ERK5阻断剂BIX02188介导的流体剪切力对成骨细胞OPG、RANKL mRNA表达的影响

Fluid Shear Stress Mediates OPG /RANKL mRNA Expression in MC3T3-E1 Cell via ERK5 Inhibitor BIX02188

目的观察在流体剪切力(Fluid Shear Stress,FSS)作用下,成骨细胞(MC3T3-E1)中骨保护素(osteoprotegerin,OPG)和细胞核因子κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)mRNA的表达情况,以及探讨ERK5信号通路在其中的作用。方法用不同浓度的ERK5特异性阻断剂BIX02188干预MC3T3-E1成骨细胞,用MTT比色法检测490nm吸光度值,观察成骨细胞的增殖情况,并检测ERK5 mRNA的表达情况。给成骨细胞加载生理强度为12 dyne/cm^2的流体剪切力,观察OPG、RANKL mRNA的表达。结果浓度为15μM的ERK5特异阻断剂BIX02188能够有效的抑制ERK5 mRNA的表达;生理强度为12 dyne/cm^2的FSS能够显著地促进OPG mRNA表达(P<0.05),降低RANKL mRNA表达(P<0.05);当BIX02188干预成骨细胞后,FSS对成骨细胞OPG、RANKL mRNA的影响明显减弱(P<0.05)。结论 ERK5特异阻断剂BIX02188能够有效介导FSS对成骨细胞OPG、RANKL mRNA的表达。

Objective To observe OPG/RANKL mRNA expression in MC3T3-E1 cell induced by fluid shear stress,as well as to study the effect of ERK5 signaling pathways in it.Methods A specific inhibitor of ERK5 （BIX02188）was used to inhibit the expression of ERK5 mRNA.MTT assay and real-time PCR were used to investigate the cellular proliferation and ERK5 /OPG/RANKL mRNA expression respectively.Results The expression of ERK5 mRNA was significantly inhibited by BIX02188 at the concentration of 15 μmmol/L （P〈0.05）.After cultivating the MC3T3-E1 with BIX02188,osteoblastic proliferation was significantly inhibited （P 〈0.05）.Moreover,FSS significantly increased the expression of OPG mRNA （P〈0.05）and reduced the expression of RANKL mRNA（P 〈0.05）.However,ERK5 down-regulated OPG/RANKL mRNA expression induced by FSS （P〈0.05）.Conclusion ERK5 inhibitor （BIX02188）can significantly mediate the expression of OPG/RANKL mRNA induced by fluid shear stress.