流動状態下での血小板血栓形成を連続的に定量化する測定システムの開発

• 研究課題/領域番号: 07557079
• 研究種目: 基盤研究(A)
• 配分区分: 補助金
• 応募区分: 展開研究
• 研究分野: 血液内科学
• 研究機関: 慶応義塾大学
• 研究代表者: 池田 康夫 慶應義塾大学, 医学部, 教授 (00110883)
• 研究分担者: 小田 淳 慶應義塾大学, 医学部, 助手 (50255436) ; 村田 満 慶應義塾大学, 医学部, 講師 (50174305) ; 半田 誠 慶應義塾大学, 医学部, 講師 (40129614)
• 研究期間 (年度): 1995 – 1997
• 研究課題ステータス: 完了 (1997年度)
• 配分額: 13,500千円 (直接経費: 13,500千円); 1997年度: 2,600千円 (直接経費: 2,600千円); 1996年度: 2,900千円 (直接経費: 2,900千円); 1995年度: 8,000千円 (直接経費: 8,000千円)
• キーワード: 血小板粘着 / 血液の速度 / 接着分子 / 血小板膜蛋白 / ヘマトクリット / 抗血小板薬 / ずり速度 / 血栓

研究概要

我々は独自に開発した水晶振動子微量質量センサー(QCM)を組み込んだ流動状態下での簡便かつ定量性のある血小板粘着連続測定装置を用いて生体内での血栓形成機序について種々の検討を加えてきた。この装置は今まで切り離すことが困難であった血小板の粘着(血小板と基質の粘着)とそれに続く血小板同士の凝集反応(血小板と血小板の接着)のうち、初期反応である固相化蛋白への血小板の粘着のみを定量できることが明かとなった。生理的に重要なコラーゲン(CL)ならびにフォン・ビルブランド因子(vWf)への血小板粘着は血流速度依存性(310/秒〜2、400/秒)でかつヘマトクリット依存性(20-45%)あった。循環赤血球量の変化は、血液の流動状態、血中のADP濃度、および壁付近の見かけの血小板数などの因子を介して、流動状態下における血小板粘着量に影響することが明らかとなった。CLへの血小板粘着は、二つのシステム(GPIb/IX-vWf;GPIa/IIa-CL)が互いに独立して存在していることが明らかとなった。すでに臨床的に汎用されている抗血小板剤であるアスピリンやシロスタゾールは両者への血小板粘着に引き続き起こる凝集を阻害するものの、血小板粘着自体はむしろ増加させることが明らかとなった。一方、PGE_1はvWfへの粘着を強力に阻害した。本装置は今まで不明であった血小板の初期の接着反応のみを簡便にかつ定量的に連続測定でき、新しい抗血小板剤の開発等今後の臨床応用に有用であることが示唆された。

