眼镜蛇毒金属蛋白酶atrase A抗血小板聚集作用及机制  

Inhibitory effect and mechanism of cobra venom metalloproteinase atrase A on platelet aggregation

作　　者：王彩娥[1,2] 孙黔云[2] 李敏[3] 石京山[1] 

机构地区：[1]遵义医学院,贵州遵义563000 [2]贵州省中国科学院天然产物化学重点实验室,贵州贵阳550002 [3]贵州省人民医院,贵州贵阳550002

出　　处：《中国药理学通报》2009年第6期743-747,共5页Chinese Pharmacological Bulletin

基　　金：国家重点基础研究发展计划(973计划)资助项目(No2007CB516813,2009CB526512);国家自然科学基金资助项目(No30560035);贵州省科技攻关资助项目(No黔科合工计字2002-1075号);贵州省优秀青年科技人才资助项目(No黔科合人字2005-0510号);贵州省优秀科技教育人才省长专项资金资助项目;贵州省中药现代化科技产业研究开发专项资助项目(No2006-5041号)

摘　　要：目的研究中华眼镜蛇毒金属蛋白酶atrase A对血小板聚集的影响及其相关的机制。方法测定atrase A对二磷酸腺苷、胶原、血小板活化因子、花生四烯酸、瑞斯托霉素、凝血酶诱导血小板聚集的影响情况,并通过蛋白质免疫印迹检测atrase A对血小板膜糖蛋白和血管假血友病因子的酶切情况。结果中华眼镜蛇毒金属蛋白酶atrase A能明显抑制由瑞斯托霉素和凝血酶诱导的血小板聚集,这种抑制作用呈量效、时效关系。而atrase A和血小板预孵5min后对二磷酸腺苷、胶原、血小板活化因子、花生四烯酸诱导的血小板聚集有微弱的抑制作用,预孵时间延长至30min对血小板聚集有明显的抑制作用。蛋白质免疫印迹结果显示atrase A能特异性酶切血小板膜糖蛋白GPIb,但对vWF几乎没有酶切作用。结论中华眼镜蛇毒金属蛋白酶atrase A对二磷酸腺苷、胶原、血小板活化因子、花生四烯酸、瑞斯托霉素、凝血酶诱导的血小板聚集均有抑制作用,其中对瑞斯托霉素和凝血酶诱导的血小板聚集具有明显的抑制作用,其机制是通过酶切血小板膜糖蛋白GPIb。Aim To investigate the effect and mechanism of cobra venom metalloproteinase atrase A on inhibiting platelet aggregation. Methods Platelet aggregation induced by collagen, ADP, PAF, AA, ristoeetin and thrombin,respectively,was measured turbimetrieally after platelet incubated with atrase A. Western blot was used to detect the effect of atrase A on cleavage of platelet membrane glycoprotein and yon Willebrand Factor. Results Atrase A significantly inhibited platelet aggregation induced by ristocetin and thrombin in a dose-and time-dependent manner. Meanwhile,atrase A just showed slight inhibitory effect on platelet aggregation induced by PAF, AA, collagen, and ADP after incubated with PRP for 5 min. After incubation time was prolonged to 30 min, significant inhibition was shown on platelet aggregation. Western blot revealed that atrase A cleaved platelet membrane glycoprotein GPIb. Conclusions Cobra venom metalloproteinase atrase A significantly inhibits platelet aggregation induced by ristocetin and thrombin due to the cleavage of platelet membrane glycoprotein Ib. And atrase A also has inhibitory effect on platelet aggregation induced by ADP, PAF, AA, and collagen.

关 键 词：中华眼镜蛇毒 蛇毒金属蛋白酶 血小板聚集 血小板膜糖蛋白 瑞斯托霉素 凝血酶 

分 类 号：R331.124[医药卫生—人体生理学] R973.2[医药卫生—基础医学]