Antioxidant mechanism of modified Qiongyu paste against aging based on network pharmacology and experimental validation

摘要

Objective:To provide a possible basis for the anti-aging effect of modified Qiongyu paste(MQP).Methods:Ultra-high-performance liquid chromatography-Q-Orbitrap mass spectrometry was applied to confirm the effective components of MQP that we had identified from databases and research literature.Then,network pharmacology was employed to predict the possible underlying mechanism of MQP against aging.According to the overlap with age-related gene targets,we obtained key targets of MQP against aging using proteineprotein interaction analyses.Superoxide dismutase(SOD)tests were performed in vitro and in vivo.Western blotting of P62 and LC3B and quantitative polymerase chain reaction of the expression of the P62,LC3B,HO-1,Keap1,and Nrf2 genes were conducted on H_(2)O_(2) einduced PC12 cells.Enzyme-linked immunosorbent assays of tumor necrosis factor-a and interleukin-6 were conducted on a D-galactoseeinduced aging mice model.Results:A total of 44 compounds in MQP were finally identified,and 48 gene targets were considered related to anti-aging.Kyoto Encyclopedia of Genes and Genomes and Gene Ontology enrichment analyses linked the principal anti-aging mechanisms of MQP to oxidative resistance,genomic stabilization,and growth hormone regulation.In vitro,H2O2einduced PC12 cells showed that MQP was able to activate SOD,prolong telomeres,and enhance the expression levels of P62,LC3B,HO-1,Keap1,and Nrf2.The quantitative polymerase chain reaction results were affirmed with the reactive oxygen species inhibitor N-acetylcysteine and the reactive oxygen species agonist diallyl-tetrasulfide.In vivo,the D-galeinduced aging mice showed that MQP increased SOD in the brain and reduced tumor necrosis factor-a and interleukin-6 in the serum.Thus,our results suggest that the mechanism of the anti-aging effect of MQP primarily involves antagonizing oxidative stress.Conclusion:From the analyses and experimental validation,we may conclude that oxidative stress resistance may be a potential mechanism of the MQP anti-aging effect.