PHARMACOLOGICAL EFFECTS AND MECHANISMS OF DA-YU-GONG DECOCTION IN THE TREATMENT OF MESANGIAL PROLIFERATIVE GLOMERULONEPHRITIS

Mesangial proliferative glomerulonephritis (MsPGN) is a common and heterogeneous form of chronic kidney disease, characterized by excessive mesangial cell proliferation, inflammation, and extracellular matrix deposition. The clinical complexity and inter-individual variability in underlying mechanisms pose challenges to effective treatment, making the development of precision medicine strategies essential. Da-Yu-Gong Decoction (DYGD), a traditional Chinese medicine formula comprising Rhei Radix et Rhizoma and Rosae Multiflorae Fructus, has been clinically applied in MsPGN, yet its mechanistic basis remains unexplored. This study aimed to elucidate the pharmacological effects and molecular mechanisms of DYGD, providing a foundation for mechanism-driven, precision interventions in MsPGN.

DYGD was extracted with water and fractionated; the ethyl acetate fraction (DYGDE) was selected based on its anti-proliferative action on mesangial cells for further study. It was analyzed by UPLC to ensure reproducibility. Anti-MsPGN effects were evaluated in PDGF-BB-stimulated human (HMC) and rat (RMC) mesangial cells, as well as in a rat MsPGN model induced by anti-Thy1 antibody. Key outcomes included cell proliferation, molecular marker expression, renal function, and histopathology. Network pharmacology was employed to predict potential active compounds and molecular targets of DYGDE. Compound–target interactions were assessed by molecular docking (AutoDock Vina), and binding stability was confirmed via molecular dynamics simulations. This integrative approach facilitated precise validation of compound–target interactions, supporting mechanism-based strategies for MsPGN.

UPLC analysis established the chemical profile of DYGDE, with quantification of marker compounds. In vitro, DYGDE significantly inhibited PDGF-BB–induced mesangial cell proliferation and downregulated mRNA and protein expression of proliferation markers (cyclin D1, CDK4, PCNA), inflammatory mediators (ICAM-1, VCAM-1), and fibronectin. In vivo, DYGDE ameliorated renal injury in MsPGN rats, as evidenced by reduced 24-hour proteinuria, urine protein-to-creatinine ratio, glomerular cell proliferation, fibronectin deposition, and renal inflammation. Network pharmacology highlighted the RAS/MAPK signaling pathway as a major target of DYGDE. DYGDE reversed RAS activation and decreased phosphorylation of c-Raf, MEK, and ERK in both anti-Thy1-induced MsPGN rats and PDGF-BB-stimulated mesangial cells. The anti-proliferative effects of DYGDE were abolished by the RAS activator SOS1 agonist-1, confirming its action through the RAS/MAPK pathway. Toralactone, rheosmin, and physcion were identified as key active components mediating RAS/MAPK pathway inhibition. Molecular docking demonstrated that toralactone exhibited the strongest binding affinity to PDGFR, an upstream regulator of RAS/MAPK signaling (binding free energy: -10.2 kcal/mol), with molecular dynamics simulations confirming high binding stability. Collectively, these findings elucidate the mechanism of DYGDE and underscore its potential for mechanism-based, patient-tailored therapy in MsPGN.

DYGDE exerts anti-MsPGN effects primarily through targeted inhibition of the RAS/MAPK signaling pathway. Identification of toralactone as a key bioactive component interacting with PDGFR provides a strong mechanistic rationale for precision therapeutic strategies; however, this interaction requires further experimental validation. These results position DYGDE—and specifically toralactone—as a promising candidate for personalized precision medicine in MsPGN, offering novel perspectives for TCM-based interventions in this disease. The following is the graphical abstract of this work.