流体剪切力对MC3T3-E1细胞OPG、RANKL蛋白表达的实验研究

Effect of Fluid Shear Stress on the Protein Expression of OPG and RANKL in MC3T3-E1 Cells

目的探讨流体剪切力(fluid shear stress,FSS)作用下,两种调节骨骼重建的重要分子骨保护素(osteoprotegerin,OPG)和细胞核因子κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)的蛋白表达情况。方法采用体外模型对MC3T3-E1细胞加载流体剪切力,细胞经不同时间加力后(0,30,60,90,120min),对细胞分别进行染色和裂解,运用免疫荧光和蛋白印迹法对OPG和RANKL的蛋白表达水平进行定量分析。结果 FSS作用30,60,90,120min后能够显著增加OPG的蛋白表达(P<0.05),减少RANKL的蛋白表达(P<0.05)。两者共同作用使得OPG/RANKL值显著增高(P<0.05)。结论流体剪切力刺激提示OPG/RANKL的比值可能在成骨细胞和破骨细胞联合调节骨骼形成和吸收的过程中起着重要的调节作用。

Objective Explore the effect of fluid shear stress（FSS）on the protein expressions of two important regulators of bone remodeling,osteoprotegerin（OPG）and receptor activator of NF-κB ligand（RANKL）.Methods We developed an in vitro model to load FSS on MC3T3-E1 cells in various durations（0,30,60,90,and 120 min）,the levels of OPG and RANKL were quantified using immunofluorescence and Western Blotting when cells were lysed.Results Protein expression of OPG was significantly increased and RANKL was significantly reduced at 30,60,90,and 120 min groups（P〈0.05）,which resulted in a significant raise of OPG/RANKL ratio（P〈0.05）.Conclusions Ratio of OPG to RANKL can be an important regulator in the bone formation and resorption regulated by osteoblastic and osteoclastic mechanisms.