人工关节假体磨损颗粒对巨噬细胞游走抑制因子表达的影响  

Effect of wear particles from joint prosthesis on the expression of macrophage migration inhibiting factor

作　　者：潘孝云[1] 许心弦[1] 张先龙[2] 毛昕 温宏[1] 张宇[1] 

机构地区：[1]温州医学院附属第二医院,浙江温州325027 [2]上海市第六人民医院,上海200211

出　　处：《中医正骨》2013年第1期14-18,共5页The Journal of Traditional Chinese Orthopedics and Traumatology

基　　金：浙江省温州市科技计划项目(Y20120027)

摘　　要：目的:观察人工关节假体磨损颗粒对巨噬细胞游走抑制因子表达的影响。方法:配制钛颗粒悬液,并培养小鼠巨噬细胞。将培养的第3代小鼠巨噬细胞分别进行以下实验:①将细胞分为4组,A组不作任何处理,B组加入浓度为0.01%的钛颗粒;C组加入浓度为0.05%的钛颗粒,D组加入浓度为0.1%的钛颗粒。培养24 h后分别用酶联免疫吸附剂测定法和聚合酶链式反应法检测各组的巨噬细胞游走抑制因子蛋白和巨噬细胞游走抑制因子mRNA的含量。②将细胞分2组,对照组不作任何处理,观察组加入浓度为0.1%的钛颗粒。分别在实验开始后0 h、6 h、12 h、24 h和36 h对2组的不同培养孔的细胞用酶联免疫吸附剂测定法和聚合酶链式反应法检测各组的巨噬细胞游走抑制因子蛋白和巨噬细胞游走抑制因子mRNA的含量。③将细胞分为4组,Ⅰ组不作任何处理,Ⅱ组加入浓度为0.1%的钛颗粒,Ⅲ组加入浓度为100μmol.mL-1的吡咯烷二硫代氨基甲酸盐,Ⅳ组先加入浓度为100μmol.mL-1的吡咯烷二硫代氨基甲酸盐,1 h后再加入浓度为0.1%的钛颗粒。培养24 h后用酶联免疫吸附剂测定法检测各组的巨噬细胞游走抑制因子蛋白含量。④将细胞分为2组,a组不作任何处理,b组加入浓度为0.1%的钛颗粒。分别在实验开始后0 h、0.5 h、1 h、3 h和6 h对2组的不同培养孔的细胞用酶联免疫吸附剂测定法测定磷酸化p65的含量。结果:①钛颗粒浓度对巨噬细胞游走抑制因子表达的影响:各组小鼠巨噬细胞巨噬细胞游走抑制因子蛋白和巨噬细胞游走抑制因子mRNA含量的组间比较,差异均有统计学意义(F=207.158,P=0.000;F=64.955,P=0.000)。A组巨噬细胞游走抑制因子蛋白和巨噬细胞游走抑制因子mRNA的含量[(3.93±0.11)ng.mL-1,(0.03±0.01)]与B组[(4.21±0.27)ng.mL-1,(0.10±0.01)]比较,差异无统计学意义(P=0.167;P=0.223);A组小于C组[(6.56±0.27)ng.mL-1,(0.25±0.09)],差异有统计学意义(Objective：To observe the effects of wear particles from joint prosthesis on the expression of macrophage migration inhibiting factors（MMIF）.Methods：Suspension with titanium particles was prepared and mice macrophages were cultured.The following experimentations were respectively carried on the third generation of mice macrophages.①The cells were divided into 4 groups,cells in group A were added with nothing,while cells in group B,C and D were added with titanium particles suspension（0.01%,0.05% and 0.1%,respectively）.The contents of MMIF protein and MMIF mRNA of the 4 groups were respectively detected through enzyme linked immunosorbent assay（ELISA）and polymerase chain reaction（PCR）after culturing for 24 hrs.②The cells were divided into 2 groups,cells in control group were added with nothing,while cells in observation group were added with 0.1% titanium particles suspension.The contents of MMIF protein and MMIF mRNA of the 2 groups were respectively detected through ELISA and PCR at 0,6,12,24 and 36 hrs from the beginning of the experimentation.③The cells were divided into 4 groups,cells in groupⅠwere added with nothing,and cells in groupⅡwere added with 0.1% titanium particles suspension and groupⅢwere added with pyrrolidine dithiocarbamate（PDTC）（100 μmol/ml）,while cells in groupⅣwere firstly added with PDTC（100 μmol/ml）and titanium particles suspension（0.1%）one hr later.The contents of MMIF protein of the 4 groups were detected through ELISA after culturing for 24 hrs.④The cells were divided into 2 groups,cells in group a were added with nothing,while cells in group b were added with 0.1% titanium particles suspension.The contents of phosphorylation p65 of the 2 groups were respectively detected through ELISA at 0,0.5,1,3 and 6 hrs from the beginning of the experimentation.Results：①Effect of titanium particles suspension concentration on MMIF expression：There were statistical differences in the contents of MMIF protein and MMIF mRNA of macrophages amon

关 键 词：人工关节 巨噬细胞游走抑制因子 NF-ΚB 动物实验 

分 类 号：R318.1[医药卫生—生物医学工程]