Integrating serum pharmacochemistry and network pharmacology to explore the potential mechanisms of Xianshen formula in the prevention of exercise-induced fatigue.

Objective: On the basis of exploring the efficacy of XSF, the material basis and mechanism of XSF for the prevention of exercise-induced fatigue were clarified by network pharmacology using blood-absorbed components as the research object.

Methods: UPLC-Q-TOF/MS was used to identify the blood components of XSF. On this basis, the target prediction of the blood-entering components was obtained from the Swiss Target Prediction and SuperPred database, and the target related to exercise-induced fatigue was acquired from OMIM, GeneCards, and other disease databases. The network model of " components- targets-diseases" of XSF was established by Cytoscape software. The String data analysis platform was used to build the PPI network to filter out the primary targets. The DAVID database was used for gene ontology (GO) enrichment analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the key targets.

Results: Administration of XSF increased swimming time, decreased BLA, BUN, and MDA levels, and elevated SOD levels in swimming exhausted rats and mice. A total of 60 in vitro active ingredients, 45 blood-entry prototype ingredients, and 41 blood-entry metabolized ingredients were identified. Out of 717 possible targets of action between medication components and exercise exhaustion, network pharmacology identified 176 nodes with a maximum value of TP53 (Degree = 81) according to PPI analysis. The key targets were involved in 96 KEGG pathways including the PI3K-Akt signaling pathway, MAPK signaling pathway, HIF-1 signaling pathway TNF signaling pathway, and 170 GO pathways. The top 10 targets in the "component-target-pathway" network of XSF against EIF were predicted to be NFKB1, PIK3R1, GRIN1, CACNA1B, SLC6A5, NTRK3, GRIA2, TRIM24, TOP2A, TLR4 and RORB, TLR4 and RORB.

Conclusion: XSF is effective in the prevention of EIF and its potential pharmacological mechanisms may be related to the improvement of energy metabolism, regulation of inflammatory response, and regulation of oxidative stress.