Inhibitory effect of disodium quercetin-7, 4'-disulfate on aggregation of pig platelets induced by thrombin and its mechanism^1  

槲皮素-7,4’-二硫酸酯对凝血酶诱导猪血小板聚集的抑制作用及其机制(英文)

作　　者：刘文[1] 梁念慈[1] 

机构地区：[1]广东医学院生物化学和分子生物学研究所,湛江中国524023

出　　处：《Acta Pharmacologica Sinica》2000年第8期68-72,共5页中国药理学报（英文版）

基　　金：Project supported by Guangdong Research Foundation for Key Field, № 9306

摘　　要：PL dependent PKC activity, and actin polymerization.To study the inhibitory effect of semi-synthesized quercetin derivatives-disodium quercetin-7,4'-disulfate (DQD) on the platelet aggregation induced by thrombin and its mechanism. METHODS: Platelet aggregation was analysed by turbidimetry. Cytosolic free calcium concentration ([Ca2+]i) was determined by Fura-2 fluorescence technique. Activity of Ca2+ /PL dependent protein kinase C (PKC) was assayed by incubating PKC with histone ⅢS and[γ-32p]ATP. The cytoskeletal proteins were precipitated by Triton and separated by SDS-PAGE. RESULTS: DQD inhibited the platelet aggregation induced by thrombin (500 U/L), when DQD concentrations were 100, 200, and 400 μmol/L, the inhibition rates were 77 % , 86 % , and 82 % respectively. DQD inhibited Ca2+ influx in platelets induced by thrombin (500 U/L) in the presence of extracellular Ca2+ 1 mmol/L in a concentration-dependent manner (10-80 μmol/L); DQD also had inhibitory effect on intracellular Ca2+ mobilization in the absence of extracellular Ca2+ . DQD (10-160 μmol/L) inhibited the cytosolic Ca2+ / PL dependent PKC from platelets in a concentration-dependent manner, but had no effect on membrane PKC. DQD (20 - 200 μmol/L) inhibited the actin polymerization induced by thrombin (500 U/L) in platelets in a concentration-dependent manner. CONCLUSION: DQD inhibited pig platelet aggregation induced by thrombin and its molecular mechanism was due to its inhibition of Ca2+ influx, intracellular Ca2+ mobilization, Ca2+ /目的:研究人工半合成槲皮素—槲皮素-7,4′-二硫酸酯二钠(disodium quercetin-7,4′-disulfate,DQD)对凝血酶(500 U·L^(-1))诱导猪血小板聚集的抑制作用及其机制。方法:用比浊法测定血小板聚集。Fura 2-AM荧光法检测胞浆游离钙浓度([Ca^(2+)]_i)。用组蛋白ⅢS,[γ-^(32)P]ATP与蛋白激酶C(PKC)酶液一起保温的方法测定Ca^(2+)/PL依赖的PKC活性。用SDS-PAGE分离骨架蛋白。结果:DQD对凝血酶诱导的血小板聚集有抑制作用,当DQD的浓度为100,200和400μmol/L时,抑制率分别为77％、86％和82％。DQD(10-80μmol/L)抑制凝血酶诱导的血小板胞外钙内流;DQD对凝血酶诱导的血小板胞内钙动员也有抑制作用。DQD(10-160μmol/L)抑制血小板胞浆Ca^(2+)/PL依赖的PKC,但DQD不影响胞膜PKC。DQD(20-200μmol/L)对凝血酶诱导的血小板肌动蛋白聚合有较强的抑制作用。结论:DQD对凝血酶诱导的猪血小板聚集有抑制作用,其分子作用机制是由于抑制血小板外钙内流、内钙动员、Ca^(2+)/PL依赖的PKC和肌动蛋白聚合。 血小板;;黄酮类;;槲皮素;;血小板聚集;;钙;;蛋白激酶C;;

关 键 词：blood PLATELETS flavones QUERCETIN platelet AGGREGATION calcium protein kinase C ACTINS 

分 类 号：R96[医药卫生—药理学]