基于发酵过程中“微生物-成分”关联分析胆南星炮制机制  

Study on processing mechanism of Arisaema cum Bile based on“microorganism-component”correlation analysis of fermentation method

作　　者：武旭 王晶 付毛毛 任润馨 邹婷 张萌萌 张桥 杜升旗 王昌利 孙静 赵重博 WU Xu;WANG Jing;FU Maomao;REN Runxin;ZOU Ting;ZHANG Mengmeng;ZHANG Qiao;DU Shengqi;WANG Changli;SUN Jing;ZHAO Chongbo(College of Pharmacy,Shaanxi University of Chinese Medicine/Shaanxi Provincial Engineering Technology Research Center for Traditional Chinese Medicine Decoction Pieces,Xianyang 712046,China;Traditional Chinese Medicine Processing Techniques Heritage Base(Shaanxi),National Administration of Traditional Chinese Medicine,Xianyang 712046,China;School of Chinese Materia Medica,Tianjin University of Traditional Chinese Medicine/Traditional Chinese Medicine Processing Techniques Heritage Base(Tianjin),National Administration of Traditional Chinese Medicine,Tianjin 301617,China;Shaanxi Xingshengde Pharmaceutical Co.,Ltd.,Tongchuan 727031,China)

机构地区：[1]陕西中医药大学药学院/陕西省中药饮片工程技术研究中心,陕西咸阳712046 [2]国家中医药管理局中药炮制技术传承基地(陕西),陕西咸阳712046 [3]天津中医药大学中药学院/国家中医药管理局中药炮制技术传承基地(天津),天津301617 [4]陕西兴盛德药业有限责任公司,陕西铜川727031

出　　处：《中草药》2025年第3期795-807,共13页Chinese Traditional and Herbal Drugs

基　　金：陕西省高水平中医药重点学科中药炮制学(陕中医药函[2023]83号);陕西省重点研发计划(2024SF-ZDCYL-03-11);秦创原铜川“科学家+工程师”队伍建设项目(2024KJ-KXJGCS-01);王昌利全国老药工传承工作室(国中医药人教函[2024]255号);秦创原中医药产业创新聚集区项目(L2024-QCY-ZYYJJQ-X34)。

摘　　要：目的研究牛胆汁发酵天南星Arisaematis Rhizoma制备胆南星Arisaema cum Bile过程中微生物群落及化学成分的变化规律,为其炮制机制提供研究思路。方法采用新鲜牛胆汁发酵炮制天南星[药物-辅料1∶3,发酵温度(30±5)℃,发酵湿度(70±5)%],制备不同发酵时间的胆南星样品。利用HPLC法对各样品中夏佛塔苷、异夏佛塔苷、黄嘌呤、尿嘧啶等12个成分进行含量测定,同时采用16S rRNA高通量测序技术对其中微生物组成进行鉴定和考察。对“微生物-成分”进行相关性分析,探讨两者在发酵炮制过程中的变化规律。结果随着发酵时间的增加,腺嘌呤、鸟苷、夏佛塔苷、腺苷、肌苷、胸腺嘧啶核苷、甘氨胆酸含量降低;胆酸含量增加;次黄嘌呤、黄嘌呤含量先增加后减少。菌群分析发现,在门水平上主要以变形菌门、厚壁菌门和蓝藻菌门为主;在属水平上主要以乳酸杆菌和假单胞菌为主。关联分析表明,胆南星中化学成分与菌群(门、属)间存在显著相关,其中腺嘌呤、胸腺嘧啶核苷与蓝藻菌门、放线菌门呈显著正相关(P<0.01),与变形菌门呈显著负相关(P<0.05);胆酸与厚壁菌门呈显著正相关(P<0.01),与变形菌门呈显著负相关(P<0.05)。结论牛胆汁发酵炮制天南星的过程中化学成分与微生物群落之间存在显著的变化规律,为其炮制机制研究提供理论基础。Objective To study the changes of microbial community and chemical composition in the preparation of Arisaema cum Bile by fermentation of Arisaematis Rhizoma from bovine bile,and to provide research ideas for its processing mechanism.Methods Firstly,the fresh bovine bile was used to ferment and process Arisaema Rhizoma[drug-excipient ratio=1:3,fermentation temperature(30±5)℃,fermentation humidity(70±5)%]to prepare Arisaema cum Bile samples with different fermentation time.Then,HPLC was used to determine the content of 12 components including schaftoside,isoschaftoside,xanthine and uracil in the samples.As well,the 16S rRNA high-throughput sequencing technology was employed to identify and to investigate the species and the abundance of microbial communities in each Arisaema cum Bile sample.Finally,the correlation analysis of“microorganism-component”was carried out via mathematical algorithm,and the changed rules of them in the processing was discussed.Results With the increase of fermentation time,the contents of adenine,guanosine,schaftoside,adenosine,inosine,thymidine and glycocholic acid decreased,the content of cholic acid increased,the content of hypoxanthine and xanthine increased first and then decreased.The results of flora analysis showed that there were mainly Proteobacteria,Firmicutes and Cyanobacteria at the phylum level.At the genus level,there were mainly Lactobacillus and Pseudomonas.Correlation analysis showed that there was a significant correlation between the chemical component of Arisaema cum Bile and the fermentation flora(phylum and genus).Adenine and thymidine were significantly positively correlated with Cyanophyta and Actinobacteria(P<0.01),while negatively correlated with Proteobacteria(P<0.05).Cholic acid was positively correlated with Firmicutes(P<0.01),and negatively correlated with Proteobacteria(P<0.05).Conclusion There were significant changes between chemical components-microbial communities during the fermentation of Arisaematis Rhizoma with bovine bile processing.The res

关 键 词：胆南星 发酵 微生物群落 化学成分 相关性分析 腺嘌呤 黄嘌呤 尿嘧啶 次黄嘌呤 腺苷 鸟苷 肌苷 胸腺嘧啶核苷 异夏佛塔苷 夏佛塔苷 胆酸 甘氨胆酸 

分 类 号：R283.6[医药卫生—中药学]