Role of apo (a) in atherogenesis. A study using transgenic rabbits expressing human apo (a)

apo (a) 在动脉粥样硬化形成中的作用。

基本信息

批准号：
10470046
负责人：
WATANABE Teruo
金额：
$ 8.19万
依托单位：
Saga Medical School (2000)University of Tsukuba (1998-1999)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

Transgenic rabbits expressing human apoprotein (a)1. Production of transgenic rabbits :We used liver-specific promoter mouse transferrin promoter and human apo (a) cDNA containing 17 copies of kringle 4. After microinjection of zygotes (2836 in total), we got 96 pups and found that 11 pups had human apo (a) transgene in their genome. After several breeding, we got 3 lines of transgenic rabbits that have detectable apo (a) in the plasma.2. Characterization of transgenic rabbits :Human apo (a) was expressed in the liver as confirmed by Northern blot analysis. Human apo (a) size was about 500kD as determined by SDS-PAGE.In transgenic rabbit plasma, human apo (a) was associated with rabbit apoB to form Lp (a) complex but lipoprotein profile was not effected by apo (a) expression. On a chow diet, transgenic rabbits do not develop any atherosclerotic lesions.3. Characterization of apo (a) transgenic rabbits fed a cholesterol diet for 16 weeks. When on a cholesterol diet, transgenic rabbits developed more atherosclerosis than did nontransgenic control. Increased lesions were found in all arterial trees in transgenic rabbits, including aorta, iliac and carotid arteries, and coronary artery. Immunohistochemical staining study showed that apo (a) may enhance the lesion development by modulating SMC phenotype changes.4. WHHL transgenic rabbits :After cross-breeding between WHHL and transgenic rabbits, we got WHHL transgenic rabbits to investigate whether LDL receptor is involved in Lp (a) catabolism. We found that in the setting of LDL receptor deficiency, Lp (a) was markedly accumulated in the plasma suggesting that Lp (a) clearance is controlled in part, by LDL receptor.

表达人酮蛋白的转基因兔子（a）1。转基因兔的产生：我们使用了肝特异性启动子小鼠转铁蛋白启动子和人apo（a）cDNA，其中包含17份kringle 4。几次繁殖后，我们得到了3行转基因兔子，这些兔子在等离子体中具有可检测的Apo.2。转基因兔子的特征：人apo（a）在肝脏中表达，如北印迹分析所证实。通过SDS-PAGE确定的人apo（A）大小约为500KD。在转基因兔血浆中，人apo（a）与兔APOB相关联，以形成LP（A）复合物，但脂蛋白谱并未受到Apo（A）表达的影响。在食物饮食上，转基因兔子不会产生任何动脉粥样硬化病变3。 Apo（a）转基因兔的表征喂养胆固醇饮食16周。当胆固醇饮食时，转基因兔比非转基因对照更多的动脉粥样硬化。在所有动脉树中发现了病变增加的转基因兔子，包括主动脉，小动脉和颈动脉，以及冠状动脉。免疫组织化学染色研究表明，APO（A）可以通过调节SMC表型的变化来增强病变的发育。4。 WHHL转基因兔子：在WHHL和转基因兔之间杂交后，我们得到了WHHL转基因兔子，以研究LDL受体是否参与LP（A）分解代谢。我们发现，在LDL受体缺乏症的情况下，LP（A）在血浆中显着积累，表明LP（A）清除率部分由LDL受体控制。