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目的探讨震荡流体剪切力(oscillatory shear stress,OSS)通过ERK5信号通路在诱导成骨细胞增殖中发挥的作用。方法对成骨MC3T3-E1细胞进行不同的处理,分为正常组、OSS组、XMD8-92组和OSS+XMD8-92组。采用MTT实验分别测定4组细胞的增殖活性并绘制生长曲线;蛋白免疫印迹法分别检测P-ERK5、ERK5和Cyclin D1等蛋白水平变化。结果 OSS可显著增加成骨MC3T3-E1细胞增殖活性,但此效应可被ERK5高选择性抑制剂XMD8-92阻断。OSS可显著上调Cyclin D1的表达,而XMD8-92可显著下调OSS诱导的Cyclin D1的表达。结论 OSS通过激活ERK5信号通路促进成骨细胞增殖,Cyclin D1是ERK5信号通路下游的重要靶点基因。
Objective To investigate the role of oscillatory shear stress (OSS) in inducing osteoblast proliferation through ERK5 signaling pathway. Methods The osteoblastic MC3T3-E1 cells were treated with different methods and divided into normal group, OSS group, XMD8-92 group and OSS + XMD8-92 group. MTT assay was used to determine the proliferation activity of four groups of cells and the growth curve was drawn. Western blotting was used to detect the protein levels of P-ERK5, ERK5 and Cyclin D1. Results OSS significantly increased the proliferation activity of osteoblastic MC3T3-E1 cells, but this effect was blocked by XMD8-92, a ERK5-selective inhibitor. OSS significantly up-regulated Cyclin D1 expression, while XMD8-92 significantly down-regulated OSS-induced Cyclin D1 expression. Conclusion OSS can promote the proliferation of osteoblasts by activating ERK5 signaling pathway. Cyclin D1 is an important target gene downstream of ERK5 signaling pathway.