK, which was further attenuated by PD98059（25 μmol . L^-1 ）. Conclusion p38 MAPK plays an important role in genistein-induced osteoblastic differentiation of mouse BMSCs cultures.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2006年第6期683-687,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30171085)
教育部高等学校优秀青年教师教学科研奖励计划资助项目(TRAPOYT
No200026)
关键词
金雀异黄酮
骨髓间充质干细胞
成骨细胞分化
丝裂原激活的蛋白激酶
Genistein
bone marrow-derived mesen chymal stem cells （BMSCs）
osteoblastic differentia tion
mitogen-activated protein kinases （MAPKs）
