辛伐他汀防治人工关节无菌性松动的实验研究  

EXPERIMENTAL STUDY ON SIMVASTATIN IN PREVENTION AND TREATMENT OF ASEPTIC LOOSENING OF PROSTHESIS

作　　者：王真[1,2] 高希武[1,3] 孙克宁[1] 金群华[3] 

机构地区：[1]宁夏医科大学研究生院,银川750004 [2]宁夏回族自治区人民医院骨科 [3]宁夏医科大学附属医院骨3科

出　　处：《中国修复重建外科杂志》2010年第5期544-547,共4页Chinese Journal of Reparative and Reconstructive Surgery

摘　　要：目的通过观察辛伐他汀对钛颗粒刺激人外周血单个核细胞(peripheral blood mononuclear cell,PBMC)而释放TNF-α及单核细胞趋化蛋白1(monocyte chemoattractant protein1,MCP-1)的影响,分析该过程中p42/p44激酶磷酸化的情况,评价辛伐他汀在防治人工关节无菌性松动中的作用。方法取成年健康志愿者外周血45mL,分层梯度离心法分离获得PBMC,制成密度为5×108个/mL的细胞悬液,根据培养液不同分成5组。A组:加入3mL RPMI-1640培养液及1mL0.01%钛颗粒悬液;B、C、D、E组:同A组试剂外,分别加入2mL浓度为1×10-5、1×10-6、1×10-7mmol/L辛伐他汀和20μmol/L U0126。取培养24h后上清液检测TNF-α及MCP-1含量。取培养2h后PBMC,按A～E组分组方法分别用不同培养液预处理60min,钛颗粒分别刺激培养30、60min,采用Western blot检测细胞外信号调节激酶(extracellular signal-regulated kinases,ERK1/2)的表达水平。结果A、B、C、D、E组TNF-α含量分别为1.1155±0.2436、0.6936±0.3543、0.6957±0.3873、0.7164±0.4789、0.2635±0.1016;MCP-1分别为1.4210±0.1053、0.9151±0.4113、1.0035±0.4642、1.1020±0.3539、0.2713±0.1451。A组TNF-α及MCP-1含量明显高于其他组,差异有统计学意义(P<0.05);E组与B、C、D组比较差异有统计学意义(P<0.05),B组与C、D组比较差异有统计学意义(P<0.05),C、D组间比较差异无统计学意义(P>0.05)。Western blot检测显示钛颗粒刺激培养30、60min各组均见ERK1/2表达;刺激培养30min时A、B、C、D、E组ERK1/2含量较低;60min时A、B、C、D、E组ERK1/2含量分别为1.6121±0.0682、1.0781±0.0728、1.2687±0.2231、1.4397±0.1801、0.7320±0.1104,组间比较差异均有统计学意义(P<0.05)。结论ERK1/2通路是磨屑诱导巨噬细胞生成TNF-α和MCP-1的主要通路,辛伐他汀可抑制此通路,进而有效防止关节磨屑诱导的人工关节松动。Objective To evaluate the mechanisms of p42/p44 kinase phosphorylation in cell models and to investigate the effect of simvastatin on the prevention and treatment of aseptic loosening of prosthesis by observing the in？ uence of simvastatin on the levels of tumor necrosis factor α （TNF-α） and monocyte chemoattractant protein 1 （MCP-1） of human peripheral blood mononuclear cell （PBMC） challenged with titanium particles. Methods PBMC from 45 mL peripheral blood of healthy adult voluntary donators,were separated and cultured,and divided into 5 groups according to different culture medium：group A,PBMC and titanium particles; group B,PBMC and titanium particles with 1 × 10^-5 mol/L simvastatin; group C,PBMC and titanium particles with 1 × 10^-6 mol/L simvastatin; group D,PBMC and titanium particles with 1 × 10^-7 mol/L simvastatin; and group E,PBMC and titanium particles with the extracellular signal-regulated kinase （ERK1/2） inhibitor U0126. The contents of TNF-α and MCP-1 were tested by ELISA after 24 hours of culture. PBMC were pretreated with different medium grouping as groups A,B,C,D,and E for 60 minutes,and were challenged with titanium particles for 30 minutes and 60 minutes,then the level of ERK1/2 expression was tested by Western blot. Results In groups A,B,C,D,and E,the absorbance （A） values of TNF-α were 1.115 5 ± 0.243 6,0.693 6 ± 0.354 3,0.695 7 ± 0.387 3,0.716 4 ± 0.478 9,and 0.263 5 ± 0.101 6,respectively; and the A values of MCP-1 were 1.421 0 ± 0.105 3,0.915 1 ± 0.411 3,1.003 5 ± 0.464 2,1.102 0 ± 0.353 9,and 0.271 3 ± 0.145 1,respectively. The levels of TNF-α and MCP-1 in group A were significantly higher than others,showing significant differences （P〈 0.05）. There were signif icant differences between group E and groups B,C,and D （P〈 0.05）,between group B and groups C,D （P 〈0.05）; no significant difference between group C and group D （P〉0.05）. Western blot results showed the expression of ERK1/2 in all groups at 30 minutes and 60 minu

关 键 词：辛伐他汀 人工关节无菌性松动 骨溶解 细胞外信号调节激酶 

分 类 号：R687.4[医药卫生—骨科学]