研究成果

• [文献書誌] Kameda H, Morita I, Handa M., Ikeda Yetal: "Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin." Brit J Haematol 97. 97. 348-355 (1997.)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Goto S., Sakai H., Ikeda Y, and Handa S.:"Acute myocardial infarction plasma augments platelet thrombus growth in high shear rates." The Lancet. 349(9051). 543-544 (1997)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Yokoyama K., Handa M., and Ikeda Y.et al: "Characterization of the novel murine monoclonal anti-von Willebrand factor(vWf)antibody GUR76-23 which inhibits vWf interaction with α IIb β 3 but not αvβ3 integrin." Biochem Biophys Res Commun. 234. 147-152 (1997)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Iijima K, Murata M., Handa M., Ikeda Y.et al: "High shear stress attenuates agonist-induced,glycoprotein IIb/IIIa-mediated platelet aggregation when von Willebrand factor binding to glycoprotein Ib/IX is blocked." Biochem Biophys Res Commun. 233(3). 796-800 (1997)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Miyakawa Y., Oda A., Handa M., Ikeda Y.et al: "Thrombopoietin and thrombin induce tyrosine phosphorylation of vav in human blood platelets." Blood. 89(8). 2789-2798 (1997)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Ikeda Y., Murata M., and Goto S.: "von Willebrand factor-dependent shear-induced platelet aggregation:basic mechanisms and clinical implications." The Annals of the New York Academy of Sciences. 811. 325-336 (1997)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Moriki T., Murata M., Handa M., Ikeda Y.: "Expression and functional characterization of an abnormal platelet membrane glycoprotein Ib α(Met239→Val)reported in patients with platelet-type von Willebrand disease." Blood. 89(10). 3565-3573 (1997)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Kameda H., Morita I., Handa M., Ikeda Y et al: "Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin." Brit J Haematol. 97. 348-355 (1997)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Goto S., Sakai H., Ikeda Y,and Handa S.: "Acute myocardial infarction plasma augments platelet thrombus growth in high shear rates." The Lancet. 349(9051). 543-544 (1997)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Yokoyama K., Handa M., Oda A.and Ikeda Y.et al: "Characterization of the novel murine monoclonal anti-von Willebrand factor (vWf) antibody GUR76-23 which inhibits vWf interaction with alpha IIb beta 3 but not alpha v beta 3 integrin." Biochem Biophys Res Commun. 234. 147-152 (1997)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Iijima K., Murata M., Handa M., Ikeda Y.et al: "High shear stress attenuates agonist-induced, glycoprotein IIb/IIIamediated platelet aggregation when von Willebrand factor binding to glycoprotein Ib/IX is blocked." Biochem Biophys Res Commun. 233(3). 796-800 (1997)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Miyakawa Y., Oda A., Handa M., Ikeda Y.et al: "Thrombopoietin and thrombin induce tyrosine phosphorylation of vav in human blood platelets." Blood. 89(8). 2789-2798 (1997)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Ikeda Y., Murata M., and Goto S.: "von Willebrand factor-dependent shear-induced platelet aggregation : basic mechanisms and clinical implications." The Annals of the New York Academy of Sciences. 811. 325-336 (1997)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Moriki T., Murata M., Handa M., Ikeda Y.: "Expression and functional characterization of an abnormal platelet membrane glycoprotein Ib alpha (Met239*Val) reported in patients with platelet-type von Willebrand disease." Blood. 89(10). 3565-3573 (1997)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Kameda H.,Morita I.,Handa M.,Ikeda Y et al: "Re-expression of functional P-selectin molecules on the endothelial cell surface by repeated stimulation with thrombin." Brit J Haematol 97. 97. 348-355 (1997)
• [文献書誌] Goto S.,Sakai H.,Ikeda Y,and Handa S.: "Acute myocardial infarction plasma augments platelet thrombus growth in high shear rates." The Lancet. 349(9051). 543-544 (1997)
• [文献書誌] Yokoyama K.,Handa M.,and Ideda Y.et al: "Characterization of the novel murine monoclonal anti-von Willebrand factor (vWf) antibody GUR76-23 which inhibits vWf interaction with α IIb β 3 but not α v β 3 integrin." Biochem Biophys Res Commun. 234. 147-152 (1997)
• [文献書誌] Iijima K,Murata M.,Handa M.,Ikeda Y.et al: "High shear stress attenuates agonist-induced,glycoprotein IIb/IIIa-mediated platelet aggregation when von Willebrand factor binding to glycoprotein Ib/IX is blocked." Biochem Biophys Res Commun. 233(3). 796-800 (1997)
• [文献書誌] Miyakawa Y.,Handa M.,Ikeda Y.et al: "Thrombopoietin and thrombin induce tyrosine phosphorylation of vav in human blood platelets." Blood. 89(8). 2789-2798 (1997)
• [文献書誌] Ikeda Y.,Murata M.,and Goto S.: "von Willebrand factor-dependent shear-induced platelet aggregation : basic mechanisms and clinical implications." The Annals of the New York Academy of Sciences. 811. 325-336 (1997)
• [文献書誌] Moriki T.,Murata M.,Handa M.,Ikeda Y.: "Expression and functional characterization of an abnormal platelet membrane glycoprotein Ib α (Met239→Val) reported in patients with platelet-type von Willebrand disease." Blood. 89(10). 3565-3573 (1997)
• [文献書誌] Kitaguchi T: "Establishment and characterization of transgenic mice expressing human platelet glycoprotein lba" 220. 418-424 (1996)
• [文献書誌] Oda,A: "Thrombopoietin primes human platelet aggregation induced by shear stress and by multiple agonists" Blood. 87. 4664-4670 (1996)
• [文献書誌] Oda,A: "_p120^<c-cbl> is present in human blood platelets and is differenrially involned in signaling by thrombopoietin and thrombin" Blood. 88. 1330-1338 (1996)
• [文献書誌] Shiraga,M: "Affinity modulation of the platelet integrin αIIbβ3 by α-chymotrypsin:a possible role for Na^+/Ca^<2+> exchanger" Blood. 88. 2594-2602 (1996)
• [文献書誌] Tokuhira M: "A novel regulatory epitope difined by a murine monoclonal antibody to the platelet GPIIB-IIIa complex (αIIbβ3 integrin)" Thromb Haemost. 76. 1038-1046 (1996)
• [文献書誌] Oda,A: "CrKl is constitutively tyrosine phosphorylated in platelets from chronic myelogenous leukemid patients and inducibly phosphorylated in normal platelets stimulated by thrombopoietin" Blood. 88. 4304-4313 (1996)
• [文献書誌] Goto,S: "Synergistic effect of epinephrine and shearing on platelet activation" Throm Res. 84. 351-359 (1996)
• [文献書誌] Sakamaki F.: "Plasmasoluble form of P-Selectin is eleuated in acute lung in jury." Am.J.Respirat & Crinical Care Med.151. 1824-1826 (1995)
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• [文献書誌] Takahashi H.: "Substitution of Vol for Met at residue 239 of platelet glycoprotein Ib in Japanese patients with plotelet-type von Willebrand disease." Blood. 85. 727-733 (1995)
• [文献書誌] Sakamaki F.: "Soluble form of P-Selectin in Plasma is elevated in acute 1 ung injury." Am.J.Resp.Crit.Care Med.151. 1821-1826 (1995)
• [文献書誌] Fujimura Y.: "Isolation and characterization of jararaca GPIb BP, asnake venom antagonist Specific to platelet glycoprotein Ib." Thromb,and Haemost.74. 743-750 (1995)
• [文献書誌] Goto S.: "Cnanacterization of the unigue mechanism mediating the Shear-dependent binding of Soluble von Willebrand factor to platelets." J.Biol.Chem.270. 23352-23361 (1995)
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• [文献書誌] Miyakawa Y.: "Recombinant thrompopotelin induces rapiaprotein throsine phosphory latiom of janus kinase 2 and shcin human blood platelet." Blood. 86. 23-27 (1995)
• [文献書誌] Oda,A.: "Pnotin iyhe sine phos phory lation in hum an plate lets during Shear-Stress-induced. Platelet aggregation (SIPA) is reguratea by glycopratein (GP) Ib/Ixas well as GIIb/IIIa and requies intact cytoskeleton and endogenepus A bp." Thromb.Haemost.74. 736-742 (1995)