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目的探讨缺血和缺血-再灌注状态下对大鼠心肌基质金属蛋白酶-1(MMP-1)的影响。方法取正常大鼠心脏和通过钳夹大鼠心脏冠状动脉旋支制造缺血-再灌注模型的心肌组织,利用免疫组织化学、Westernblotting技术和计算机图像处理系统,对正常和缺血-再灌注大鼠心脏MMP-1的表达进行形态学观察,并对其含量进行定量分析。结果1.免疫组织化学反应显示正常心脏的MMP-1主要分布于心肌的细胞间质。纤维细胞、血管平滑肌细胞、毛细血管内皮细胞均呈强阳性反应。缺血30min心肌MMP-1反应无明显变化,缺血60min,酶反应浓度明显增加,缺血-再灌注30min,MMP-1的阳性反应增强,缺血-再灌注60min后,MMP-1反应区出现大面积融合。2.定量分析显示,缺血30min,MMP-1无显著性改变(P>0.05);缺血60min,出现显著性改变(P<0.05);缺血-再灌注30min,有显著性改变(P<0.01);缺血-再灌注60min,出现高度显著性改变(P<0.001)。3.Western blotting显示,缺血30min、60min肉眼观察无明显变化。缺血-再灌注30min,MMP-1条带开始增宽,缺血-再灌注60min,MMP-1条带显著增宽。结论1.正常情况下MMP-1由毛细血管内皮、心内膜的内皮、小动脉平滑肌细胞分泌;在缺血、再灌注情况下心肌细胞具有分泌MMP-1的潜能。2.随心肌缺血时间的延长,MMP-1的含量逐渐增多,再灌注可进一步增加MMP-1的含量,这可能是心肌缺血-再灌注后胶原迅速破坏的主要原因之一。
Objective To investigate the effect of ischemia / reperfusion on the expression of matrix metalloproteinase-1 (MMP-1) in rat myocardium. Methods The myocardial tissue of ischemia-reperfusion model was obtained from the normal rat heart and the coronary artery branches of the rat heart by clamping. Using immunohistochemistry, Westernblotting and computer image processing system, the normal and ischemia-reperfusion Morphological observation was performed on the expression of MMP-1 in rat heart, and its content was quantitatively analyzed. Results 1. Immunohistochemistry showed that the normal heart MMP-1 is mainly distributed in the myocardial interstitial. Fibroblasts, vascular smooth muscle cells, capillary endothelial cells showed a strong positive reaction. After ischemia-reperfusion for 30min, there was no significant change of MMP-1 in the myocardium. The activity of MMP-1 increased 60min after ischemia, and the positive reaction of MMP-1 increased after ischemia-reperfusion 30min. A large area of convergence. Quantitative analysis showed that there was no significant change of MMP-1 at 30min after ischemia (P> 0.05), significant change at 60min after ischemia (P <0.05), and at 30min after ischemia-reperfusion <0.01). There was a highly significant change at 60 min after ischemia-reperfusion (P <0.001). Western blotting showed that there was no significant change in macroscopic observation at 30min and 60min after ischemia. After ischemia-reperfusion for 30min, MMP-1 began to broaden, and 60min after ischemia-reperfusion, MMP-1 band widened obviously. MMP-1 is normally secreted by the endothelium and endothelium of the endothelium and arterioles in the smooth muscle cells of the arterioles. In the condition of ischemia and reperfusion, MMP-1 has the potential to secrete MMP-1. With the prolongation of myocardial ischemia, the content of MMP-1 gradually increased and reperfusion could further increase the content of MMP-1, which may be one of the main reasons for the rapid destruction of collagen after myocardial ischemia-reperfusion.