基于高通量测序和化学轮廓分析研究黄连在连栀矾溶液发酵炮制中的作用  

Based on high-throughput sequencing and chemical profiling to study role of Coptidis Rhizoma in concocting of Lianzhifan solution

作　　者：吴泽 杜娟[2] 谢洁 勾洵[1] 彭玲 叶羊 张晓喻[1] 袁学刚 袁钰婷 史迎欣 WU Ze;DU Juan;XIE-Jie;GOU Xun;PENG Ling;YE Yang;ZHANG Xiao-yu;YUAN Xue-gang;YUAN Yu-ting;SHI Ying-xin(College of Life Science,Sichuan Normal University,Chengdu 610101,China;College of Geography and Resource Science,Sichuan Normal University,Chengdu 610101,China;The Sixth People’s Hospital of Chengdu,Chengdu 610058,China)

机构地区：[1]四川师范大学生命科学学院,四川成都610101 [2]四川师范大学地理与资源科学学院,四川成都610101 [3]成都市第六人民医院,四川成都610058

出　　处：《中草药》2021年第6期1623-1633,共11页Chinese Traditional and Herbal Drugs

基　　金：四川省科技计划项目(2020YJ0372)。

摘　　要：目的探究黄连Coptidis Rhizoma在连栀矾溶液传统发酵炮制中的作用。方法按照连栀矾溶液传统配方比例,将其拆分为3组配方溶液:连栀矾组(黄连+栀子+白矾)、黄连组(黄连+白矾)和栀子组(栀子+白矾),采用HPLC分析比较3组溶液发酵前后的化学成分变化轮廓,采用Illumina Hiseq高通量测序技术测定并比较发酵溶液中微生物的群落特征。结果传统配方连栀矾经发酵后环烯醚萜类成分发生了较显著的变化(P<0.05),生物碱类成分未发生明显变化,拆分后的黄连组中生物碱类成分和栀子组中环烯醚萜类成分均没有发生明显变化。分别对各组发酵溶液进行测序,根据α多样性分析结果,3组发酵液中真菌多样性为连栀矾组<栀子组<黄连组,真菌的丰富度为栀子组<连栀矾组<黄连组;细菌多样性为连栀矾组<黄连组<栀子组,细菌的丰富度为连栀矾组<栀子组<黄连组。根据β多样性分析结果,3组溶液中的真菌和细菌的群落结构均有明显差异。根据微生物相对丰度分析可知,3组的优势真菌均为子囊菌门、担子菌门和罗兹菌门,不同组间丰度无明显差异;真菌优势属中的孢霉属的丰度在连栀矾组最高,显著高于另外2组(P<0.05),栀子组又显著高于黄连组(P<0.05);韦斯特壳属的丰度在连栀矾组中最低,显著低于黄连组和栀子组(P<0.05),后两者间丰度无明显差异。3组的优势细菌均为变形菌门、厚壁菌门、放线菌门和拟杆菌门,其中连栀矾组中变形菌门的丰度极显著低于黄连组和栀子组(P<0.01),厚壁菌门的丰度在连栀矾组中极显著高于黄连组和栀子组(P<0.01),放线菌门和拟杆菌门的丰度在3组发酵溶液的差异较小;细菌优势菌属中的乳杆菌属在连栀矾组中显著高于黄连组和栀子组(P<0.05),在黄连组中也显著高于栀子组(P<0.05),伯克氏菌属丰度在黄连组和栀子组中无显著差异,都显著高于连栀矾组中的丰度(P<0.05)。�Objective To investigate the role of Huanglian(Coptidis Rhizoma)in the traditional fermentation processing of Lianzhifan solution.Methods According to the traditional formula ratio of Lianzhifan solution,it was divided into three groups of formula solutions:Lianzhifan group[Huanglian(Coptidis Rhizoma)+Zhizi(Gardeniae Fructus)+Baifan(Alumen)(LZF)],Coptidis Rhizoma group[Coptidis Rhizoma+Alumen(CR)],and Gardeniae Fructus group[Gardeniae Fructus+Alumen(GF)].The chemical composition changes of the three groups of solutions before and after fermentation were analyzed and compared by HPLC.Illumina Hiseq high-throughput sequencing technology was used to determine and compare the community characteristics of microorganisms in the fermentation solution.Results The iridoid composition of the traditional formula LZF group has undergone significant changes after fermentation(P<0.05),and the alkaloid composition has not changed significantly.The alkaloid composition in CR and the iridoids in GF did not change significantly after fermentation.The fermentation solutions of each group were sequenced.According to the results ofαdiversity analysis,the fungal diversity in the three fermentation broths was LZF<GF<CR,the fungal richness was GF<LZF<CR;The bacterial diversity was LZF<CR<GF,the abundance of bacteria was LZF<GF<CR.According to the results ofβdiversity analysis,the community structures of fungi and bacteria in the three groups of solutions were significantly different.According to the analysis of the relative abundance of microorganisms,the dominant fungi in the three groups were Ascomycota,Basidiomycota,and Rozellomycota,and there was no significant difference in abundance among different groups.The abundance of Chrysosporium in the dominant genus of fungi was the highest in the LZF group,which was significantly higher than the other two groups(P<0.05),and the GF group was significantly higher than the CR group(P<0.05);The abundance of Westerdykella was the lowest in LZF,significantly lower than CR and GF(P<0.05)groups

关 键 词：连栀矾溶液 黄连 高通量测序 化学轮廓分析 微生物多样性 发酵 炮制 群落特征 环烯醚萜 生物碱 α多样性分析 β多样性分析 真菌 细菌 

分 类 号：R283.1[医药卫生—中药学]