Etiology and pathophysiology of thrombosis : A molecular biological aproach to elucidate disturbed mechanisms of blood coagulation and its inhibition.

血栓形成的病因学和病理生理学：一种分子生物学方法，用于阐明凝血及其抑制的紊乱机制。

• 批准号: 08407034
• 负责人: MATSUDA Michio
• 金额: $ 15.23万
• 依托单位: Jichi Medical School, School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08407034/
• 关键词: hereditary dysfibrinogen; thromboembolism; electron microscopy; fibrin; adhesion molecule; beta1-integrin; kininogen; キニノゲン; フィブリノゲン; 血栓症; 出血性素因; インテグリン; 腫瘍の転移; 分子異常; フィブリンの電顕像; フィブリン重合; 一過性虚血発作; 接着分子受容体; RGD配列

I.Analyses of hereditary dysfibnnogens : During the three year-term of studies supported by the Grant-in-Aid for Scientific Research, No. 08407034, we were able to analyze more than 10 abnormal fibrinogen molecules, some of which had been associated with clinical manifestations of either bleeding or thrombosis, or both. Fibrinogen (Fbg) Caracas II with a unique Aalpha Ser-434 to Asn substitution, to which an extra oligosaccharide is linked (J Biol Chem 266 : 11575-11581, 1991) was further studied by electron microscopy (EM). The Caracas II fibrin fibers are found to be loosely associated and fibrin gels appeared to consist of irregularly aligned fibrin bundles with scattering large caves and pores. The permeability experiment showed a high permeability rate as compared with normal fibrin gels (J Biol Chem 271 : 4946-4953, 1996). On the other hand, Fbg Marburg (truncation of 150 amino acids in the Aalpha-chain and partly linked with serum albumin) associated with severe postoperative bl … More eeding accompanied by successive recurrent thrombo-embolic complications was found to partly linked with one or two serum albumin molecules by EM as anticipated by SDS-PAGE.The fibrin gels are composed of extremely thin and highly branched fibrin fibers, forming extraordinarily compact gels. These fibrin clots account for thrombo-embolic complications together with bleeding due to delayed fibrin clot formation. Part of these data was published in BLOOD (91 : 3282-3288,1998), and the remainder is now in preparation for publication. Four other abnormal Fbg's were also characterized and published (Kurashiki : gamma Gly-268 to Glu, Blood 87 : 4686-4694, 1996 ; Kumamoto : Aalpha Arg-19 to Gly, Jpn J Thromb Hemost 8 : 382-392, 1997 ; Kamogawa : gammaArg-275 to Ser, Thromb Haemostas, in press ; Niigata : Bbeta Asn-160 to Ser with an extra oligosaccharide N-linked to Bbeta Asn-158 ; Blood, in press). In three other dysfibrinogens, we have also identified new types of point mutations, Fbg's Pretoria (gamma Cys-139 to Tyr) ; Tokyo V (gammaAla-327 to Thr) and Osaka VI (12 amino acid extension due to the stop codon TAA to AAA for Lys). Further analyses on these molecules are going on, and will be submitted for publication.II.Molecular mechanisms of fibrin (Fbn) to function as adhesion molecule : Fbg functions as adhesion molecule only after transition to fibrin. When human fibroblasts were cultured on a fibrin monolayer, the bi-integrin as well as the already known beta3-integrin was expressed on the cell surface, and spreading of cells progressed in an RGD-dependent manner (J Biol Chem 272 : 8824-8829, 1997). When human glioma cells were cultured, a two-step mode of spreading via the beta1 -integrin was noted : firstly with fibrin (Aalpha RGD 572-574) and then with the autologous fibronectin secreted and incorporated in the extra-cellular matrix. These findings are now in the status of"in press" in Thrombosis Research. Furthermore, plasma kininogen was found to behave not only as an adhesion molecule but also as an inhibitor. These opposing effects were characterized and published (J Biochem 124 : 473-484, 1998). Less

