Platelet adhesion to human brain microvascular endothelial cells in vitro
| Project/Area Number |
12670617
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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 Keio University, School of Medicine, Assistant professor, 医学部, 講師 (10124950)
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| Co-Investigator(Kenkyū-buntansha) |
SATOH Hideki Keio University, School of Medicine, Instructor, 医学部, 助手 (60296556)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2000: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | platelet adhesion / endothelial cell / Argatroban / Beraprost / GPIIb / IIIb antagonist / beraprost / humanbrain microvascular endothelial cell / thrombin / platelet aggregation |
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| Research Abstract |
We examined whether activated platelets adhere to human Brain microvascular endothelial cells (HBEC) in vitro and whether argatroban (selective thrombin inhibitor) or GPIIb/IIIa antagonist (TAK-029) ameliorates the adhesion of activated platelets to HBEC in vitro using VEC-DIC microscopy. In the control group (N=5), HBECs were cultured on a coverglass and put in the observation chamber of VEC-DIC microscopy. Then, platelet rich plasma (PRP) was superfused with an infusion pump at shear rates (10/sec) for 30 min and platelet adhesion to HBEC was observed. In the ADP group (N=9), PRP with ADP (2μM) was superfused for 30 min and washed out. In the argatroban group (N=9) PRP with ADP (2μM) and argatroan (5μg/ml) was superfused and platelet adhesion to HBEC was observed. In the TAK-029 group (N=9), PRP with ADP (2μM) plus TAK-029 (GPIIb/IIIa antagonist; 1μM) was superfused. In the control group, platelet adhesion to HBEC was rarely seen. In the ADP group, platelets adhered to HBEC in all experiments and microaggregates of platelets were seen. In the argatroban and TAK-029 groups, platelet adhesion to HBEC was clearly suppressed. The average number of platelets adhering and aggregating to HBEC was 0.3 ± 0.6/900 μm^2 in the control group, 25.5 ± 11.3/ 900μm^2 (P<0.01, vs control) in the ADP group, 1.8 ± 1.8/ 900 μm^2 in the argatroban group (p<0.01, vs ADP) and 1.6 ± 1.8/ 900 μm^2. The above results showed argatroban and TAK-029 ameliorated adhesion and aggregated pileup of activated platelets to HBEC at a low-flow state in vitro. The above results showed adhesion of ADP-activated platelets to HBEC under flow in vitro is mediated via GPIIb/IIIa. The results also suggests that thrombin produced by platelet activation makes HBEC procoagulant and is the most likely candidate to subsequently induce platelet adhesion to HBEC.
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Report
(3 results)
Research Products
(10 results)
