Yishen Quzhuo decoction improve chronic kidney disease by regulating amino acid metabolism: An experimental validation guided by network pharmacology

Introduction: YiShen QuZhuo Decoction (YSQZD) is a clinically proven traditional Chinese medicine formula for chronic kidney disease (CKD), but its underlying mechanism lacks systematic investigation. CKD is a prevalent renal disorder with limited effective treatments, and traditional herbal medicines are gaining attention as safer alternatives. Renal tubular injury is a key pathological change in CKD, closely linked to renal interstitial fibrosis. Methods: Network pharmacology was used to predict active ingredients, key targets, and potential mechanisms of YSQZD against CKD. BATMAN-TCM, GeneCards, STRING, and DAVID databases were employed for target screening, PPI network construction, and GO/KEGG enrichment analysis. TGF-β1-induced HK-2 cell injury model was used for in vitro validation. MTT assay, immunofluorescence staining, and GC-MS-based amino acid metabolomics were performed to evaluate the efficacy and mechanism of YSQZD. Conclusion: Network pharmacology identified 19 active ingredients (score >100, based on BATMAN-TCM default high-confidence threshold and previous literature support) and 57 common targets of YSQZD and CKD. GO/KEGG analysis revealed 26 signaling pathways, including 5 amino acid metabolism-related pathways (P<0.05). In vitro experiments (n=6 biological replicates per group) showed YSQZD dose-dependently alleviated TGF-β1-induced EMT in HK-2 cells. Western blot confirmed upregulated E-cadherin and downregulated α-SMA. Amino acid metabolomics indicated YSQZD significantly regulated aspartic acid, glutamine, and leucine levels (P < 0.05). Discussion: YSQZD exerts anti-CKD effects through multi-component, multi-target, and multi-pathway mechanisms. It regulates amino acid metabolism (especially aspartic acid) and inhibits EMT, which may be associated with SLC family transporters. The findings provide a scientific basis for YSQZD's clinical application, but further in vivo and clinical studies are needed to confirm its efficacy.