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.

在动脉粥样硬化和许多其他炎症过程中，已经实施了增加的基质金属蛋白酶12（M1VIP-L2）。为了定义MMP-12在Viva中的功能，我们生成了转基因兔子，在巨噬细胞特异性启动子（人类的清道夫接收器启动子）的控制下表达了人类MMP-12基因。通过Southern印迹分析，发现两个转基因创始人兔具有人类MMP-12转基因整合。 HMMP-12 mRNA在腹膜和肺泡巨噬细胞中表达，在转基因兔中富含巨噬细胞的组织中表达。在来自转基因兔子的腹膜和肺泡巨噬细胞的条件培养基中检测到高水平的HMMP-12蛋白。 Zymography表明，巨噬细胞分泌的HMMP-12具有酶促活性对β-蛋白。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。与对照兔子相比，转基因兔中的肉芽肿大小显着增加，组织学检查表明，转基因兔子的肉芽肿富集在与HMMP-L2表达增加有关的巨噬细胞中。我们认为，这种具有HMJVLP-12表达增加的转基因兔模型可能成为对MMP-12在炎症性疾病和癌症侵袭中的进一步机械研究的有用模型，它也是测试IMP-12抑制剂体内作用的理想模型。