The objective of this study is to investigate the regulation effects of the active ingredients in Gegenqinlian Decoction (GD) on the intestinal mucosal flora of mice with diarrhea induced by high temperature and humidity based on systems pharmacology approach. Fifteen mice were randomly assigned to three equal groups of five mice, namely control (ctcm) group, model (ctmm) group and treatment (cttm) group. Mice in the cttm group were given 20 mL/kg of GD and sterile water was used as a placebo control twice a day for four consecutive days. We used the third-generation molecular high-throughput sequencing technology to measure the intestinal mucosal flora changes in mice. Combined with network pharmacology to predict the medicinal substances and action targets of GD against diarrhea. Results showed that Operational Taxonomic Unit (OTU) number and Alpha diversity in the intestinal mucosal flora of cttm group recovered and higher than that of the ctcm group. There were differences in the community structure between the ctmm and cttm groups in the Principal Coordinates Analysis (PCoA). The relative abundance results indicated dominant bacteria species (such as Lactobacillus crispatus, Muribaculum intestinal, Neisseria mucosa) in the intestinal mucosa of the three groups. Moreover, we screened out 146 active ingredients in GD corresponding to 252 component targets, and 328 disease targets in diarrhea to obtain 31 drug–disease common targets. Protein–protein interaction (PPI) networks mainly involved the core proteins such as Tumor necrosis factor (TNF) and Interleukin-6 (IL-6). Enrichment analyses showed that GD played a role in the treatment of diarrhea by regulating the hypoxia inducible factor-1 (HIF-1), vascular endothelial growth factor (VEGF) and adipocytokine signaling pathways and so on. In brief, the active ingredients of GD could intervene from oxidative stress and inflammatory response through multiple targets and multiple channels to adjust the balance of intestinal mucosa flora, thereby playing a role in the treatment of diarrhea.

本研究旨在基于系统药理学方法探讨葛根芩连汤（GD）有效成分对高温高湿腹泻小鼠肠黏膜菌群的调节作用。将15只小鼠随机分为三组，每组5只小鼠，即对照组(ctcm)、模型组(ctmm)和治疗组(cttm)。cttm组小鼠给予20mL/kg的GD，并使用无菌水作为安慰剂对照，每天两次，连续四天。我们采用第三代分子高通量测序技术来测量小鼠肠粘膜菌群的变化。结合网络药理学预测GD抗腹泻的药用物质及作用靶点。结果显示，cttm组肠粘膜菌群的操作分类单元（OTU）数和Alpha多样性有所恢复，且高于ctcm组。主坐标分析（PCoA）中ctmm和cttm组之间的群落结构存在差异。相对丰度结果显示三组肠粘膜中均存在优势菌种（如卷曲乳杆菌、肠粘膜杆菌、粘膜奈瑟菌）。此外，我们筛选出GD中146个活性成分对应252个成分靶点，以及腹泻中的328个疾病靶点，获得31个药物-疾病共同靶点。蛋白质-蛋白质相互作用（PPI）网络主要涉及肿瘤坏死因子（TNF）和白细胞介素-6（IL-6）等核心蛋白质。富集分析表明，GD通过调节缺氧诱导因子1（HIF-1）、血管内皮生长因子（VEGF）和脂肪细胞因子信号通路等发挥治疗腹泻的作用。总之，GD的活性成分可以通过多靶点、多途径干预氧化应激和炎症反应，调节肠粘膜菌群平衡，从而发挥治疗腹泻的作用。