周期性和持续性流体剪切力对成骨细胞OPG、RANKL蛋白表达的影响  

The Effect of Cyclic and Continuous Fluid Shear Stress on the Protein Expression of OPG/RANKL in Osteoblast

作　　者：陈少龙[1] 赵良功[1] 滕元君[1] 陈孝琼[1] 姜金[1] 夏亚一[1] 汪静[1] 王翠芳[1] 安丽萍[1] 马靖琳[1] 

机构地区：[1]兰州大学第二医院骨科甘肃省骨关节疾病研究重点实验室,甘肃兰州730000

出　　处：《中国医学物理学杂志》2015年第1期120-123,133,共5页Chinese Journal of Medical Physics

基　　金：国家自然科学基金(81071478);甘肃省自然科学基金(1107RJZA144)

摘　　要：目的:观察两种不同加载模式的流体剪切力(周期性或持续性)对MC3T3-E1成骨细胞增殖以及骨保护素(OPG)、细胞核因子k B受体活化因子配体(RANKL)蛋白表达的影响,同时探讨影响成骨细胞功能的最佳流体剪切力加载模式。方法:对MC3T3-E1成骨细胞分别加载生理强度为12 dyn/cm2的周期性流体剪切力或持续性流体剪切力。其中,持续性模式采用12 dyn/cm2的流体剪切力,加载成骨细胞2 h,静息2 h;周期性流体剪切力采用12 dyn/cm2流体剪切力加载成骨细胞30 min,然后静息3 0min,如此往复循环4次,总的加力时间为2 h。最后,MTT法检测不同加载模式下成骨细胞的增殖情况,Westernblot检测两组成骨细胞OPG、RANKL蛋白的表达影响。结果:MTT结果显示,两种加载模式的流体剪切力均能够显著促进成骨细胞的增殖(P<0.05),与持续性流体剪切力相比,周期性流体剪切力更能有效的促进成骨细胞的增殖(P<0.05)。另外,两种模式的流体剪切力均能有效地调控MC3T3-E1成骨细胞OPG、RANKL的表达(P<0.05)。但与持续性流体剪切力相比,周期性流体剪切力对成骨细胞OPG蛋白的促进作用更为显著(41.34%±5.37%vs 80.42%±4.19%,P<0.05);对成骨细胞RANKL蛋白的抑制作用更为明显[(24.17±5.92)%vs(8.45±2.18)%,P<0.05]。结论:与持续流体剪切力相比,周期性流体剪切力对成骨细胞增殖以及OPG、RANKL蛋白有更显著的调节作用。Objective This study aims to investigate the effect of two different loading modes （cyclic or continuous） of fluid shear stress （FSS） on the proliferation of MC3T3-EI osteoblast and OPG/R protein expression. Also, we＇ll explore the superior loading mode of FSS affecting osteoblastic function. Methods Two different loading modes （cyclic and continuous） of FSS with the strength of 12 dyn/cm2 were loaded on MC3T3-E1 osteoblast. Among them, 12 dyn/cm2 continuous FSS was used for loading osteoblast for 2 hours and rested for 2 hours. 12 dyn/cm2 Cyclic FSS was loaded on osteoblast for 30 minutes and rested for 30 minutes, cycling four times （2 hours total）. Finally, MTT and Westernblot were used to detect the effect of different loading modes of FSS on osteoblast proliferation and the protein expression of OPG/RANKL,respectively. Result The result of MTT showed that two modes of FSS both significantly promoted the osteoblast proliferation （P 〈 0.05）, and cyclic FSS had better effect in osteoblast proliferation than continuous FSS（P 〈 0.05）. In addition, both modes of FSS could effectively regulate the expression of OPG and RANKL in MC3T3-EI osteoblast （P 〈 0.05）. However, compared with the continuous FSS, cyclic FSS had more significant effect of promotion on osteoblast OPG protein （41.34%±5.37% vs 80.42%±4.19%, P 〈 0.05）, but had more significant effect of inhabition on RANKL protein （24.17%±5.92% vs 8.45%±2.18%, P〈0.05）. Conclusion Compared with the continuous FSS, cyclic FSS has more significant effect of regulation on proliferation and the expression of OPG and RANKL in MC3T3-E1 osteoblast.

关 键 词：成骨细胞 流体剪切力 骨保护素 细胞核因子kB受体活化因子配体 

分 类 号：R687.1[医药卫生—骨科学]