Development of new drug for intractable hyperlipidemia and its clinical application

顽固性高脂血症新药的研制及其临床应用

• 批准号: 07557073
• 负责人: KITA Toru
• 金额: $ 9.6万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557073/
• 关键词: Cholesterol; Apo B-protein; MTP; VLDL; CHO-cell; Hepatocytes; HSP-70; Transgenic mouse; LDL; LDL受容体; HMG-CoA還元酵素阻害剤; 家族性複合型高脂血症; 家族性高コレステロール血症; WHHLウサギ

We found apolipoprotein B-100 (apo B) secretion is regulated in response to the change of intracellular cholesteryl ester contents by adding low density lipoprotein (LDL). LDL caused a significant dose-dependent increase in apo B secretion, because of preventing the degradation of apo-B protein itself. Immediately after apo B protein is translated from mRNA of apo B,microsomal triglyceride transfer protein (MTP) makes assembly with apo B protein. As a result, VLDL particles are formed. Therefore MTP plays an important role for regulating VLDL production and secretion. To investigate this regulation, we first cloned full length of cDNA for MTP,succeeded to transfect MTP cDNA to CHO cells and detected MTP protein. We are now doing following experiments.1) To establish MTP overexpression system in rabbit hepatocytes.2) To establish transgenic mouse which over express MTP cDNA using transferin promoter.3) To analyze proteins which could regulate post-translational modification of apo-B protein, suchi as MTP,HDP-70 and unknown protein.

我们发现，通过添加低密度脂蛋白（LDL），载脂蛋白 B-100（apo B）的分泌会随着细胞内胆固醇酯含量的变化而受到调节。 LDL 可以防止 apo-B 蛋白本身的降解，导致 apo B 分泌显着呈剂量依赖性增加。当apo B mRNA从apo B mRNA翻译出来后，微粒体甘油三酯转移蛋白(MTP)立即与apo B蛋白组装。结果，形成VLDL颗粒。因此MTP对于调节VLDL的产生和分泌起着重要作用。为了探究这一调控机制，我们首先克隆了MTP的cDNA全长，并成功将MTP cDNA转染至CHO细胞并检测了MTP蛋白。我们目前正在做以下实验：1）建立兔肝细胞MTP过表达系统。2）建立利用转铁蛋白启动子过表达MTP cDNA的转基因小鼠。3）分析调节apo-B蛋白翻译后修饰的蛋白，如MTP、HDP-70和未知蛋白。

项目成果

Circulation Research. (in press).

流通研究。

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Iehara,N.et al.：“小鼠平滑肌表型的分化”。

Kita, T., Tanaka, M., Otani, H., Kume, N.& Yokode, M.: "Lessons concerning human lipoprotein metabolism from the study of the WHHL rabbit." Atherosclerosis. X. 111-113 (1995)

北 T.、田中 M.、大谷 H.、久米 N.

Toru Kita, et al.: "Lysophosphatidylcholine induced gene expression of endothelial platelet-derived growth factor-b-chain and intecellular adhesion molecule-1." Medical Science Symposia Series. (in press).

Toru Kita 等人：“溶血磷脂酰胆碱诱导内皮血小板衍生生长因子-b-链和细胞内粘附分子-1 的基因表达。”

Kita,T.et al.: "Induction of endothelial platelrt-derived frowth gactor-b-chain and intercellular adhesion molecule-1 by lysophosphatidylcholine"

Kita,T.et al.：“溶血磷脂酰胆碱诱导内皮细胞板衍生的泡沫因子-b-链和细胞间粘附分子-1”