Integrating spectrum-effect relationship, network pharmacology and multi-omics to decipher the mechanism of Cybister chinensis Motschulsky against chronic kidney disease.
ETHNOPHARMACOLOGICAL RELEVANCE Cybister chinensis Motschulsky is a traditional Chinese medicinal insect traditionally used to tonify the kidney, and clinically applied for the management of chronic kidney disease (CKD). AIM OF THE STUDY The present work was dedicated to identifying bioactive ingredients in the ethyl acetate fraction of C. chinensis and elucidating their mechanisms underlying the treatment of CKD. METHODS Compounds were isolated and used to establish fingerprints. Anti-inflammatory and anti-injury activities in vitro and grey relational analysis were applied to establish spectrum-effect relationships. Network pharmacology combined with molecular docking was employed to identify the candidate bioactive components. The effects of ethyl acetate fraction of C. chinensis were explored in doxorubicin-induced CKD rat model via Western blot, RT-PCR, transcriptomics, and metabolomics. Molecular dynamics simulation was applied to assess binding affinity of bioactive components. Cell experiments verified effects of components on inflammation, oxidative stress, and fibrosis. RESULTS The ethyl acetate fraction of C. chinensis was confirmed as the active fraction. Compounds 6, 10, 22, 23 and 27 were screened as the potential active components. Ethyl acetate fraction improved the condition of CKD rats, while multi-omics identified a triple-functional network against inflammation, oxidative stress, and fibrosis, with PI3K-AKT/MAPK/HIF-1 pathways verified. Cell experiments and molecular dynamics simulation identified compound 6 as the key active component, with its regulatory effects validated through PI3K-AKT/MAPK/HIF-1 pathways. CONCLUSION Ethyl acetate fraction of C. chinensis and its active component exerted renoprotective effects by suppressing inflammation, oxidative stress, and fibrosis in CKD via regulating PI3K-Akt, MAPK, and HIF-1 pathways.