Protective effect and mechanism of vaccarin on acute lung injury in sepsis mice
Objective Based on network pharmacology and animal experiments to explore the protective mechanism of vaccarin(VAC)in acute lung injury of sepsis mice.Methods The potential targets of VAC and the disease targets related to pulmonary sepsis were collected through the Swiss Target Prediction database and the GeneCards database separately.Then,the common targets for VAC(1,5 mg·kg-1)and diseases were obtained using Draw Venn Diagram software.The STRING 11.5 database protein-protein interaction was used to construct PPI network and a drug target disease network was constructed using Cytoscape 3.10.0.Finally,Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis for key targets were conducted utilizing the Metascape database.A sepsis model was established by intraperitoneal injection of lipopolysaccharide(LPS)and treated with VAC or dexamethasone(DEX).Then,lung tissue and serum were collected from each group,and HE,Masson,and TUNEL staining were used to observe changes in lung tissue morphology,fibrosis area,and cell apoptosis.The levels of inflammatory factors in serum and lung tissue were detected by ELISA and qRT-PCR,respectively.The expression of inflammatory pathway related proteins in lung tissue was detected by immunohistochemistry and Western blotting.Results There were 44 common targets for VAC and pulmonary sepsis.A total of 924 GO items were obtained by GO enrichment analysis,including 823 for biological processes,52 for cellular composition,and 49 for molecular functions.KEGG analysis identified 20 signaling pathways,including cancer pathway,PI3K-Akt,and JAK-STAT signaling pathway.The animal experiment results showed that compared with the control group,the model group mice had severe lung tissue damage and fibrosis,lung index significantly increased(P＜0.05),and the expression of inflammatory factor in serum and tissue was increased(P＜0.01,0.001).While,compared with model group,VAC and DEX improved pathological morphology of lung tissue,reduced fibrosis and the expression levels of serum TNF-α,IL-1β,IL-6(P＜0.05,0.01,0.001).The expression of inflammatory proteins in lung tissue,and the number of cell apoptosis decreased,while the expression of P13K and AKT1 proteins increased(P＜0.05,0.01,0.001).The results of animal experiments were consistent with the results of network pharmacology.Conclusion VAC have a certain protective effect on acute lung injury in sepsis mice,and its mechanism may be related to the regulation of PI3K-Akt and NLRP3/TNF-αpathways related to its occurrence and development,which provides a basis for the in-depth study of the anti-inflammation mechanism of VAC.
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