Gooseberry, citrus peel, and licorice extracts as multi-target modulators of inflammatory and metabolic pathways in LPS-induced muscle wasting: integrative network pharmacology and computational docking analysis

Abstract Sarcopenia affects over 50 million individuals worldwide but lacks precision therapeutic strategies. The ADAMTSL3 rs4842838 (Val661Leu) variant may dysregulate TGF-β signaling and inflammatory responses in muscle wasting. This study aimed to establish an integrated computational-to-biological framework for identifying natural compounds (NCs) targeting inflammatory muscle wasting, using ADAMTSL3 rs4842838 (Val661Leu) variant-informed molecular docking and network pharmacology to prioritize candidate herbal extracts, which were subsequently validated in lipopolysaccharide (LPS)-induced C2C12 myotubes. Molecular docking of 25,000 NCs was performed against wild-type (WT) and mutant-type (MT) ADAMTSL3 Val661Leu structures. Compounds with binding energies ≤–9.5 kcal/mol were evaluated via network pharmacology. Biological validation used LPS-induced inflammatory muscle wasting in C2C12 myoblasts treated with gooseberry, licorice, and citrus peel extracts (1–125 µg/mL). Molecular clustering revealed superior performance for MT-selected NCs versus WT (silhouette score 0.75 vs. 0.63). MT-selected NCs required nitrogen-containing groups. Network pharmacology analysis identified licorice NCs targeting PDGFRB , AKT1 , mTOR , and SOD1 pathways, while gooseberry NCs modulated matrix regulation via MMP3/MMP9. LPS treatment increased MMP3 expression by 51%; gooseberry and licorice extracts normalized MMP3 levels, while citrus peel extract showed partial recovery. All extracts upregulated muscle development genes ( MYOG , MyoD1 , ADAMTSL3 ), downregulated myostatin, reduced lipid peroxidation, and suppressed inflammatory markers (IL-1β, IL-6) ( P < 0.05). The extracts enhanced TGF-β and insulin signaling while attenuating NF-κB activation, suggesting that the ADAMTSL3 rs4842838 variant disrupted TGF-β signaling and contributed to muscle wasting through dysregulated inflammatory responses. In conclusion, docking-informed network pharmacology analysis combined with in vitro validation supports a precision nutrition framework for inflammatory muscle wasting. Gooseberry, licorice, and citrus peel extracts exhibit multi-pathway muscle-protective effects through coordinated modulation of oxidative stress, inflammation, and TGF-β signaling.