国际口腔医学杂志 ›› 2017, Vol. 44 ›› Issue (6): 679-685.doi: 10.7518/gjkq.2017.06.011
薛令法1, 张岱尊2, 肖文林1, 于保军1
Xue Lingfa1, Zhang Daizun2, Xiao Wenlin1, Yu Baojun1
摘要: 目的 旨在研究p38分裂原激活的蛋白激酶(MAPK)信号通路和转录因子Osterix在间断性机械牵张力刺激下促进小鼠骨髓间充质干细胞(BMSC)成骨向分化的作用机制。方法 将C57BL/6J小鼠BMSC分为空白对照组、牵张力组和牵张力阻断组(p38MAPK通路抑制剂SB203580+牵张力),采用Flexercell体外细胞力学加载装置,施加频率为0.5 Hz、形变率为0.8%的牵张力,每天2次,每次30 min,分别在实验第1、3、5 d收获BMSC。实时荧光定量聚合酶链反应(RT-PCR)检测成骨基因ALP、COL I、OCN的mRNA水平变化情况,Western blot检测P-p38-MAPK蛋白的表达情况。通过小干扰核糖核酸(siRNA)技术沉默小鼠BMSC的osterix基因,Western blot检测Osterix 蛋白的表达情况,RT-PCR检测成骨相关基因ALP、COL I、OCN的mRNA水平变化情况。结果 牵张力作用后,可观察到成骨相关基因ALP、COL I、OCN及转录因子Osterix的mRNA水平增高。沉默osterix后,小鼠BMSC成骨相关基因ALP、COL I、OCN的mRNA水平也随之降低。Western blot结果显示,牵张力组小鼠BMSC中Osterix和P- p38-MAPK的蛋白质水平明显高于对照组(P<0.05)。SB203580作用后,小鼠BMSC成骨相关基因ALP、COL I、OCN和osterix的mRNA水平降低。结论 间断性牵张力可通过活化p38MAPK-Osterix通路促进小鼠BMSC成骨分化。
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