Studies Combining Network Pharmacology with In Vivo Experiments Uncover the Fever-reducing Effects and Underlying Molecular Pathways of Radix Isatidis.

Abstract

Objective: The objective of this investigation was to examine the mechanism through which Radix isatidis operates, utilizing network pharmacology and molecular docking techniques.

Methods: Active components and associated targets of Radix isatidis were identified using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) alongside the GeneCards database. A protein-protein interaction (PPI) network connecting the targets of the active ingredients with those related to febrile diseases was constructed through STRING. The analysis of core nodes was carried out using Cytoscape software, followed by a further exploration of the PPI network using the DAVID database. Lastly, the underlying mechanism of the antipyretic action was also examined utilizing the DAVID database. Functional annotation through Gene Ontology (GO) and enrichment analysis of pathways from the Kyoto Encyclopedia of Genes and Genomes (KEGG) were conducted utilizing the DAVID database, with essential components chosen for molecular docking through Pymol software. Mice were injected intraperitoneally with lipopolysaccharide (LPS) and treated by continuous gavage with Radix isatidis. They were then evaluated using temperature monitoring, blood tests, organ index calculation, PI3K-AKT pathway protein assays, and inflammatory factor reverse transcription polymerase chain reaction (RT-PCR) assays.

Results: Twelve active components of Radix isatidis were screened, and 107 genes were identified at the intersection of Radix isatidis and fever. These genes were found to be involved in the PI3K-AKT signaling pathway, proteoglycans in cancer, and mechanisms related to blood lipids and atherosclerosis. The top nine targets screened by constructing a PPI network were IL6, AKT1, EGFR, STAT3, CASP3, ESR1, PTGS2, PPARG, and MAPK3, which indicated that Radix isatidis may play a protective role by affecting the PI3K/AKT-related signaling pathway. In in vivo experiments, Radix isatidis inhibited the activation of PI3K/AKT-related pathways, reduced inflammatory responses, and improved symptoms in mice treated with LPS. Conclusion The molecular mechanism of action of the antipyretic effect of Radix isatidis was predicted through network pharmacology, and it was also verified in vivo, as Radix isatidis reduced the inflammatory response, lowered the body temperature of mice, and protected LPStreated mice via effects on the PI3K-AKT-related signaling pathway.