Lu Yuezhi, Zhao Qian, Liu Yang, Zhang Ling, Li Danxue, Zhu Zhuoli, Gan Xueqi, Yu Haiyang
State Key Laboratory of Oral Diseases, West China School of Stomatology, Sichuan University, Chengdu, China; Department of Prosthodontics, West China School of Stomatology, Sichuan University, Chengdu, China.
State Key Laboratory of Oral Diseases, West China School of Stomatology, Sichuan University, Chengdu, China.
J Biomech. 2018 Apr 11;71:67-75. doi: 10.1016/j.jbiomech.2018.01.039. Epub 2018 Feb 8.
Low magnitude high frequency vibration (LMHFV) exhibits effectively anabolic effects on the bone tissue, and can promote osteogenic differentiation of mesenchymal stem cells (MSCs) in vitro. The role of p38 MAPK signaling in LMHFV-induced osteogenesis remains unclear. In this current study, LMHFV loading was applied to BMSCs in vitro, and cell proliferation, alkaline phosphatase (ALP), matrix mineralization, as well as osteogenic genes expression were assayed. The mechanism of mechanical signal transduction was analysed using PCR array, qRT-PCR and Western blot. LMHFV increased cell proliferation in the growth medium, while inhibited proliferation in the osteogenic medium. ALP activity, matrix mineralization and osteogenic genes expression of Runx2, Col-I, ALP, OPN and OC were increased by LMHFV. p38 and MKK6 genes expression, and p38 phosphorylation were promoted in LMHFV-induced osteogenesis. Inhibition of p38 MAPK with SB203580 and targeted p38 siRNA blunted the increased ALP activity and osteogenic genes expression by LMHFV. These findings suggest that LMHFV promotes osteogenic differentiation of BMSCs, and p38 MAPK signaling shows an important function in LMHFV-induced osteogenesis.
低强度高频振动(LMHFV)对骨组织具有有效的合成代谢作用,并能在体外促进间充质干细胞(MSCs)的成骨分化。p38丝裂原活化蛋白激酶(MAPK)信号通路在LMHFV诱导的成骨过程中的作用尚不清楚。在本研究中,将LMHFV加载应用于体外培养的骨髓间充质干细胞(BMSCs),检测细胞增殖、碱性磷酸酶(ALP)、基质矿化以及成骨基因表达情况。采用PCR阵列、定量逆转录聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法分析机械信号转导机制。LMHFV在生长培养基中增加细胞增殖,而在成骨培养基中抑制增殖。LMHFV增加了ALP活性、基质矿化以及Runx2、I型胶原(Col-I)、ALP、骨桥蛋白(OPN)和骨钙素(OC)的成骨基因表达。在LMHFV诱导的成骨过程中,p38和MKK6基因表达以及p38磷酸化均得到促进。用SB203580抑制p38 MAPK和靶向p38的小干扰RNA(siRNA)可减弱LMHFV诱导的ALP活性增加和成骨基因表达。这些结果表明,LMHFV促进BMSCs的成骨分化,并且p38 MAPK信号通路在LMHFV诱导的成骨过程中发挥重要作用。