BLOOD COAGULATION & HEMORRHAGIC DISEASE

血液凝固

• 批准号: 3337887
• 负责人: WILLIAM R JR BELL
• 金额: $ 13.98万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-09-30 至 1990-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3337887
• 关键词: blood proteins; calcium; cellular pathology; chromatography; coagulation factor VIII; cortisone; cyclic nucleoside monophosphate; enzyme linked immunosorbent assay; fibrinogen; gel electrophoresis; gene expression; hemorrhagic disorders; liver cells; membrane activity; messenger RNA; peptides; plasminogen activator; prostacyclins; protein C; protein kinase C; radioimmunoassay; radiotracer; spectrometry; tissue /cell culture; vascular endothelium

The overall and long term goals of the proposed research is an increased understanding of the mechanisms involved in fibrinogen biosynthesis. Particular emphasis and interest is directed at the understanding of those mechanisms and components involved in controlling the synthesis of plasma fibrinogen by parenchymal cells of the liver. Our recent studies indicate that rat fibrinogen fragment D90 (FDP-D90) (90,000 daltons) in micro molar concentrations directly and specifically induces the synthesis of fibrinogen by isolated hepatocytes in a chemically-defined, serum-free medium. Particular attention will be directed to identify and define the receptor on the hepatocyte responsible for the binding of FDP-D90. Additional studies will be performed to identify if this peptide is internalized and if the individual genes are stimulated to express increased quantities of mRNA for the Aalpha, B beta and gamma chains respectively by utilization of the appropriate genomic C-DNA probes. We will identify if cyclic nucleotides, cAMP, adenylate cyclase and Interleukin 1, protein kinase c, Ca++, and clamodulin are mediators of this synthetic path. Since it is possible that FDP-D90 may synergistically act with a "cofactor," detailed studies have been proposed to identify, isolate and purify this component and identify its cell of origin. Because we have recently demonstrated that human fibrinogen fragments D94 (94,000 daltons) and D78 (78,000 daltons) induce striking morphologic abnormalities in vascular endothelial cells, we propose continuation of these studies to identify the mechanisms involved. Expansion of the binding studies in progress will include verification of stimulation of cyclic AMP via adenylate cyclase. We will also measure secretion of prostacyclin and tissue plasminogen activator by Factor VIII:Ag and thrombomodulin to learn if these components that modulate thrombogenesis are disturbed by these fragments of fibrinogen. These comprehensive studies will provide new information on how rat fibrinogen FDP-D90 specifically induces the stimulation of the biosynthesis of fibrinogen and how human fibrinogen FDP-D78 and D94 induce damage in vascular endothelial cells.

建议研究的总体和长期目标是 增加对纤维蛋白原相关机制的了解 生物合成 特别强调和兴趣是针对 了解这些机制和组件参与 实质细胞控制血浆纤维蛋白原的合成 肝细胞。 我们最近的研究表明大鼠纤维蛋白原 片段D90（FDP-D90）（90，000道尔顿），微摩尔 浓度直接和特异性地诱导合成 纤维蛋白原的分离肝细胞在一个化学定义， 无血清培养基。 将特别注意 识别和定义肝细胞上负责 FDP-D90的结合。 将进行其他研究， 确定这种肽是否被内化， 被刺激表达增加量的mRNA， A α、B β和γ链， 合适的基因组C-DNA探针 我们将确定是否循环 核苷酸，cAMP，腺苷酸环化酶和白细胞介素1，蛋白质 激酶c、Ca++和clamodulin是这种合成的 路径 由于FDP-D90可能协同作用， 对于“辅因子”，已经提出了详细的研究来识别， 分离和纯化该组分并鉴定其来源细胞。 因为我们最近证明了人类纤维蛋白原 片段D94（94，000道尔顿）和D 78（78，000道尔顿）诱导 血管内皮细胞的显著形态异常， 我们建议继续进行这些研究， 涉及的机制。 正在进行的约束力研究的扩展 将包括通过以下方式验证环AMP的刺激 腺苷酸环化酶 我们还将测量前列环素的分泌 和组织纤溶酶原激活剂通过因子VIII：Ag和 血栓调节蛋白来了解这些调节 血栓形成被这些纤维蛋白原片段干扰。 这些全面的研究将提供新的信息， 大鼠纤维蛋白原FDP-D90特异性诱导刺激 纤维蛋白原的生物合成以及人纤维蛋白原FDP-D 78和 D94诱导血管内皮细胞损伤。