Studies on the pathogenesis and pathophysiology of thromboembolisms in the field of surgery. Development of novel techniques for analyzing the regulatory systems of blood coagulation.

外科领域血栓栓塞的发病机制和病理生理学研究。

• 批准号: 63480293
• 负责人: MATSUDA Michio
• 金额: $ 4.22万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63480293/
• 关键词: Molecular abnormality; Abnormal fibrinogen; Protein C; Monoclonal antibodies; Thromboembolic disease; Thrombus formation; 遺伝子解析; アンチトロンビンIII; トロンビン-アンチトロンビンIII複合体; 血管内皮細胞; 血液凝固制御機構

1. Structure analysis of abnormal fibrinogens: We have analyzed 10 abnormal fibrinogens newly referred to us from several institutions an over the country during the term covered by this research grant-in- aid. The mutations identified included two Arg-275 to His; four Arg-275 to Cys; one each of Met-310 to Thr accompanied by N-glycosylated Asn-308; Asp-330 to Tyr and Arg-375 to Gly substitutions. All these mutations could be accounted for by a single base exchange in the codon encoding respective amino acids.This particular region of the gamma chain seems to constitute critical structures required for fibrin polymerization, and their perturbation solely by a single amino acid replacement should have resulted in severely altered fibrin clot formation. Except the gamma Arg-275 to His substitution, all the mutations identified near the carboxy-terminal region of the gamma chain were found to be newly elucidated structural derangements. Gene analysis studies are currently in progress in o … More ur laboratory.In these structure analyses, we successfully applied several monoclonal antibodies recognizing various structures of fibrinogen as will be stated later in item 3.2. Blood coagulation and fibrinolysis which proceed on the cultured human endothelial cells(EC): We have provided lines of evidence that protein C, normally produced by hepatocytes, was also synthesized by the EC's on the basis of identification of protein C molecules as well as messenger RNA for protein C derived from the cultured EC's. This finding has not been reported heretofore, and thus further studies are necessary to more precisely elucidate the EC-mediated regulation of thrombus formation in vivo.3. Monoclonal antibodies raised against substances related to blood coagulation and fibrinolysis: We have prepared and characterized various monoclonal antibodies against fibrinogen, factors IX, X and XIII, plasminogen activator inhibitor and the thrombin-antithrombin complex. An antibody that recognizes the gamma 86-302 residue peptide of fibrinogen and another one that recognizes the NH_2-terminal conformation of plasmic fragment D were successfully utilized for the structure analysis of abnormal fibrinogens. Antibodies raised against other molecules have also been introduced into the analyses of molecular interactions relevant to thrombus formation and its regulation. Less

1.异常纤维蛋白原的结构分析：我们分析了本研究资助期间从全国多个机构新提交的10种异常纤维蛋白原的结构。鉴定的突变包括两个Arg-275至His;四个Arg-275至Cys; Met-310至Thr各一个，伴有N-糖基化Asn-308; Asp-330至Tyr和Arg-375至Gly取代。所有这些突变都可以通过编码相应氨基酸的密码子中的单个碱基交换来解释，γ链的这一特定区域似乎构成了纤维蛋白聚合所需的关键结构，并且仅通过单个氨基酸替换对其的扰动应该会导致严重改变的纤维蛋白凝块形成。除了γ Arg-275替换为His，发现γ链羧基末端区域附近的所有突变都是新阐明的结构紊乱。基因分析研究目前正在进行中， ...更多信息 在这些结构分析中，我们成功地应用了几种识别纤维蛋白原各种结构的单克隆抗体，这将在后面的第3.2条中说明。在培养的人内皮细胞（EC）上进行的血液凝固和纤维蛋白溶解：我们已经提供了一系列证据，表明通常由肝细胞产生的蛋白C也由EC合成，这是基于对来自培养的EC的蛋白C分子以及蛋白C的信使RNA的鉴定。这一发现迄今尚未报道，因此需要进一步的研究来更精确地阐明EC介导的体内血栓形成调节。单克隆抗体提出的物质有关的血液凝固和纤维蛋白溶解：我们已经制备和特点的各种单克隆抗体对纤维蛋白原，因子IX，X和XIII，纤溶酶原激活物抑制剂和凝血酶-抗凝血酶复合物。一种识别纤维蛋白原γ 86-302残基肽的抗体和另一种识别血浆片段D氨基端构象的抗体成功地用于异常纤维蛋白原的结构分析。针对其他分子产生的抗体也被引入到与血栓形成及其调节相关的分子相互作用的分析中。少

项目成果

Yoshihiko URATANI: "Conformation of antithrombin III with defective biological functions derived from a thrombophilic patient." Thrombosis Research. 49. 591-600 (1988)

Yoshihiko URATANI：“源自易血栓患者的具有缺陷生物功能的抗凝血酶 III 构象。”

Nobuhiko YOSHIDA: "Characterization of an apparently lower molecular weight γ-chain variant in fibrinogen Kyoto I.The replacement of γAsn-308 by Lys which caused an accelerated cleavage of fragment D_1 by plasmin and the generation of a new plasmin cleava

Nobuhiko YOSHIDA：“纤维蛋白原京都 I 中分子量明显较低的 γ 链变体的表征。Lys 取代 γAsn-308，导致纤溶酶加速片段 D_1 的裂解，并产生新的纤溶酶裂解

Michio MATSUDA: "Fibrinogen" Excerpta Medica,Amsterdam,New York,Oxford,

松田道雄：《纤维蛋白原》医学摘录，阿姆斯特丹，纽约，牛津，

Takeuchi,Yumiko,: Thrombosis Research,. 51. 117-126 (1988)

竹内由美子，：血栓研究，。

Michio MATSUDA: "A thrombotic state due to an abnormal protein C." New England Journal of Medicine. 319. 1265-1268 (1988)

Michio MATSUDA：“异常蛋白 C 导致的血栓状态。”