矶松素对中性粒细胞与血小板之间相互作用的影响

Effect of Plumbagin on Platelet-neutrophil Interactions

目的研究矶松素在体内外对血小板聚集功能及中性粒细胞与血小板之间相互作用的影响.方法采用Born方法测定不同状态(非激活或激活态)中性粒细胞对洗涤血小板聚集功能的影响;应用玫瑰花结试验及Born方法分别探讨矶松素对中性粒细胞与血小板间的相互作用.结果在体外,矶松素呈浓度相关性明显抑制ADP、AA及PAF诱导的血小板聚集,其半数抑制浓度(IC50)分别为39.4,82.7和38.1 moL/L;10 mg/kg矶松素灌胃后,其抗血小板聚集作用具体表现为:(1)对ADP引起的血小板聚集:于给药后60 min即显示明显的抑制作用,于120 min达最大效应,至给药后240 min仍具有显著抑制作用;(2)对AA引起的血小板聚集:于灌胃后30 min显效,180 min达最大抑制作用,有效至少可持续240 min;(3)对PAF引起的血小板聚集功能:于给药后90 min起效,药效持续至给药后120 min.矶松素明显降低凝血酶激活的洗涤血小板与中性粒细胞间的粘附率,其IC50为62.9 mol/L,且明显阻抑肉豆蔻佛波醇(fMLP)或血小板活化因子(PAF)激活的中性粒细胞引起的洗涤血小板聚集,其IC50分别为54.3和47.6 mol/L.结论矶松素在体内外均具有明显拮抗ADP、AA或PAF诱导的血小板聚集作用;矶松素显著减少血小板与中性粒细胞间的粘附作用;矶松素明显阻抑激活的中性粒细胞引起的血小板聚集功能.

Objective To investigate the effects of plumbagin on platelet aggregation in vitro and in vivo, and neutrophil-platelet interaction. Methods Born＇s method was used to test the effect of different status of polymorphonuclear leukocytes （PMN） on washed platelet aggregation; the effects of plumbagin on PMN-platelet interaction were observed by use of rosette assay and Born＇s method, respectively. Results The supernatant of non-activated PMN （0.5 ×10^7 cell/mL） significantly inhibited ADP or arachidonic acid （AA） -induced platelet aggregation, and could be enhanced by aspirin; fMLP and platelet-activating factor （PAF） -stimulated PMN suspension or its supernatant could activate platelets. Aspirin, however, had no influence on platelet aggregation induced by fMLP-or PAF-stimulated PMN suspension and supernatant. In vitro, plumbagin markedly inhibited ADP, AA, or PAF-induced platelet aggregation in a concentration dependent manner, with the IC50 value of 39.4, 82.7 and 38.1 mol/L. Intragastric administration, 10 mg/kg of plumbagin showed potent anti-platelet effects： （1） for ADP： significant inhibition 60 min after intragastric administration, showing maximal action at 120 min, and lasting for 240 min; 2） for AA： significant inhibition 30 min after intragastric administration, showing maximal action at 180 min, and lasting for 240 min; （3） for PAF： significant inhibition 90 min after intragastric administration, and lasting for 120 min. Plumbagin significantly suppressed the binding of thrombin-stimulated platelets to PMN, with the IC50 value of 62.9 mol/L, and inhibited fMLP- or PAF-activated PMN-induced platelet aggregation. The IC50 values were 54.3 and 47.6 mol/L, respectively. Conclusions The different conditions of PMN （non-activated or activated） show opposite effects on normal platelets. Briefly, non-activated-PMN inhibit platelet reactivity, whereas activated PMN stimulate it. Plumbagin showe antiplatelet effect induced by ADP, AA or PAF, in vitro or in vivo. Plumbagin significantly inhibit adhesion between PMN and platelets, and markedly suppress platelet aggregation stimulated by activated PMN.