Clarification of mechanism of Anti-fatigue Active Components Derived from Earthworms Based on UHPLC-Q-TOF-MS/MS and Network Pharmacology
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Abstract:
The anti-fatigue mechanism of the active constituents of earthworm was clarified using ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry/mass spectrometry (UHPLC-Q-TOF-MS/MS and network pharmacology. UHPLC-Q-TOF-MS/MS was used to identify chemical constituents of earthworm extract. The targets of these constituents and fatigue and anti-fatigue associated genes were collected to identify the common targets. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed. KEGG Mapper was utilized to preliminarily elucidate the anti-fatigue mechanism, and animal experiments were performed to verify the antifatigue efficacy of earthworm extract. Network analysis showed that the anti-fatigue effects of earthworm extract mainly function via positive regulation of phosphorylation, NF-κB signaling pathway, serotonergic synapse signaling, JAK-STAT pathway, and arginine and proline metabolic pathways. As shown in the results of animal experiments, the time of weight-bearing swimming in mice in the middle-dose earthworm extract group was 34.07 min, significantly longer than that (9.78 min) of the CK group (P<0.001). Hepatic glycogen content in the high-dose earthworm extract group was 2 591.21 mg/g, which was significantly higher than that in the CK group (P<0.001). Urea nitrogen and blood lactic acid contents in the high-dose earthworm extract group were 3.65 nmol/L and 182.21 nmol/L, respectively, which was significantly lower than those of the CK group (P<0.01). In conclusion, the antifatigue effect of earthworm extract was exerted through synergistic effects of multiple constituents, targets, and pathways, providing valuable insights and methods for further research into anti-fatigue mechanisms.
