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目的 探讨氯通道在血小板胞浆游离钙和血小板聚集功能调节中的作用。方法 新鲜分离人血小板,以凝血酶为诱导剂,观察氯通道阻断剂DIDS、NFA和钙通道阻断剂SK&F96365、Nife dipine对血小板胞浆游离钙和血小板聚集的单独作用和相互作用。结果 氯通道阻断剂DIDS、NFA可以浓度依赖性地抑制凝血酶 ( 1U/ml)诱导的血小板聚集,对静息血小板胞浆游离钙无明显影响;DIDS、SK&F96365、Nifedipine可以明显降低凝血酶诱导的血小板聚集、钙释放和钙内流,与对照组比较,P<0. 05;DIDS与SK&F96365联合,对凝血酶诱导的血小板聚集、钙释放和钙内流的抑制比各自单独抑制作用明显增高(P<0. 05),两者的作用相互增强;DIDS与Nifedipine联合,对凝血酶诱导的血小板钙释放的抑制比各自单独抑制作用明显增高 (P<0. 05 ),两者可相互增强;NFA与SK&F96365联合,对凝血酶诱导的血小板钙释放的抑制比各自的单独作用明显降低 (P<0. 05 ),两者可相互减弱;NFA与Nifedipine联合,对凝血酶诱导的血小板聚集、钙释放和钙内流的抑制比各自的单独作用明显降低(P<0. 05),两者可相互减弱。结论 氯通道阻断剂DIDS、NFA对人静息血小板胞浆钙浓度无影响;DIDS可抑制凝血酶诱导的血小板聚集、钙释放和钙内流,NFA仅抑制凝血酶诱导的钙释放;氯通道阻断剂和?
Objective To investigate the role of chloride channel in regulating platelet cytoplasmic free calcium and platelet aggregation. Methods Freshly isolated human platelets were treated with thrombin as an inducer. The effects of chloride channel blockers DIDS, NFA and calcium channel blockers SK & F96365 and Nife dipine on platelet cytoplasmic free calcium and platelet aggregation were observed respectively. Results Chloride channel blockers DIDS and NFA could inhibit thrombin (1U / ml) -induced platelet aggregation in a concentration-dependent manner, but had no effect on the free calcium in resting platelets. DIDS, SK & F96365 and Nifedipine could significantly reduce the induction of thrombin Of platelet aggregation, calcium release and calcium influx, compared with the control group, P <0 05; DIDS and SK & F96365 combination of thrombin-induced platelet aggregation, calcium release and calcium influx inhibition than their respective inhibitory effect was significantly higher (P <0.05). The combination of DIDS and Nifedipine significantly inhibited the inhibition of thrombin-induced platelet calcium release compared with that of the control alone (P <0.05), both of which could enhance each other ; NFA combined with SK & F96365, thrombin-induced inhibition of platelet release of calcium than their respective role in significantly reduced (P <0 05), both mutually weakening; NFA and Nifedipine combined thrombin-induced platelet aggregation, The inhibition of calcium release and calcium influx was significantly lower than that of their respective effects (P <0.05), both of which could weaken each other. Conclusions Chloride channel blockers DIDS and NFA have no effect on resting cytoplasmic calcium concentration in human. DIDS can inhibit thrombin-induced platelet aggregation, calcium release and calcium influx. NFA only inhibits thrombin-induced calcium release. Chloride channels Blockers and?