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題 名 | von Willebrand Factor is Secreted and Distributed in Two Sides of Thrombin-treated Endothelial Cells in Vitro=凝血酵素刺激後, 體外內皮細胞雙向分泌血小板黏附分子免疫細胞化學研究 |
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作 者 | 王長君; 鄭穹翔; 王寧; | 書刊名 | 醫學研究 |
卷 期 | 14:5 1994.03[民83.03] |
頁 次 | 頁303-316 |
分類號 | 415.132 |
關鍵詞 | 內皮細胞; 血小板黏附分子; 免疫細胞化學; von willebrand factor; Endothelial cell; Immunocytochemistry; |
語 文 | 英文(English) |
中文摘要 | 內皮細胞介於血液及血管組織之間,與血液多種成份及管外組織皆有生理及病理之交互影響,如血栓形成、局部發炎、免疫功能及血管或心肌的舒縮控制等。血小板黏附分子(vWF)分泌方式有經常性的由小泡分泌,或受機械性傷害而產生調節性的由韋氏小體分泌。人類臍帶靜脈血管內皮細胞經體外培養三天,以凝血酵素(1U/ml)處理15分鐘。以免疫酵素吸附技術測得培養液中所含凝血蛋白質之量向管腔方向為控制組之3.8倍,向下方之分泌為控制組之1.4倍。培養於塑膠蓋玻片上之內皮細胞經固定後,以抗凝血蛋白質血清作免疫酵素或免疫膠基金細胞化學染色;或以抗內皮素血清及免疫膠基金標示。控制組無第一抗體、及無凝血素處理者為比較。結果顯示,凝血酵素刺激內皮細胞增加凝血蛋白質之分泌,與生化之檢示結果相吻合。而且經基礎性及調節性二種分泌途徑,由小泡及韋氏小體以至細胞外間質累積之凝血蛋白質,皆顯示極強之免疫標示。是以,凝血酵素可能同時刺激內皮細胞血小板黏附因子之經常性及機械性之雙重分泌。 |
英文摘要 | The secretion of von Willebrand factor (vWF) in both constitutive and regulated pathways mediated through vesicles or Weibel-Palade bodies (WPb) respectively are described in cultured human umbilical vein endothelial cells (HUVEC). Competitive enzyme-linked immunosorbent assay (ELISA) analyses demonstrated that release of vWF into the culture medium and subendothelial matrix from thrombin-treated cells was 3.8 and 1.4 times of that from controls. In order to reveal the vWF secretion from cells after thrombin treatment, the immunoelectron microscopy was applied and the HUVEC cultured on coverslips were treated with thrombin. Cells on coverslips were fixed before immunostained with antibodies conjugated with peroxidase or colloidal gold. Subsequently, cells were flat-embedded in resin. The tangential and cross sections were both prepared. The peroxidase-labeled vWF was localized in the vesicles close to the cell surfaces, in WPb-like vacuoles, and on the extracellular matrix. Protrusions of vWF-positive vesicles into the cell processes, fusion vWF-positive vesicles with cytoplasmic filaments and cell membranes were observed. Accumulation of vWF on the extracellular matrix was detected by immunoperoxidase and immunogold labelings. These results suggested that both the constitutive and regulated pathways of the secretion of vWF from HUVEC treated with thrombin are involved. |
本系統中英文摘要資訊取自各篇刊載內容。