大鼠心肌缺血再灌注损伤组织中MMP-2、9/TIMP-1、2mRNA表达比值的变化及意义

MMP-2,9/TIMP-1,2 mRNA expression ratio changes in myocardiam ischemia reperfusion tissues of rat,and their significance

目的探讨心肌缺血再灌注损伤(MIRI)组织中基质金属蛋白酶-2、9(MMP-2、9)和组织金属蛋白酶抑制因子-1、2(TIMP-1、2)基因mRNA表达的变化规律及其临床意义。方法建立大鼠MIRI模型,采用HE、RTPCR、免疫组化、酶谱分析等方法,检测MIRI不同时相中MMP-2、9和TIMP-1、2 mRNA表达变化。结果在MIRI过程中MMP-2、9的mRNA和蛋白相对表达量呈升高趋势,二者在MIRI 8 h时表达量达高峰,与对照组(sham组)相比具有非常显著性差异(P<0.01)。TIMP-1、2的mRNA相对表达量在整个再灌注过程中增高,与sham组相比有显著性差异(P<0.05),而蛋白水平的相对表达量与sham组相比无统计学差异(P>0.05)。MMP-2、9/TIMP-1、2 mRNA表达比值趋于失衡。酶活性MMP-2在MIRI 4 h达高峰(16832.67±727.40),MMP-9在MIRI 12 h达高峰(6534.85±421.50),与sham组和MIRI 30 min组相比,二者均有非常显著性差异(P<0.01)。结论 MMP-2、9/TIMP-1、2mRNA表达比值失衡是MIRI的重要指标之一。

Objective To study the change regularity and clinical significance of MMP-2,9/TIMP-1,2 mRNA expression in myocardium ischemia-reperfusion injury（MIRI）. Methods A rat MIRI model was established. The Changes of MMPs-2,9/TIMP-1,2 mRNA expression in rat myocardium during ischemia-reperfusion were detected by HE,RT-PCR,IHC,Zymogram analysis. Results The expression of MMP-2,9 mRNA and protein tended to increase during ischemia reperfusion. There was significant difference of MMP-2,9 mRNA expression in MIRI 8 h compared with sham group and at MIRI 30 min （P〈0.01）,respectively. The expression of TIMP-1,2 mRNA tended to increase during ischemia reperfusion. There was significant difference compared with sham group and at MIRi 30rain （P〈0.05）,but the expression of TIMP-1,2 protein tended to decrease during ischemia reperfusion. There was not any significant differences compared with sham group and at MIRI 30 min （P〉0.05）. MMPs/TIMPs ratio was loss of equilibrium. Zymogram analysis showed：MMP-2 reached peak at 4 h,MMP-9 reached peak at 12h,compared with sham and at MIRI 30 min （P〈0.01）. Conclusion MMP-2,9/TIMP-1,2 ratio disequilibrium can induce myocardium ischemia-reperfusion injury thus being a significant reason in during ischemia reperfusion.