Expression of matrix metalloproteinases in wound healing after glaucoma filtration surgery in rabbits
Expression of matrix metalloproteinases in wound healing after glaucoma filtration surgery in rabbits
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DOI：
10.1159/000109987
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发表时间：
2007-01-01
影响因子：
2.1
通讯作者：
Sasaki, Hiroshi
中科院分区：
文献类型：
--
作者：
Shima, Ichirou;Katsuda, Shogo;Sasaki, Hiroshi
目的：目的探讨兔青光眼滤过术后创面基质金属蛋白酶（MMP）的蛋白和mRNA表达、明胶酶活性及MMP活性定位。方法：30只新西兰大白兔，分别于术后1、3、7、14、120天摘除60只眼，进行实验。分别采用免疫组织化学和激光捕获显微切割/实时荧光定量RT-PCR分析蛋白质和mRNA表达。采用明胶酶谱法分析其明胶酶活性，并采用原位酶谱法研究其定位。结果如下：免疫组化检测MMP-1、MMP-2、MMP-3、MMP-9和MT 1-MMP在创面中的表达，以术后3d最为明显。MMP阳性细胞主要为巨噬细胞。RT-PCR检测MMP-9和MT 1-MMP mRNA的表达。在手术后3天和7天的伤口中检测到对应于proMMP-2和MMP-2的活性形式的明胶溶解活性。原位酶谱定位在伤口部位的明胶分解活性。这些活动几乎完全取消了MMP抑制剂，表明明胶分解活性属于金属蛋白酶。结论：MMP，特别是MMP-2/MT 1-MMP，在青光眼滤过手术后伤口愈合过程中的细胞外基质降解中起重要作用，并且可能代表青光眼滤过手术后治疗干预的重要靶点。版权所有（C）2007 S. Karger AG，巴塞尔。
Purpose: To investigate the protein and mRNA expressions of matrix metalloproteinases (MMPs), gelatinolytic activity and localization of MMP activity in wounds after glaucoma filtration surgery in rabbits. Methods: Sixty eyes of 30 rabbits were removed 1, 3, 7, 14 and 120 days after the surgery and used for this experiment. Protein and mRNA expressions were analyzed by immunohistochemistry and laser capture microdissection/real-time RT-PCR, respectively. The gelatinolytic activity was analyzed by gelatin zymography and the localization was studied using in situ zymography. Results: By immunohistochemistry, expression of MMP-1, MMP-2, MMP-3, MMP-9 and MT1-MMP was detected in the wounds, most markedly 3 days after the surgery. MMP-positive cells were predominantly macrophages. Expression of MMP-9 and MT1-MMP mRNAs was verified by RT-PCR. Gelatinolytic activities corresponding to proMMP-2 and the active form of MMP-2 were detected in the wounds 3 and 7 days after surgery. In situ zymography localized gelatinolytic activities at the wound site. These activities were almost completely abolished by an MMP inhibitor, indicating that the gelatinolytic activity belongs to metalloproteinases. Conclusions: MMPs, particularly MMP-2/MT1-MMP, play important roles in the degradation of the extracellular matrix in the wound healing process after glaucoma filtration surgery and may represent an important target for therapeutic intervention after glaucoma filtration surgery. Copyright (C) 2007 S. Karger AG, Basel.