Gene engineering, cell biological aproaches to the mechanisms for early stage of atherosclerosis

基因工程、细胞生物学方法研究动脉粥样硬化早期的机制

• 批准号: 05404039
• 负责人: KITA Toru
• 金额: $ 21.25万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05404039/
• 关键词: endothelial cell; VCAM-1; ICAM-1; HB-EGF; PDGF-A,B chain; lysophosphatidyl-choline; P-selectin; T-lymphocyte; PDGF-A, B鎖; リゾフォスファチジルコリン; P selecTin; 動脈硬化; 泡沫細胞; スカベンジャー受容体; 接着分子; WHHLウサギ; 単球接着分子; マクロファージ; LDL; 内皮細胞; WHHL-ウサギ

Vascular endothelial cells, at early stage of atherosclerosis, plays a pivotal role, expressing endothelial leukocytes adhesion molecules (ICAM-1, VCAM-1) , growth factors (HB-EGF,PDGF-A,B chain) and cytokines in response to various pathophysiological stimuli. We have found that lysophosphatidyl-choline (Lyso-PC) , a prominent phospholipid component of atherogenic lipoproteins, such as oxidized LDL,upregulated differentially VCAM-1 and ICAM-1 expression in various cultured endothelial cells. In addition Lyso-PC has been demonstrated to induce gene expression of potent smooth muscle growth factors such as PDGF-A and B chain, and HB-EGF in cultured human endothelial cells. We are now trying to identify some proteins which are phosphorylated by the stimulation of Lyso-PC and verify the signal transduction pathway for Lyso-PC reactions in vascular endothelial cells. We also examined the expression of adhesion molecules and blood cells (such as monocyte-macrophages, T lymphocytes) in the atherosclerotic lesions in vivo, using WHHL-rabbits and cholesterol fed rabbits by immunological methods. At very early stage, around one P selectin molecules are expressed. At 3 week, we found that activated macrophages and after one more week T lymphocytes are identified at same lesions as adhesion molecules are expressed. Finally we found Lyso-PC could induce some growth factors and cytokines in T lymphocytes. We are now studying the molecular mechanisms of Lyso-PC actions to endothelial cells and T-lymphocytes.

血管内皮细胞在动脉粥样硬化的早期阶段起着关键作用，在各种病理生理刺激下表达内皮细胞粘附分子（ICAM-1、VCAM-1）、生长因子（HB-EGF、PDGF-A、B链）和细胞因子。我们已经发现溶血磷脂酰胆碱（Lyso-PC），致动脉粥样硬化脂蛋白，如氧化低密度脂蛋白，上调差异VCAM-1和ICAM-1的表达在各种培养的内皮细胞的一个突出的磷脂成分。此外，已证明Lyso-PC可诱导培养的人内皮细胞中强效平滑肌生长因子（如PDGF-A和B链以及HB-EGF）的基因表达。目前，我们正试图鉴定一些在Lyso-PC刺激下磷酸化的蛋白质，并验证血管内皮细胞中Lyso-PC反应的信号转导途径。用免疫学方法检测了WHHL-兔和高胆固醇喂养兔动脉粥样硬化病变中粘附分子和血细胞（如单核-巨噬细胞、T淋巴细胞）的表达。在非常早期，大约一个P选择素分子表达。在3周时，我们发现活化的巨噬细胞和一周后的T淋巴细胞在相同的病变中被鉴定为粘附分子的表达。最后发现Lyso-PC可以诱导T淋巴细胞产生一些生长因子和细胞因子。我们现在正在研究Lyso-PC对内皮细胞和T淋巴细胞作用的分子机制。

项目成果

Makoto Tanaka,et al.: "Regulation of apolipoprotein B secretion in hepatocytes from Watanabe heritable hyperlipidemic rabbit,an animal model of familial hypercholesterolemia." Atherosclerosis. in press.

Makoto Tanaka 等人：“渡边遗传性高脂血症兔（一种家族性高胆固醇血症动物模型）肝细胞中载脂蛋白 B 分泌的调节”。

Hiroshi Ochi, et al.: "Elevated levels of cyclic AMP inhibits protein kinase‐C independent mechanisms of endothelial PDGF‐B chain and ICAM‐1 gene induction by lysophosphatidylcholine." Circulation Research. 77. 530-535 (1995)

Hiroshi Ochi 等人：“循环 AMP 水平升高会抑制溶血磷脂酰胆碱诱导内皮 PDGF-B 链和 ICAM-1 基因的蛋白激酶 C 独立机制。” 77. 530–535 (1995)。

Makoto Tanaka,et al.: "Regulation of apolipoprotein B production and secretion in response to the change of intracellular cholesteryl ester contents in rabbit hepatocytes." J.Biol.Chem.268. 12713-12718 (1993)

Makoto Tanaka 等人：“根据兔肝细胞内胆固醇酯含量的变化调节载脂蛋白 B 的产生和分泌。”

Toru Kita, et al.: "Lysophosphatidylcholine induced gene expression of endothelial platelet-derived growth factor-b-chain and intercellular 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-链和细胞间粘附分子-1 的基因表达。”

Kume,N.,et al.: "Lysophosphatidylcholine transcriptionally induces growth factor gene expression in cultured human endothelial cells." J.Clin.Invest.93. 907-911 (1993)

Kume,N.,et al.：“溶血磷脂酰胆碱在培养的人内皮细胞中转录诱导生长因子基因表达。”