1990 Fiscal Year Annual Research Report

tーPAとPAインヒビタ-の結合様式の解明とそれに及ぼすフィブリンの影響の研究

Research Project

Project/Area Number	01480114
Research Institution	Hamamatsu University School of Medicine
Principal Investigator	高田明和浜松医科大学, 医学部, 教授 (80092980)
Co-Investigator(Kenkyū-buntansha)	高田由美子浜松医科大学, 医学部, 助手 (90092981)
Keywords	組織プラスミノ-ゲンアクチベ-タ- / プラスミノ-ゲンアクチベ-タ-インヒビタ- / HUVEC / tーPA / PAIーl / ユ-グロブリン溶解時間
Research Abstract	1 ヒト胎盤のPAIーlのmRNAよりcDNAをとり、これをλgtllのファ-ジに入れてexpressionし、ライブラリ-を作る。抗PAIーl IgGを用いて、免疫学的にヒトPAIーlのcDNAをとる(Nyの方法による)。クロ-ニングの後にPAIーlをE.coli lysogenic strainにexpressさせ、recombinantのPAIーlをとった。これをrPAIーlと名付ける。 2 臍帯静脈の内皮細胞(HUVEC)を培養してtーPA、PAIーlをとることに成功した。cPAIーlと名付ける。 3 rPAIーlは他家の報告の通り不活性であった。これを7Mのグアニジンや10%SDSを加えると活性型になる。これ以外にrPAIーlはプラスミンで一部活性型になることを見いだした。つまりrPAIーlは限定分解により構造をかえてtーPAとの結合能を得ることがわかった(T.Urano et al.Eur.J.Biochem.in press)。 4 HUVECよりの上清にはtーPAとcPAIーlが存在している。つまりcPAIーlも不活型で、これも高濃度のSDS、グアニジン等で活性化される。このcPAIーlにfreshなtーPAを加え、2日間インキュベ-トすると、tーPAはPAIーlと反応して結合することが出来るようになる。我々はこの機作を検討中である。 5 tーPAは血漿中ではPAIーlのみでなく _<α2>AP、ClINH、 _<α2>M等のインヒビタ-と結合している。従ってtーPAの抗原量とユ-グロブリン溶解時間(ELT)の相関をとるとELTは正の相関を示す。しかしtーPAが多くなれば線溶活性は亢進する筈なので負の相関を示さなくてはおかしい。このことは抗原量として測定したtーPAはfreeのtーPA量を反映していないことを意味する。我々はfreeのtーPA量のみに反応するモノクロナル抗体の作製を試み、よい結果を得ている。一方化学反応速度論より計算したfreeのtーPAはELTと負の相関を示し、freeのtーPAとtーPA抗原とは相関しないことがわかった。

Research Products
(17 results)

All Other

All Publications (17 results)

[Publications] Akikazu TAKADA: "Fluctuation of euglobulin lysis time,tissue plasminogen activator,and free and total plasminogen activator ingibitor levels in plasma in daytime" Thrombosis Research. 57. 13-20 (1990)
[Publications] Kenji SAKAKIBARA: "Significance of urinary complement components in various glomerular diseases" Thrombosis Research. 57. 625-637 (1990)
[Publications] Tetsumei URANO: "Relationship between euglobulin clot lysis time and the plasma levels of tissue plasminogen activator and plasminogen activator inhibitor l" Thrombosis and Haemostasis. 63. 82-86 (1990)
[Publications] Akikazu TAKADA: "Prevention of severe bleeding by tranexamic acid in a patient with disseminated intravascular coagulation" Thrombosis Reserach. 58. 101-108 (1990)
[Publications] Kaoru SAITO: "The concentration of tissue plasminogen activator and urokinase in plasma and tissues of patients with ovarian and uterine tumors" Thrombosis Research. 58. 355-366 (1990)
[Publications] Andzrej RYDZEWSKI: "Plasminogen activators and plasminogen activator inhibitor (PAIーl) in type II diabetes mellitus" Fibrinolysis. 4. 183-188 (1990)
[Publications] Akikazu TAKADA: "Changes in plasma levels of tーPA,PAIーl and tーPAーPAIーl complex in patients with cerebrovascular thrombosis infused with tーPA" Thrombosis Reserch. 59. 375-381 (1990)
[Publications] Kazuyuki SAKATA: "Clinical significance of plasminogen activator inhibitor activity in patients with exerciseーinduced ischemia" American Heart Journal. 120. 831-838 (1990)
[Publications] Andzrej RYDZEWSKI: "Changes in plasminogen activator inhibitor l and tiisue type plasminogen activator during exercise in patients with coronary artery disease" Haemostasis. 20. 305-312 (1990)
[Publications] Tetsumei URANO: "Fluctuation of tーPA and PAIーl antigens levlels in plasma:Differnce of their fluctuation patterns between male and female" Thrombosis Research. 60. 55-62 (1990)
[Publications] Michal H.PIETRASZEK: "Enhanced platelet response to serotonin in Buerger's disease" Thrombosis Research. 60. 241-246 (1990)
[Publications] Michal H.PIETRASZEK: "Circadian rhythm of serotonin; Influence of age." Thrombosis Research. 60. 253-257 (1990)
[Publications] Tetsumei URANO: "Changes of parameters in fibrinolytic system caused by mental stress" Thrombosis Research. 60. 501-507 (1990)
[Publications] Tetsumei URANO: "Fibrinogen 4ーCurrent basic and clinical aspects" Elsevier Science Publishers B.V.The Netherlands, 224 (1990)
[Publications] Akikazu TAKADA: "Fibrinogen 4ーCurrent basic and clinical aspects" Elsevier Science Publishers B.V.The Netherlands, 224 (1990)
[Publications] Akikazu TAKADA: "Protease Inhibitors" Elsevier Science Publishers B.V.The Netherlands, 233 (1990)
[Publications] Ymiko TAKADA: "Protease Inhibitors" Elsevier Science Publishers B.V.The Nether lands, 233 (1990)

