Reduced choroidal neovascular membrane formation in matrix metalloproteinase-2-deficient mice
Reduced choroidal neovascular membrane formation in matrix metalloproteinase-2-deficient mice
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DOI：
10.1167/iovs.02-0180
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发表时间：
2003-01-01
影响因子：
4.4
通讯作者：
Kvanta, A
中科院分区：
文献类型：
--
作者：
Berglin, L;Sarman, S;Kvanta, A
目的.研究发现，基质金属蛋白酶（MMP）-2在血管生成中的作用，包括脉络膜新生血管（CNV）。为了进一步研究，本研究观察了MMP-2缺陷小鼠实验性CNV的形成。在野生型和MMP-2缺陷型小鼠中通过氪激光光凝眼底诱导CNV。激光处理后MMP-2 mRNA表达的时间过程通过与反义和正义cRNA探针的原位杂交来确定。免疫组化法检测MMP-2蛋白的分布。治疗后10天，在苏木精-伊红染色的连续切片上评价CNV的程度。CNV病变的最大高度通过数字化组织学图像的图像分析来计算。MMP-2 mRNA在激光治疗后第3天在CNV病变中检测到表达，第5天达到高峰，之后缓慢下降。MMP-2 mRNA表达在膜边缘最高。MMP-2的免疫组化染色证实了MMP-2蛋白在CNV病变中的存在。MMP-2基因敲除小鼠的CNV相对厚度较野生型小鼠减少31%（P = 0.006）。本研究表明，MMP-2的mRNA和蛋白在实验性CNV在小鼠中上调。野生型和MMP-2缺陷小鼠之间CNV膜厚度的显著差异表明MMP-2参与小鼠实验性CNV的形成。这些结果表明，MMPs（包括MMP-2）的药理学靶向作用可能会减少年龄相关性黄斑变性等疾病中CNV的形成。
PURPOSE. Findings in studies have suggested a role for matrix metalloproteinase (MMP)-2 in angiogenesis, including choroidal neovascularization (CNV). To investigate further, the current study was conducted to observe the formation of experimental CNV in MMP-2-deficient mice.METHODS. CNV was induced in wild-type and MMP-2-deficient mice by krypton laser photocoagulation of the fundus. The time-course of expression of MMP-2 mRNA after laser treatment was determined by in situ hybridization with anti-sense and sense cRNA probes. MMP-2 protein distribution was determined by immunohistochemistry. Ten days after treatment, the extent of CNV was evaluated on hematoxylin-eosin stained serial sections. The maximum height of the CNV lesions was calculated by image analysis of digitized histologic images.RESULTS. Expression of MMP-2 mRNA was detected in the CNV lesions at day 3 after laser treatment and peaked at day 5, after which it slowly declined. MMP-2 mRNA expression appeared to be highest at the margins of the membrane. Immunostaining for MMP-2 confirmed the presence of MMP-2 protein in the CNV lesions. The CNV lesions of MMP-2-deficient mice showed that relative thickness was reduced by 31% compared with wild-type mice (P = 0.006).CONCLUSIONS. The present study demonstrated that MMP-2 mRNA and protein are upregulated during experimental CNV in the mouse. The marked difference in thickness of the CNV membrane between wild-type and MMP-2-deficient mice shows that MMP-2 is involved in the formation of experimental CNV in the mouse. These results suggest that pharmacologic targeting of MMPs, including MMP-2, may reduce formation of CNV in conditions such as age-related macular degeneration.