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Differential expression of cyclooxygenase-1 and cyclooxygenase-2 in the cornea during wound healing

机译:伤口愈合过程中角膜上环氧合酶-1和环氧合酶-2的差异表达

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Cyclooxygenases (COXs) are the key enzymes in the production of prostaglandins (PGs) and exist in two isoforms. Isoform 1 (COX-1) is constitutively expressed in most tissues, whereas cyclooxygenase-2 (COX-2) is rapidly induced by a variety of different stimuli. In this study, we have quantitatively analyzed mRNA expression of COX-1 and COX-2 and protein distribution during corneal reparative processes after wound. Total RNA was isolated from cornea samples of New Zealand rabbits that had been subjected to corneal wound by mechanical brush scraping. Quantification of RT-PCR results was made by using a DNA mimic approach. The localization and expression of the enzymes was studied by immunocytochemistry and Western blotting. In normal corneas COX-1 is expressed throughout the cornea in the whole tissue, while COX-2 is strongly expressed in stromal keratocytes. Following injury, COX-2 levels drastically increase and, at least in the epithelium, COX-2 becomes the predominant isoform of cyclooxygenases at an early stage of healing. Moreover, in the epithelium COX-2 is expressed predominantly by those cells close to the wound. These cells become migratory and move toward the injured area. In contrast, COX-1 levels remain unaffected in all corneal tissues. The system returns to the pre-injury state in about 24 h. Thus, the expression of COX-2 in the corneal epithelium during wound repair is tightly regulated both temporally and spatially. (C) 2003 Elsevier Ltd. All rights reserved
机译:环氧合酶(COXs)是前列腺素(PGs)生产中的关键酶,并以两种同工型存在。异构体1(COX-1)在大多数组织中组成性表达,而环氧合酶2(COX-2)可通过多种不同的刺激快速诱导。在这项研究中,我们定量分析了伤口后角膜修复过程中COX-1和COX-2的mRNA表达以及蛋白质分布。从已经通过机械刷刮擦经受角膜伤口的新西兰兔的角膜样品中分离总RNA。 RT-PCR结果的定量是通过使用DNA模拟方法进行的。通过免疫细胞化学和蛋白质印迹研究了酶的定位和表达。在正常角膜中,COX-1在整个组织的整个角膜中表达,而COX-2在基质角膜细胞中强烈表达。损伤后,COX-2水平急剧增加,并且至少在上皮中,COX-2在愈合的早期成为环氧合酶的主要同工型。此外,在上皮中,COX-2主要由靠近伤口的那些细胞表达。这些细胞迁移并移向受伤区域。相反,COX-1水平在所有角膜组织中均不受影响。系统在约24小时内恢复到受伤前的状态。因此,在伤口修复过程中,角膜上皮中COX-2的表达在时间和空间上均受到严格调节。 (C)2003 Elsevier Ltd.保留所有权利

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