遗传性不适性的i.analyses：在由科学研究授予的三年期间的研究中，第08407034号，我们能够分析10个以上异常的纤维蛋白原分子，其中一些分子与出血或肿瘤临床相关。电子显微镜（EM）进一步研究了具有独特的Aalpha Ser-434 to ASN取代的独特Aalpha Ser-434 to ASN取代的Caracas II（J Biol Chem 266：11575-11581，1991）。发现加拉加斯II纤维蛋白纤维松散相关，纤维蛋白凝胶似乎由不规则排列的纤维蛋白束和散射大洞和毛孔组成。与正常纤维蛋白凝胶相比，渗透率实验显示出高渗透率（J Biol Chem 271：4946-4953，1996）。 On the other hand, Fbg Marburg (truncation of 150 amino acids in the Aalpha-chain and partly linked with serum albumin) associated with severe posterior BL … More eding accumulated by successful recurrent thrombo-embolic complications was found to partly linked with one or two serum albumin molecules by EM as anticipated by SDS-PAGE.The fibrin gels are composed of extremely thin and highly branched fibrin纤维，形成非常紧凑的凝胶。这些纤维蛋白布解释了血栓栓塞并发症，以及由于纤维蛋白凝块的延迟形成而导致的出血。这些数据的一部分发表在血液中（91：3282-3288,1998），其余的现在正在准备出版。还表征和发表了其他四个异常FBG（Kurashiki：Gamma Gly-268 to Glu，血液87：4686-4694，1996; Kumamoto：aalpha arg-19 to Gly，gly，jpn j Phlomb Hemost 8：382-392，1997; kamogawa; niigata：BBETA ASN-160与BBETA ASN-158的额外的寡糖N-链接；在其他三种动物纤维纤维蛋白中，我们还鉴定了新型点突变，即FBG的Pretoria（伽玛Cys-139 to Tyr）；东京V（Gammaala-327至THR）和大阪VI（由于停止密码子TAA到AAA而引起的12个氨基酸延伸）。对这些分子的进一步分析正在进行中，并将提交出版。II。纤维蛋白（FBN）的分子机制（FBN）充当粘合分子：FBG仅在过渡到纤维蛋白后仅充当粘合分子。当在纤维蛋白单层上培养人成纤维细胞时，双积整合蛋白以及已知的β3-整合蛋白在细胞表面表达，并以RGD依赖性方式进行细胞的扩散（J Biol Chem 272：8824-8829，1997）。当培养人神经胶质瘤细胞时，通过Beta1-整合蛋白扩散了两步的模式：首先，使用纤维蛋白（Aalpha RGD 572-574），然后与自体纤连蛋白纤维蛋白分泌，并掺入外细胞基质中。这些发现现在处于血栓形成研究中的“印刷”状态。此外，发现血浆差异不仅作为粘合分子，而且作为抑制剂的行为。这些相反的影响的特征和发表（J Biochem 124：473-484，1998）。较少的

项目成果

MUTO, Terukazu: "Factor XIII supplement therapy-effects on disturbances of wound healing." Med.Progr.10. 16-19 (1997)

MUTO、Terukazu：“因子 XIII 补充疗法 - 对伤口愈合障碍的影响。”

松田道生: "フィブリノゲンの誘導体,とくに可溶性フィブリン(soluble fibrin)とDダイマー(D dimer)について : その生成と存在様式についての考察.その1.可溶性フィブリン" 日本血栓止血学会誌. 8. 24-32 (1997)

Michio Matsuda：“关于纤维蛋白原衍生物，特别是可溶性纤维蛋白和 D 二聚体：对其形成和存在方式的考虑。第 1 部分。可溶性纤维蛋白”日本血栓和止血学会杂志 8. 24-32 (1997)。

SUGO, Teruko: "Factor XIIIa-cross-linking of the Marburg Fibrin : Formation of αm・γ n-heteromultimers and the α-chain-linked albumin・γ complex, and disturbed protofibril assembly resulting in acquisition of plasmin-resistance relevant to thrombophilia." B

SUGO，Teruko：“马尔堡纤维蛋白的 XIIIa 因子交联：αm·γ n-异多聚体和 α-链连接的白蛋白·γ 复合物的形成，以及原纤维组装的紊乱，导致获得与血栓形成倾向相关的纤溶酶抗性.“B

Funayama H,Sakata Y,Kitagawa S,Ikeda Y,Takahashi M,Masuyama J,Mimuro J,Matsuda M,Shimada K: "Monocytes modulates the fibrinolytic balance of endothelial cells." Thromb Res. 85. 377-385 (1997)

Funayama H，Sakata Y，Kitakawa S，Ikeda Y，Takahashi M，Masuyama J，Mimuro J，Matsuda M，Shimada K：“单核细胞调节内皮细胞的纤溶平衡。”

Shinji Asakura: "Fibroblats spread on immobilized fibrin monomer by mpbilizing a β1-class integrin,together with a vitronectin receptors,αVβ3 on their surface." J.Biol.Chem.,in press. (1997)

Shinji Asakura：“成纤维细胞通过在其表面固定 β1 类整合素以及玻连蛋白受体 αVβ3 来在固定的纤维蛋白单体上扩散。”（J.Biol.Chem.）（1997 年）。