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）。结论 连栀矾溶液配方中的黄连能提高其中栀子的有效成分的发酵转化，并且是通过对发酵体系中的优势菌属的丰度进行调节而发挥作用的。进一步佐证了传统配方制剂的合理性和科学性。

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 of α diversity 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 of β diversity 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, and there was no significant difference in abundance between the latter two. The dominant bacteria in the three groups were Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes. The abundance of Proteobacteria in LZF was significantly lower than that of CR and GF (P < 0.01). The abundance of Firmicutes in LZF was extremely significantly higher than that of CR and GF (P < 0.01). The abundance of Actinobacteria and Bacteroidetes had smaller differences in the three fermentation solutions. Lactobacillus in the dominant bacterial genus was significantly higher in LZF than CR and GF (P < 0.05), and that in CR was also significantly higher than GF (P < 0.05), there was no significant difference in Burkholderia abundance in CR and GF, both were significantly higher than the abundance in LZF (P < 0.05). Conclusion Coptidis Rhizoma in the LZF formula can improve the fermentation conversion of the active ingredients of Gardeniae Fructus, and it works by adjusting the abundance of dominant bacteria in the fermentation system. This also further supports the rationality and scientificity of traditional formulations.

R283.1

四川省科技计划项目（2020YJ0372）