Toll样受体4及环氧化酶2在大鼠动脉血栓中的表达及相关性

Expressions and relationship of Toll like receptor 4 and cyclooxygenase 2 in arterial thrombus in rats

目的:研究Toll样受体4(TLR-4)及环氧化酶2(COX-2)在大鼠动脉血栓中的表达及相互关系。方法:采用免疫组织化学(组化)技术检测大鼠动脉血栓的TLR-4、COX-2蛋白表达和分布情况。采用蛋白质印迹(Western blotting)方法,对建立颈动脉血栓形成模型后第1、3、7、14天(D1、D3、D7、D14实验组)及相应天数假手术组(D1、D3、D7、D14对照组)的Sprague Dawley(SD)大鼠(均n=10)外周血单个核细胞中COX-2及TLR-4表达水平进行检测,并分析上述因子水平之间的相关性。结果:免疫组化见实验组血栓中的炎性细胞TLR-4、COX-2染色呈不同程度阳性,其中D1、D3组阳性染色不明显,D7、D14组的切片阳性染色逐渐加强。蛋白质印迹结果显示实验组中TLR-4、COX-2蛋白表达量随血栓形成后天数的推移而逐渐升高,并在第14天达到了最高值,相关性分析结果表明,TLR-4与COX-2蛋白水平变化有明显相关性(r=0.831,P=0.007);而对照组TLR-4及COX-2表达量低。结论:COX-2、TLR-4蛋白表达水平在大鼠颈动脉血栓中明显增加,提示其与动脉血栓形成有关,可能参与了动脉血栓形成的过程,并且COX-2与TLR-4在蛋白水平的表达上具有一定相关性。

Objective To investigate the expressions of cyclooxygenase 2 （COX-2） and Toll like receptor 4（TLR-4） in arterial thrombus and analyze their relationship. Methods Immunohistochemistry was used to detect the expression and distribution of TLR-4 and COX-2 protein in arterial thrombus in rats. Western blotting was used to detect the expression levels of COX-2 and TLR-4 in peripheral blood monocytes of Sprague Dawley （SD） rats with carotid arterial thrombosis established （model group） and corresponding sham operation control group on D （day）l, D3, D7 and D14 after the establishment of arterial thrombosis, and the correlation between levels of COX-2 and TLR-4 was analyzed. Results Immunohistochemistry demonstrated that varying degrees of TLR-4, COX-2 staining in inflammatory cells of arterial thrombus were observed; the positive staining was weak in D1, D3 groups and gradually increased in D7, D14 group. Expression levels of TLR-4 and COX-2 in peripheral mononuclear cells were increased gradually with time and reached the highest value in D14 group; TLR-4 had a good correlation with COX-2 （r=0.831, P=0.007）. The expression levels of TLR-4 and COX-2 in peripheral mononuclear cells were low in sham operation control group. Conclusions Expressions of COX-2 and TLR-4 in carotid artery thrombus were increased significantly, suggesting it may be related with and involved in the process of arterial thrombosis. Correlation was found between expressions of COX-2 and TLR-4.