广地龙分子鉴定及与赤子爱胜蚓在不同提取方法下蚓激酶效价比较  

Molecular identification of Pheretima and comparison of its lumbrokinase potency with that of Eisenia foetida under different extraction methods

作　　者：赵梓颐 武佳文 白秋林 胡琦敏 黄云峰 ZHAO Ziyi;WU Jiawen;BAI Qiulin;HU Qimin;HUANG Yunfeng(Guangxi Institute of Chinese Medicine and Pharmaceutical Science/Guangxi Key Laboratory of Traditional Chinese Medicine Quality Standards,Nanning 530022,China;College of Horticulture and Landscape Architecture,Northeast Agricultural University,Harbin 150000,China;Youjiang Medical University for Nationalities,Baise 533000,China)

机构地区：[1]广西壮族自治区中医药研究院/广西中药质量标准研究重点实验室,南宁530022 [2]东北农业大学园艺园林学院,哈尔滨150000 [3]右江民族医学院,广西百色533000

出　　处：《吉林中医药》2025年第1期104-108,共5页Jilin Journal of Chinese Medicine

基　　金：广西中医药重点学科建设项目(GZXK-Z-20-69);广西中医药适宜技术开发与推广项目(GZSY2024008);广西壮族自治区中医药管理局自筹经费科研课题(GXZYA20220014)。

摘　　要：目的探讨COI条形码鉴定和区分广地龙基原参环毛蚓和赤子爱胜蚓的适用性,探讨提取方法和物种对蚓激酶效价的影响。方法提取参环毛蚓和赤子爱胜蚓DNA,PCR扩增COI片段后进行测序和NCBI数据库比对。利用匀浆法和电击法获取参环毛蚓和赤子爱胜蚓蚓激酶粗提液。冷冻干燥后用生理盐水分别稀释成相同浓度,在纤维蛋白平板上测定和比较蚓激酶效价。结果COI条形码技术能够区分参环毛蚓和赤子爱胜蚓。提取方式相同,参环毛蚓蚓激酶效价高于赤子爱胜蚓。物种相同,电击法获得的蚓激酶效价均更高。结论序列比对结果与已知物种信息一致,使用电击法获取参环毛蚓蚓激酶具有更高效价。Objective To explore the applicability of COI barcodes in identifying the original worm of Pheretima aspergillurn and distinguishing it from Eisenia fetida,and to investigate the effects of extraction methods and species on the potency of earthworm kinase.Methods The DNAs of Pheretima aspergillurn and Eisenia fetida were extracted,and the COI fragments were amplified by PCR,then sequenced for a comparison with the information in the NCBI database.The crude extracts of earthworm kinase from Pheretima aspergillurn and Eisenia fetida were obtained using the homogenization method and electric shock method.After freeze-drying,the extracts were diluted with normal saline to the same concentration,and then the potency of earthworm kinase was measured and compared on fibrin plates.Results The COI barcode technique distinguished between Pheretima aspergillurn and Eisenia fetida.Employing the same extraction method,the kinase potency from Pheretima aspergillurn was always higher than that of Eisenia foetida.The efficacy of earthworm kinase obtained by electric shock was higher for the same species.Conclusion The sequence alignment results are consistent with the information of known species,and the use of electric shock method ensures to obtain earthworm kinase with higher potency.

关 键 词：参环毛蚓 赤子爱胜蚓 分子鉴定 蚓激酶 效价 

分 类 号：R282.5[医药卫生—中药学]