ERK信号通路对钛合金诱导成骨细胞RANKL、OPG表达的作用  

The role of ERK signaling pathway in RANKL and OPG expression induced by titanium alloy particles in mouse osteoblasts

作　　者：谢守祥[1] 许铁[2] 刘林[2] 姚爱明[2] 刘凯[2] 刘筱[2] 

机构地区：[1]徐州医学院研究生学院2009级,江苏徐州221002 [2]徐州医学院附属医院急救中心,江苏徐州221002

出　　处：《徐州医学院学报》2012年第6期363-366,共4页Acta Academiae Medicinae Xuzhou

基　　金：国家自然科学基金重点项目（50535050）

摘　　要：目的探讨钛合金颗粒对体外培养的小鼠成骨细胞核因子-κB受体活化因子配体（RANKL）及骨保护素（OPG）基因表达和蛋白分泌的影响，以及细胞外信号调控激酶1／2（ERKl／2）信号通路在其中的作用。方法体外培养成骨细胞。实验分3组：对照组（A组）；钛合金组（B组）：给予浓度为0．1g／L钛合金颗粒干预；抑制剂组（c组）：给予钛合金颗粒＋ERK信号通路抑制剂P1）98059，每组6个样本。采用RT—PCR和ELISA法分别检测成骨细胞RANKL、OPG基因和蛋白表达。结果3组各个时间点均可见RANKL、OPG的表达；但B组RANKL、OPGmRNA的表达和蛋白分泌量以及RANKL／OPG比值显著高于A组（P〈0．01）；而c组RANKL、OPGmRNA的表达和蛋白分泌水平以及RANKL／OPG比值的增幅显著小于B组（P〈0．01）。结论钛合金颗粒可刺激成骨细胞RANKLmRNA、OPGmRNA的表达并促进RANKL、OPG蛋白的分泌，增加RANKL／OPG比值；ERK信号通路抑制剂PD98059可一定程度抑制RANKL和OPG的表达，降低RANKL／OPG比值，对减少假体周围骨溶解的发生可能起重要作用。Objective To investigate the effects of titanium alloy particles on gene expression and protein secretion of receptor activator of nuclear factor -κB ligand （RANKL） and osteoprotegerin （OPG） in mouse osteoblasts in vitro and the function of ERK signaling pathway in the above process. Methods Osteoblasts were cultured in vitro,and divided in- to three groups according to the culture solution used： group A is control group, osteoblasts were cultured with normal medium; group B, osteoblasts were cultured with 0.1 g/L titanium alloy particles medium; group C, osteoblasts were cul- tured with 0.1 g/L titanium alloy particles medium and inhibitor of ERK signaling inhibitor （PD98059）, there were six samples in each group. After 24 h and 48 h, RT - PCR and ELISA were performed to detect the expression of RANKL and OPG. Results The mRNA and protein expression of RANKL, OPG could be detected in three groups at 24 h and 48 h after culture. The mRNA expression and protein secretion of RANKL and OPG in group B was significantly higher than in group A （P 〈 0.01 ）, meanwhile, compared with group B, the mRNA expression and protein secretion of RANKL, OPG and the ratio of RANKL to OPG in group C was decreased significantly （ P 〈 0.01 ）. Conclusion Titanium alloy particles can up - regulate mRNA and protein expression of RANKL and OPG from osteoblasts. Inhibitor of ERK signaling （PD98059） can partly inhibite the expression of RANKL and of OPG, decrease the ratio of RANKL to OPG, which may play an important role in decreasing the occurrence of the periprosthetic osteolysis..

关 键 词：ERK1/2信号通路 成骨细胞 钛合金颗粒 细胞核因子-κB受体活化因子配体 骨保护素 

分 类 号：R641[医药卫生—外科学]