蝉虫草及其寄生真菌产生免疫抑制剂多球壳菌素能力的鉴定  

Identification of ability of Cordyceps and its parasitic fungi to produce immunosuppressant myriocin

作　　者：章安安 贺庆利 ZHANG An'an;HE Qingli(Research Center for Chiral Drugs,Innovative Institute of Chinese Pharmacy,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学创新中药研究院手性药物研究中心,上海201203

出　　处：《上海中医药杂志》2023年第9期11-18,共8页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(22177075)。

摘　　要：目的寻找能产生免疫抑制剂多球壳菌素(myriocin)的蝉虫草或其寄生真菌。方法选取蝉虫草原药材大蝉草和蝉花干燥品,蝉花寄生真菌蝉棒束孢菌(Cordyceps cicadae或Isaria cicadae Miq.BNCC114807)以及一种线虫草属蝉寄生真菌Ophiocordyceps sp.FU30231,通过聚合酶链式反应(PCR)法扩增内源转录间隔区(ITS)序列,鉴定各材料的菌种;分别以乙醇回流提取法和80%甲醇浸提法制备虫生真菌发酵液,并进行液相色谱-质谱(LC-MS)分析;以多球壳菌素标准品为对照,寻找生产多球壳菌素的虫生真菌;通过大量发酵和分离鉴定多球壳菌素,最终确定其产源虫生真菌。结果ITS序列分析表明,蝉虫草原药材大蝉草和蝉花的寄生真菌分别为奇异弯颈霉(Tolypocladium paradoxum)和蝉棒束孢菌。提取液LC-MS分析结果表明,仅Ophiocordycepssp.FU30231中检测到多球壳菌素的产生。大量发酵并分离纯化后,采用核磁谱鉴定确证了Ophiocordycepssp.FU30231产多球壳菌素的能力。结论Ophiocordycepssp.FU30231首次被鉴定为免疫抑制剂多球壳菌素的生产菌株。Objective To identify the source of immunosuppressant myriocin from Cordyceps and its parasitic fungi.Methods Dried raw materials of traditional Chinese medicine,Dachancao and Chanhua,and two parasitic fungi of cicada,Cordyceps cicadae(or Isaria cicadae Miq.BNCC114807)and Ophiocordyceps sp.FU30231,were selected as potential myriocin producing sources.Polymerase chain reaction(PCR)was applied to obtain internal transcribed space(ITS)sequences of strains of all materials.The fermentation broth of entomogenous fungi was prepared by ethanol-reflux extraction and 80%methanol extraction,respectively,and analyzed by LC-MS.The entomogenous fungi producing myriocin were identified by using myriocin as a control standard.After extensive fermentation and isolation of myriocin,the source of myriocin was finally confirmed.Results ITS sequences analysis showed that the parasitic fungi of Dachancao and Chanhua were Tolypocladium paradoxum and Cordyceps cicadae,respectively.The results of LC-MS analysis of the extracts indicated that the production of myriocin was detected only in the extract of Ophiocordyceps sp.FU30231.After massive fermentation,isolation and purification,the ability of Ophiocordyceps sp.FU30231 to produce myriocin was confirmed by NMR spectrum.Conclusion For the first time,Ophiocordyceps sp.FU30231 was identified as the immunosuppressant myriocin-producing fungus which parasitized on cicada.

关 键 词：蝉虫草 虫生真菌 免疫抑制剂 多球壳菌素 免疫疾病 

分 类 号：R282.5[医药卫生—中药学]