目的研究组织金属蛋白酶抑制因子3(TIMP-3)重组蛋白对MC3T3-E1成骨细胞凋亡的影响。方法细胞存活率和凋亡分别用MTT及ELISA检测。Fas,FasL,Bcl-2,Bax,caspase-3,caspase-8,细胞色素c的表达以及JNK,p38,细胞外信号调节激酶(ERK)1/2的磷酸化水平用Western印迹检测。结果MTT及ELISA检测提示TIMP-3降低MC3T3-E1细胞存活率,诱导MC3T3-E1细胞凋亡。TIMP-1增加Fas、FasL蛋白表达,对Bax、Bcl-2蛋白表达无影响,但可诱导细胞色素c的释放及caspase-8、caspase-3的活化。TIMP-3促进p38和ERK磷酸化,而PD098059(ERK阻断剂)和SB203580(p38阻断剂)消除p38和ERK的促凋亡作用。结论TIMP-3诱导MC3T3-E1细胞凋亡的信号途径为凋亡受体Fas介导,并有p38、ERK信号转导途径参与。 Objective To study the effect of tissue inhibitor of metalloproteinase 3 (TIMP-3) on the apoptosis of MC3T3-E1 osteoblasts. Methods Cell viability and apoptosis were detected by MTT and ELISA, respectively. The phosphorylation of Fas, FasL, Bcl-2, Bax, caspase-3, caspase-8 and cytochrome c and phosphorylation of JNK, p38 and extracellular signal-regulated kinase (ERK) 1/2 were detected by Western blotting. Results The results of MTT and ELISA showed that TIMP-3 decreased the survival rate of MC3T3-E1 cells and induced the apoptosis of MC3T3-E1 cells. TIMP-1 increased Fas and FasL protein expression, but had no effect on Bax and Bcl-2 protein expression, but could induce the release of cytochrome c and the activation of caspase-8 and caspase-3. TIMP-3 promotes p38 and ERK phosphorylation, whereas PD098059 (ERK blocker) and SB203580 (p38 blocker) abrogate the pro-apoptotic effects of p38 and ERK. Conclusion The signal pathway of TIMP-3-induced apoptosis in MC3T3-E1 cells is mediated by Fas receptor, and p38 and ERK signal transduction pathways are involved.