两种蝎毒多肽的分离纯化及抗血栓活性  

Purification and antithrombotic activity of two scorpion venom peptides

作　　者：孙淑仃 王清[3] 张殿文[3] 张文婷 付宏征[2] 袁丹[1] 李伟[3] SUN Shuding;WANG Qing;ZHANG Dianwen;ZHANG Wenting;FU Hongzheng;YUAN Dan;LI Wei(College of Traditional Chinese Medicine,Shenyang Pharmaceutical University,Shenyang 110016,China;State Key Laboratory of Natural and Biomimetic Drugs,Peking University Health Science Center,Beijing 100191,China;Jilin Academy of Traditional Chinese Medicine,Jilin 130021,China;Beijing Key Laboratory of Quality Assessment Of Traditional Chinese Medicine,School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100102,China)

机构地区：[1]沈阳药科大学中药学院,辽宁沈阳110016 [2]北京大学医学部天然药物及仿生药物国家重点实验室,北京100191 [3]吉林省中医药科学院,吉林吉林130021 [4]北京中医药大学中药学院中药品质评价北京市重点实验室,北京100102

出　　处：《沈阳药科大学学报》2021年第1期45-53,共9页Journal of Shenyang Pharmaceutical University

摘　　要：目的筛选出蝎毒的抗血栓成分。方法采用反相色谱技术对蝎毒进行分离纯化,通过质谱分析及数据库检索对得到的流分进行鉴定。选取小鼠黑尾试验与大鼠体内血栓形成试验,以及斑马鱼血栓模型试验对得到的单体多肽进行抗血栓活性筛选。结果蝎毒经色谱分离纯化得34 min和48 min两个流分,分别记为34号样和48号样。经质谱分析34号样为BmK-M10,分子质量为7039.25 Da;48号样为BmKITc,分子质量为6909.89 Da。对BmK-M10和BmKITc进行抗血栓活性筛选,小鼠黑尾试验与大鼠体内血栓形成试验表明,两者均有抗血栓活性,以BmKITc效果最佳。斑马鱼抗血栓活性筛选试验表明,BmK-M10和BmKITc均有抗血栓活性,其中BmK-M10随着浓度增高,尾部血管红细胞染色强度值增大,呈现反向结果;BmKITc随着浓度增高,尾部血管红细胞染色强度值减小,呈浓度依赖性结果。结论蝎毒多肽BmK-M10和BmKITc均具有抗血栓活性,这为蝎毒治疗抗血栓类疾病提供了依据。Objective To extract and screen out the antithrombotic components of scorpion venom.Methods The scorpion venom was separated and purified by reversed-phase chromatography,and the obtained fractions were identified by mass spectrometry and database search.The black tail experiment in mice,the thrombosis experiment in rats,and the zebrafish thrombosis model experiment were used to screen the obtained monomeric peptides for antithrombotic activity.Results The components of scorpion venom were separated and purified by chromatography for 34 min and 48 min single fractions,and the two single fractions were recorded as No.34 sample and No.48 sample,respectively.After mass spectrometry,the No.34 sample was identified as BmK-M10 with a molecular weight of 7039.25 Da,and the No.48 sample was identified as BmKITc with a molecular weight of 6909.89 Da.BmK-M10 and BmKITc were screened for antithrombotic activity.The black tail experiment in mice and the thrombosis experiment in rats showed that both the two substances had antithrombotic activity,and BmKITc had the better effect.The zebrafish antithrombotic activity screening experiment showed that both BmK-M10 and BmKITc had antithrombotic activity.As the concentration of BmK-M10 increased,the red blood cell staining intensity of the tail blood vessels increased,showing the reverse result;as the concentration increased,the staining intensity of vascular red blood cell in the tail vessels decreased,showing a concentration-dependent result.Conclusion BmK-M10 and BmKITc have antithrombotic activity,which will provide the basis for scorpion venom antithrombotic disease treatment.

关 键 词：蝎毒 抗血栓 BmK-M10 BmKITc 尾部血管红细胞染色强度 

分 类 号：R917[医药卫生—药物分析学] R96[医药卫生—药学]