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作 者:陈少龙[1] 滕元君[1] 赵良功[1] 姜金[1] 崔兆辉[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,兰州730000
出 处:《世界科技研究与发展》2014年第4期415-419,共5页World Sci-Tech R&D
基 金:国家自然科学基金(81071478);甘肃省自然科学基金(1107RJZA144)资助
摘 要: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.
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