Abstract: 【Objective】 The aim of this study was to predict the potential target and mechanism of Mahuang Gancao decoction in the treatment of pulmonary edema,and to verify it through animal experiments in vivo,so as to provide a reference for further research on the pharmacological effects of Mahuang Gancao decoction.【Method】 The potential active ingredients and related targets of Mahuang Gancao decoction were screened by Traditional Chinese Medicine Systems Pharmacology Database (TCMSP).The related targets of pulmonary edema were obtained from the GeneCards database.And then,the intersection of the active ingredient targets and disease targets was selected.The network diagram of ingredient-target was established with Cytoscape 3.6.GO fuction and KEGG pathway enrichment analysis were carried out in DAVID database.The active ingredients with the highest values and the key core targets were docked and verified by AutoDdock Vina to support the prediction results of network pharmacology.Finally,BALB/c mice were used for experimental verification.2% ammonium chloride was used to construct the pulmonary edema model.Mice were randomly divided into blank group,ammonium chloride model group and Mahuang Gancao decoction treatment group.The blank group was intraperitoneally injected with 200 μL normal saline every morning,and the ammonium chloride model group and the Mahuang Gancao decoction treatment group were intraperitoneally injected with 200 μL 2% ammonium chloride solution,the ammonium chloride model group and the blank group were intragastrically administered with 200 μL normal saline every night,and the Mahuang Gancao decoction treatment group was intragastrically administered with 200 μL 1 g/mL Mahuang Gancao decoction.After 5 days of continuous treatment,the mRNA expression levels of interleukin-6 (IL6),inducible nitric oxide synthase (NOS2),mitogen-activated protein kinase 3 (MAPK3) and matrix metalloproteinase 9 (MMP9) genes were detected by Real-time quantitative PCR.【Result】 There were 23 and 92 potential pharmaceutical active ingredients in ephedra and licorice,respectively,and 239 targets of the two components.5 196 targets of pulmonary edema were identified.There were 213 intersecting targets of drugs and diseases.The core targets were RAC-α serine/threonine protein kinase (AKT1),cell tumor antigen p53 (TP53), IL6,vascular endothelial growth factor A (VEGFA),estrogen receptor 1 (ESR1),epidermal growth factor receptor (EGFR),MAPK3,peroxidase synthase 2 (PTGS2),signal transducer and activator of transcription 3 (STAT3),MMP9,NOS2,etc.954 items were obtained by GO function enrichment analysis,including 710 items of biological process,involving metabolic process,gene transcription process,biological regulation process,drug stimulation process,etc.Cell component 92 items,mainly included cell membrane,cytoplasm,nucleus,organelles,etc.152 items of molecular function mainly included enzyme catalysis,protein binding,transcriptional regulation,etc.The results of the KEGG pathway enrichment analysis were involved in 170 signaling pathways,mainly related to tumor necrosis factor (TNF),PI3K-Akt,IL17 and other signaling pathways.Results of molecular docking showed that the key active ingredients had a good binding ability with the core targets.Real-time PCR results showed that compared with the blank group,the mRNA expression of IL6, MAPK3 and MMP9 genes in ammonium chloride model group were significantly or extremely significantly up-regulated (P＜0.05 or P<0.01),and the mRNA expression of NOS2 gene was extremely significantly down-regulated (P＜0.01).Compared with the ammonium chloride model group,the mRNA expression of IL6, MAPK3 and MMP9 genes in Mahuang Gancao decoction treatment group were significantly or extremely significantly down-regulated (P＜0.05 or P<0.01),and the mRNA expression of NOS2 gene was extremely significantly up-regulated (P＜0.01).【Conclusion】 Mahuang Gancao decoction could treat pulmonary edema through multiple components,multiple targets and multiple pathways.Quercetin,kaempferol,naringenin,luteolin and so on,which were active components,might exert inflammatory control and immunomodulatory effects on TNF,PI3K-Akt,IL17 signaling pathways through acting on IL6,MAPK3,MMP9,NOS2 and other targets.

Key words: Mahuang Gancao decoction; pulmonary edema; network pharmacology; molecular docking