金属离子刺激人单核细胞分泌TNF-α并诱导其凋亡与Caspase-3激活的实验研究  

RELEASE OF TNF-α AND ACTIVATION OF CASPASE-3 DURING APOPTOSIS OF HUMAN MONONUCLEAR CELL INDUCED BY METAL ION

作　　者：熊建卫[1] 戴闽[1] 郝亮[1] 漆启华[1] 詹平[1] 

机构地区：[1]南昌大学第一附属医院骨二科,南昌330006

出　　处：《中国修复重建外科杂志》2009年第3期299-302,共4页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：南昌大学学科交叉基金资助项目(Z03722)~~

摘　　要：目的观察人工关节磨损产物Co2+、Cr3+刺激下人单核细胞生物反应过程中TNF-α的表达,以及人单核细胞凋亡与Caspase-3的变化,探讨Co2+、Cr3+引发假体周围骨溶解的作用机制。方法将CoCl2粉末与CrCl3粉末溶于无菌注射用水,分别配制成浓度为10mg/L和500mg/L的CoCl2溶液和CrCl3溶液。从健康志愿者外周血分离培养单核细胞,调整浓度为4×106个/培养皿,根据培养液不同,分为3组。A组生理盐水作为对照,B组CoCl2溶液,C组CrCl3溶液。于培养12、24及48h后吸取细胞上清液,ELISA法检测TNF-α含量,TUNEL检测细胞凋亡情况,比色法测定Caspase-3活性。结果各时间点B、C组TNF-α含量及Caspase-3活性均高于A组(P<0.05),分别于24h及48h达峰值。各时间点3组均存在TUNEL染色阳性细胞,细胞核固缩深染,细胞体积皱缩,胞膜完整。各时间点B、C组细胞凋亡率与A组比较,差异有统计学意义(P<0.05)。Caspase-3活性与细胞凋亡率成正相关(r=0.765)。结论Co2+、Cr3+均能刺激人单核细胞释放TNF-α并诱导其凋亡;细胞凋亡参与了假体周围骨溶解的病理过程,且可能与Casepase-3的激活有关。Objective To observe the human mononuclear cell releasing TNF-α and the activation of Caspase-3 during apoptosis after stimulated by Co2^＋ and Cr3^＋,to discuss the mechanism of arti cial joint wear production metal ion on aseptic loosening. Methods CoCl2 powder and CrCl3 powder were dissolved by asepsis inject water,preparing solution for 10 mg/L and 500 mg/L,respectively. Mononuclear cells were acquired from peripheral blood,4×106/culture dish. According to the di erence of culture solution,the cells were divided into 3 groups. Group A： mononuclear cell was culture with normal saline as control; group B： mononuclear cell was cultured with CoCl2 solution; group C： mononuclear cell was cultured with CrCl3 solution. The production of TNF-α was assessed by ELISA,the activation of Caspase-3 was measured by colorimetric assay and the apoptotic cell was detected by TUNEL assays at 12,24 and 48 hours after co-cultured respectively. Results The concentration of TNF-α and theffexpression of Caspase-3 in groups B and C were higher than those in group A （P 〈 0.05）,and reached the peak level at 24,48 hours,respetively. The TUNEL positive cells were detected in the all groups,nucleus was pyknotic and darker-staining,cell body was crinkle and cell membrane was integrity. There were signi cant di erences in the apoptosis rate between groups B,C and group A （P 〈 0.05）. And the activation of Caspase-3 increased and had the positive correlation with the apoptosis rate （r=0.765）. Conclusion Co2^＋ and Cr3^＋ ions can stimulate human mononuclear cell to release TNF-α and induce human mononuclear cell apoptosis,which result in periprosethetic osteolysis and related to activation of Caspase-3.

关 键 词：金属离子 人工关节无菌性松动 TNF-Α CASPASE-3 细胞凋亡 

分 类 号：R687.4[医药卫生—骨科学]