钛颗粒刺激巨噬细胞而释放肿瘤坏死因子对人工关节松动作用.docVIP

钛颗粒刺激巨噬细胞而释放肿瘤坏死因子对人工关节松动作用【摘要】 目的 分析钛颗粒刺激巨噬细胞而释放的肿瘤坏死因子在人工髋关节术后引起骨溶解而假体松动的分子生物学机制。方法 采用钛颗粒刺激巨噬细胞而释放TNF-α模型及酶联免疫吸附剂测定、蛋白质印迹、凝胶迁移率实验方法来研究其生物分子学机制。结果 鼠类巨噬细胞受钛颗粒刺激而释放TNF-α，而JNK抑制剂 SP600125和 NF-κB抑制剂MG132 明显抑制了钛颗粒诱导的TNF-α释放，但p38抑制剂无此作用，甚至，钛颗粒诱导巨噬细胞而激活AP-1 和 NF-κB。结论 钛颗粒诱导的TNF-α释放与JNK/AP-1及NF-κB通路有关。? 【关键词】人工髋关节置换术；钛颗粒；骨溶解; 肿瘤坏死因子 ?? The effect of TNF-α release in macrophages stimulated with titanium particles on the aseptic loosening of artificial joints JIN Shan.Yanbian the Second Peoples Hospital,Jilin133001，China ? 【Abstract】 Objective To analyse the mechanism ofosteolysis after total hip replacement by tumor necrosis factor-α(TNF-α)release in macrophages stimulated with particles.Methods A model of TNF-α release by stimulating macrophages with titanium particles was made and study the mechanism of TNF-α release by ELISA/West blot/EMSA.Results TNF-α was released in murine macrophage stimulaged by titanium(Ti)particles and specific inhibitors of JNK(SP600125)and NF-κB(MG132),but not p38(SB203580)dramatically inhibited Ti-stimulated TNF-α release.Moreover,transcriptional activation of AP-1 and NF-κB was also induced by Ti particles.Conclusion These results demonstrate that Ti particle-induced TNF-α release is mediated through JNK/AP-1 as well as NF-κB pathways.? 【Key words】THR; Ti particles; osteolysis; TNF-α ? 人工髋关节置换术后假体周围的骨溶解而引起的假体松动是人工关节术后失败的常见原因。由于假体长时间磨损及腐蚀生成的颗粒碎片，导致假体周围形成肉芽肿假膜，这些假膜组织包含的巨噬细胞、纤维母细胞、巨细胞及破骨细胞等[1-2]受颗粒碎片刺激而产生有关骨溶解及假体松动的炎症因子如TNF-α、IL-1β、IL-6、IL-8、IL-11及转化生长因子-β[3-4]的相互作用，使破骨细胞被激活而吸收骨组织，导致骨溶解及假体松动。? 在这些炎症因子中，TNF-α是很重要的骨溶解因子。Kimble 等已证明TNF-α可以增强成骨细胞NF-κB配位子(RANKLE)受体及巨噬细胞-集落刺激因子(M-CSF)受体活性[5]。而且，Ingham 等发现TNF-α又直接促进前破骨细胞分化并激活成熟的破骨细胞吸收骨组织[6]。与此同时,Merkel 等通过去除肿瘤坏死因子受体(TNFR)基因的动物颅骨与PMMA 颗粒接触，发现无骨溶解现象[7]。这些结果显示，TNF-α是人工关节术后磨损颗粒导致的骨溶解重要细胞因子。但至今,尚未报告与此相关的分子生物学机制。本次实验,通过鼠巨噬细胞受钛颗粒刺激而释放的TNF-α模型来分析TNF-α合成的分子生物学机制。? 1 材料与方法? 1．1 材料? 1．1．1 DMEM培养基，牛胎血清(FBS)来自于Gibco Invitrogen 公司(Rockville,MD)。PD98059、SP600125、MG132 来自于Calbioch