尖吻蝮蛇蛇毒降纤酶的分离纯化及其经直肠给药的实验研究  

Isolation and purification of agkistrodon venom defibrase and experimental study of its transrectal dosing

作　　者：孔露平 廖明 张学荣 周怡 张昊 胡少聪 肖曼琪 张子彦 罗小玲 KONG Luping;LIAO Ming;ZHANG Xuerong;Zhou Yi;ZHANG Hao;HU Shaocong;XIAO Manqi;ZHANG Ziyan;LUO Xiaoling(School of Basic Medical Sciences Guangxi Medical University,Guangxi Nanning 530021,China;Institute of Life Sciences Guangxi Medical University,Guangxi Nanning 530021,China;Department of Experimental Research,Affiliated Cancer Hospital,Guangxi Medical University,Guangxi Nanning 530021,China)

机构地区：[1]广西医科大学,基础医学院,广西南宁530021 [2]广西医科大学,生命科学研究院,广西南宁530021 [3]广西医科大学附属肿瘤医院实验研究部,广西南宁530021

出　　处：《中国医院药学杂志》2024年第9期1016-1022,共7页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金资助项目(编号:82260386)。

摘　　要：目的:通过层析法从尖吻蝮蛇蛇毒中分离纯化降纤酶,探索降纤酶经直肠给药进入血液循环而降低体内纤维蛋白原水平的可行性。方法:从尖吻蝮蛇蛇毒粗毒中分离降纤酶,经小鼠直肠给药后动物成像观察降纤酶在小鼠脏器中的分布,病理学检测降纤酶对组织的影响,质谱测定小鼠血浆成分,ELISA检测血浆纤维蛋白原水平。结果:尖吻蝮蛇蛇毒粗毒分离纯化获得电泳纯的降纤酶,经直肠给药后其安全性高,无急性毒性反应,直肠组织无明显病理改变。免疫组化结果显示降纤酶分布于直肠组织中(P<0.01),Shotgun质谱测得给药1 h后小鼠血浆存在降纤酶片段,动物成像显示给药3 h后染料标记的降纤酶分子分布大量分布于盲肠组织中,并随时间的延长其含量逐步减少。降纤酶经直肠给药能降低体内纤维蛋白原水平(P<0.01)。结论:尖吻蝮蛇蛇毒粗毒经二步层析法分离获得电泳纯的降纤酶;降纤酶可经直肠给药后吸收进入血液循环并降低体内纤维蛋白原水平。OBJECTIVE To isolate and purify defibrase from agkistrodon venom by chromatography and explore the feasibility of transrectal dosing into blood circulation to reduce the level of fibrinogen in vivo.METHODS Defibrase was isolated from crude venom of Agus agicoides.After a transrectal injection,the distribution of defibrase in the organs of mice was observed by animal imaging.The effect of defibrase on tissues was detected by pathology examination.RESULTS Electrophoretically pure defibrillinase was acquired by isolation and purification of crude venom of sharp-nosed pit viper.It was safe after transrectal dosing without acute toxicity or obvious pathological changes in rectal tissues.Immunohistochemistry indicated that fibrinogenin was distributed in rectal tissues(P<0.01).Shotgun mass spectrometry was utilized for measuring the plasma level of fibrinogen fragments after 1 h of dosing.Animal imaging revealed that dye-labeled fibrinogen was predominantly distributed in cecal tissues after 3 h of dosing with a gradual decrease.Rectal administration of fibrinogen reduced fibrinogen level in vivo(P<0.01).CONCLUSION Electrophoretic pure defibrase is acquired from crude venom of agkistrodon by two-step chromatography.Defibrase may be absorbed into blood circulation and reduce fibrinogen level after transrectal dosing.

关 键 词：降纤酶 尖吻蝮蛇蛇毒 直肠给药 分离纯化 

分 类 号：R991[医药卫生—毒理学]