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Elucidation of MMP-12 functions in atherosclerosis using transgenic rabbit models

使用转基因兔模型阐明 MMP-12 在动脉粥样硬化中的功能

基本信息

批准号：
13470046
负责人：
WATANABE Teruo
金额：
$ 7.68万
依托单位：
佐賀医科大学
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470046/
关键词：
transgene animal model transgenic rabbit extracellular matrix MMPs atherosclerosis inflammation hyperlipidemia 細胞外質

项目摘要

Increased matrix metalloproteinase-12 (M1VIP-l2) has been implicated in atherosclerosis and many other inflammatory processes. To define MMP-12 functions in viva, we generated transgenic rabbits that expressed human MMP-12 gene under the control of a macrophage-specific promoter, the human scavenger receptor promoter. Two transgenic founder rabbits were found n have human MMP-12 transgene integration by Southern blot analysis. hMMP-12 mRNA was expressed in peritoneal and alveolar macrophages, and in tissues enriched in macrophages in transgenic rabbits. High levels of hMMP-12 protein were detected in the conditioned media of cultured peritoneal and alveolar macrophages from transgenic rabbits. Zymography showed that hMMP-12 secreted from macrophages possessed enzymatic activity toward β-casein. To evaluate the expression of hMMP-12 in inflammatory sites, we used carrageenan-induced granulomas as an in vivo model for tissue macrophages and foam cells. Granuloma size in transgenic rabbits was significantly increased compared to that in control rabbits, and histological examination revealed that granulomas of transgenic rabbits were enriched in macrophages associated with increased hMMP-l2 expression. We believe that this transgenic rabbit model with increased expression of hMJvlP-12 may become a useful model for further mechanistic studies of MMP-12 in inflammatory diseases and cancer invasion, it is also an ideal model for testing the in vivo action of IMP-12 inhibitors.

基质金属蛋白酶-12（M1 VIP-12）的增加与动脉粥样硬化和许多其他炎症过程有关。为了确定MMP-12在体内的功能，我们产生了转基因兔，表达人MMP-12基因的控制下的巨噬细胞特异性启动子，人清道夫受体启动子。Southern杂交检测发现2只转基因母兔中整合了人MMP-12基因。hMMP-12 mRNA在转基因兔的腹腔和肺泡巨噬细胞以及富含巨噬细胞的组织中表达。转基因兔腹腔巨噬细胞和肺泡巨噬细胞条件培养液中检测到高水平的hMMP-12蛋白。酶谱分析表明，巨噬细胞分泌的hMMP-12对β-酪蛋白具有酶促活性。为了评估hMMP-12在炎症部位的表达，我们使用角叉菜胶诱导的肉芽肿作为组织巨噬细胞和泡沫细胞的体内模型。与对照组相比，转基因兔肉芽肿的大小显著增加，组织学检查显示转基因兔肉芽肿中富含巨噬细胞，并与hMMP-12表达增加相关。我们认为，这种hMMP-12表达增加的转基因兔模型可能成为进一步研究MMP-12在炎性疾病和癌症侵袭中的机制的有用模型，它也是测试IMP-12抑制剂体内作用的理想模型。