1990 Fiscal Year Annual Research Report

tーPAとPAインヒビタ-の結合様式の解明とそれに及ぼすフィブリンの影響の研究

Principal Investigator

高田 明和 浜松医科大学, 医学部, 教授 (80092980)

Research Products

[Publications] Akikazu TAKADA: "Fluctuation of euglobulin lysis time,tissue plasminogen activator,and free and total plasminogen activator ingibitor levels in plasma in daytime" Thrombosis Research. 57. 13-20 (1990)

[Publications] Kenji SAKAKIBARA: "Significance of urinary complement components in various glomerular diseases" Thrombosis Research. 57. 625-637 (1990)

[Publications] Tetsumei URANO: "Relationship between euglobulin clot lysis time and the plasma levels of tissue plasminogen activator and plasminogen activator inhibitor l" Thrombosis and Haemostasis. 63. 82-86 (1990)

[Publications] Akikazu TAKADA: "Prevention of severe bleeding by tranexamic acid in a patient with disseminated intravascular coagulation" Thrombosis Reserach. 58. 101-108 (1990)

[Publications] Kaoru SAITO: "The concentration of tissue plasminogen activator and urokinase in plasma and tissues of patients with ovarian and uterine tumors" Thrombosis Research. 58. 355-366 (1990)

[Publications] Andzrej RYDZEWSKI: "Plasminogen activators and plasminogen activator inhibitor (PAIーl) in type II diabetes mellitus" Fibrinolysis. 4. 183-188 (1990)

[Publications] Akikazu TAKADA: "Changes in plasma levels of tーPA,PAIーl and tーPAーPAIーl complex in patients with cerebrovascular thrombosis infused with tーPA" Thrombosis Reserch. 59. 375-381 (1990)

[Publications] Kazuyuki SAKATA: "Clinical significance of plasminogen activator inhibitor activity in patients with exerciseーinduced ischemia" American Heart Journal. 120. 831-838 (1990)

[Publications] Andzrej RYDZEWSKI: "Changes in plasminogen activator inhibitor l and tiisue type plasminogen activator during exercise in patients with coronary artery disease" Haemostasis. 20. 305-312 (1990)

[Publications] Tetsumei URANO: "Fluctuation of tーPA and PAIーl antigens levlels in plasma:Differnce of their fluctuation patterns between male and female" Thrombosis Research. 60. 55-62 (1990)

[Publications] Michal H.PIETRASZEK: "Enhanced platelet response to serotonin in Buerger's disease" Thrombosis Research. 60. 241-246 (1990)

[Publications] Michal H.PIETRASZEK: "Circadian rhythm of serotonin; Influence of age." Thrombosis Research. 60. 253-257 (1990)

[Publications] Tetsumei URANO: "Changes of parameters in fibrinolytic system caused by mental stress" Thrombosis Research. 60. 501-507 (1990)

[Publications] Tetsumei URANO: "Fibrinogen 4ーCurrent basic and clinical aspects" Elsevier Science Publishers B.V.The Netherlands, 224 (1990)

[Publications] Akikazu TAKADA: "Fibrinogen 4ーCurrent basic and clinical aspects" Elsevier Science Publishers B.V.The Netherlands, 224 (1990)

[Publications] Akikazu TAKADA: "Protease Inhibitors" Elsevier Science Publishers B.V.The Netherlands, 233 (1990)

[Publications] Ymiko TAKADA: "Protease Inhibitors" Elsevier Science Publishers B.V.The Nether lands, 233 (1990)

高田明和浜松医科大学, 医学部, 教授 (80092980)