Integrated UPLC-ESI-MS/MS, network pharmacology, and transcriptomics to reveal the material basis and mechanism of Schisandra chinensis Fruit Mixture against diabetic nephropathy.

Backgrounds: It has been regarded as an essential treatment option for diabetic nephropathy (DN) in Traditional Chinese medicine. Previous studies have demonstrated the anti-DN efficacy of Schisandra chinensis Fruit Mixture (SM); however, a comprehensive chemical fingerprint is still uncertain, and its mechanism of action, especially the potential therapeutic targets of anti-DN, needs to be further elucidated.

Objective: Potential mechanisms of SM action on DN were explored through network pharmacology and experimental validation.

Methods: The chemical composition of SM was analyzed using UPLC-ESI-MS/MS technology. Active bioactive components and potential targets of SM were identified using TCMSP, SwissDrugDesign, and SymMap platforms. Differentially expressed genes were determined using microarray gene data from the GSE30528 dataset. Related genes for DN were obtained from online databases, which include GeneCards, OMIM and DisGeNET. PPI networks and compound-target-pathway networks were constructed using Cytoscape. Functional annotation was performed using R software for GO enrichment and KEGG pathway analysis. The DN model was built for experimental validation using a high-sugar and high-fat diet combined with STZ induction. Hub targets and critical signaling pathways were detected using qPCR, Western blotting and immunofluorescence.

Results: Utilizing the UPLC-ESI-MS/MS coupling technique, a comprehensive analysis identified 1281 chemical components of SM's ethanol extract, with 349 of these components recognized as potential bioactive compounds through network pharmacology. Through this analysis, 126 shared targets and 15 HUB targets were pinpointed. Of these, JAK2 is regarded as the most critical gene. Enrichment analysis revealed that SM primarily operates within the PI3K/AKT signaling pathway. experiments confirmed that SM improved pathological injury and renal function in rats with DN while improving mitochondrial morphology and function and modulating the expression of proteins linked to apoptosis (cleaved-caspase-3, Bax, and Bcl-2) and pro-inflammatory factors (IL-6 and TNF-α). Mechanistically, SM alleviates DN primarily by suppressing the PI3K/AKT/mTOR and JAK2/STAT3 signaling pathways to fulfill the energy needs of renal tissues. Furthermore, molecular docking analysis provided direct validation of these findings.

Conclusion: The findings of this study offer initial indications of the active component and robust anti-inflammatory and anti-apoptotic characteristics of SM in the mitigation of DN, along with its capacity to safeguard the integrity and functionality of mitochondria. This research unequivocally validates the favorable anti-DN effects of SM, indicating its potential as a viable pharmaceutical agent for the management of DN.