| Project/Area Number |
10670601
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurology
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| Research Institution | Keio University |
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Principal Investigator |
TANAHASHI Norio Dept.of Neurology, School of Medicine, Keio University, Assistant Professor, 医学部, 講師 (10124950)
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| Co-Investigator(Kenkyū-buntansha) |
TOMITA Yutaka Dept.of Neurology, School of Medicine, Keio University, Lecturer, 医学部, 助手 (60276251)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1998: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | endlthelial cell / platelet adhesion / thrombin / GPIIb / IIIa / endo thelial cell / platelet / adhesion |
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| Research Abstract |
We examined whether or not platelets adhere to endothelial cells treated by thrombin in vitro using video enhanced contrast-differential interference contrast (VEC-DIC) microscopy, and also the role of platelet glycoprotein (GP) Iib/IIIa on platelet adhesion to endothelial cells.
METHODS: VEC microscopy: The VEC equipment consisted of an inverted Nomarski microscope and a CCD camera coupled with an image processing processor, monitored at video image magnifications of up to x12000, and recorded on video tape. Experiment 1(N=10): Human aortic endothelial cells (HAEC) were cultured on the coverglass and put in the observation chamber of VEC-DIC microscopy. Human α thrombin 1U/ml with Hank's solution was superfused on HAEC for 30 minutes and washed out. Then, platelet rich plasma (PRP) separated from venous blood of healthy volunteers were superfused with an infusion pump at a low shear rate (10/sec) for 30 min and washed out. Interaction between platelets and endothelial cells was observe
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d by VEC-DIC microscopy and number of platelets adhered to endothelial cells were calculated. Experiment 2 (N=7): Following thrombin pretreatment on HAEC, PRP with TAK029 (1μM, GP Iib/IIIa antagonist) were superfused for 30 min and adhesion of platelets to endothelial cells were observed. Experiment 3 (N=7): As a control, PRP only were superfused for 30 min and adhesion of platelets to endothelial cells were observed. To quantify the degree of platelet adhesion to endothelial cells, number of adhering platelets in a field of 30 micrometer in length and breadth was counted. Consecutive 100 fields was counted and the average number was calculated.
RESULTS: Experiment 1: Platelets adhered to endothelial cells in all experiments and microaggregates of platelets were seen. Average number of platelets adhering and aggragating to endothelial cells were 16.1±9.8/900μmィイD12ィエD1. Experiment 2: Platelets adhesion to endothelial cells were clearly suppressed. Average number of platelets adhering and aggregating to endothelial cells were 0.4±0.3/900μmィイD12ィエD1 (P<0.01, compared to Experiment 1). Experiment 3: Platelets adhesion to endothelial cells were scarecely seen, Average number of platelets adhering and aggregating to endothelial cells were 0.3±0.7/900μmィイD12ィエD1.
CONCLUSION: The above results indicate that platelet adhered to thrombin-treated endothelial cells and GP Iib/IIIa plays an important role for platelets adhesion to endothelial cells in vitro. Less
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