An Investigation of the Expression of Thrombomodulin in Cultured Human Umbilical Vein Endothelial Cells

培养人脐静脉内皮细胞中血栓调节蛋白的表达研究

• 批准号: 02454519
• 负责人: MARUYAMA Ikuro
• 金额: $ 1.41万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454519/
• 关键词: Thrombomodulin; endothelial cells; cyclic AMP; cytokine; thrombomodulin; TNF; ILーI; 動脈硬化; 血栓症; テラトカルチノ-マ

We investigated the expression of thrombomodulin in cultured human umbilical vein endothelial cells(HUVEC) and human megakaryoblastic leukemia cell line, MEG-01 and got the following results.1) An incubation of the cell swith 3-5 mM dbcAMP upregulated the expression of cellsurface throbomodulin. The cell-surface TM was increased by aproximately by 250 % in HUVEC and by 600 % in MEG-01.2) The other agents such as IBMX, forskolin, prostacyclin, which increase in intracellular cAMP, also increase in the cell-surface thrombomodulin.3) IL-1 and TNF alpha, however, decreased in the cell-surface thrombomodulin by about 50 %.4) We could not find so called cAMP responsive element in the upstream of thrombomodulin genome.5) Two type of thrombomodulin is expressed in CHO cells which constructed by the transfection of human thrombomodulin cDNA. A thrombomodulin with a molecular weight of 75, 000 dalton has been recognized as thrombomodulin. Another thrombomdulin with the 170, 000 dalton is expressed and designated as type 2 thrombomodulin. CHO cells expressed both type of thrombomodulin by equal amount.6) The expression ratio of two type of thrombomodulin was examined by immunoblotting. HUVEC in normal culture expressed type two thrombomodulin about 5 %.

我们研究了培养的人脐静脉内皮细胞(HUVEC)和人巨核细胞白血病细胞系MEG-01中血栓调节蛋白的表达，得到以下结果：1)细胞与3-5 mM dbcAMP一起孵育可上调细胞表面血栓调节蛋白的表达。 HUVEC 中的细胞表面 TM 增加了约 250 %，MEG-01 中增加了 600 %。2) 其他药物，如 IBMX、毛喉素、前列环素，可增加细胞内 cAMP，也增加细胞表面血栓调节蛋白。3) 然而，IL-1 和 TNF α 使细胞表面血栓调节蛋白减少约 50 %.4)我们在血栓调节蛋白基因组上游找不到所谓的cAMP反应元件。5)转染人血栓调节蛋白cDNA构建的CHO细胞中表达两种类型的血栓调节蛋白。分子量为 75, 000 道尔顿的血栓调节蛋白已被公认为血栓调节蛋白。另一种具有 170, 000 道尔顿的血栓调节蛋白被表达并指定为 2 型血栓调节蛋白。 CHO细胞等量表达两种类型的血栓调节蛋白。6)采用免疫印迹法检测两种类型血栓调节蛋白的表达比例。正常培养物中的 HUVEC 表达约 5% 的二型血栓调节蛋白。

项目成果

Ikuro Maruyama,et al: "Expression of Two type of Thrombomodulin in the Cultured Human Umbilical Vein Endothelial Cells and other Cell types." Thromb.Haemast.

Ikuro Maruyama 等人：“两种类型的血栓调节蛋白在培养的人脐静脉内皮细胞和其他细胞类型中的表达”。

Ikuro Maruyama: "Upregulation of Thrombomodulin on cultured Endothelial Cells by cAMP." Annals of the New York Academy of Sciences. 598. 538-539 (1990)

Ikuro Maruyama：“cAMP 上调培养内皮细胞上的血栓调节蛋白。”

Akitada Ichinose,et al: "Two types of abnormal genes for plasminogen in families with a predisposition for thrombosis" Proc.Natl Acad.Sci.88. 115-119 (1991)

Akitada Ichinose 等人：“具有血栓形成倾向的家庭中纤溶酶原的两种异常基因”Proc.Natl Acad.Sci.88。

Akitada Ichinose: "Two types of abnormal genes for plasminogen in families with a predisposition for thrombosis." Proc.Natl.Acad.Sci.USA. 88. 115-119 (1991)

Akitada Ichinose：“有血栓形成倾向的家族中存在两种类型的纤溶酶原异常基因。”

丸山 征郎他: "「血液疾患のバイオテラピ-」(柴田,青木編)リコンビナント トロンボモジュリン" 医歯薬出版, 220 (1991)

Seiro Maruyama 等人：“‘血液疾病生物疗法’（Shibata、Aoki 编辑）重组血栓调节蛋白”Ishiyaku Publishing，220（1991）