Inhibitory Mechanism of Buyang Huanwu Decoction on AGE/RAGE Pathway in Membranous Nephropathy: Integration of Network Pharmacology and Cell Model Validation

Purpose: The Buyang Huanwu Decoction (BYHWD) has demonstrated therapeutic potential in renal-related disorders; however, its pharmacological mechanisms are still poorly understood. Therefore, the aim of this study was to elucidate the regulatory mechanisms of BYHWD in membranous nephropathy (MN). Methods: Network pharmacology was used to identify BYHWD-related target genes for MN. Enrichment analyses were conducted to determine the relevant biological functions and signaling pathways. An integrated "compound-target-pathway" interaction network was established. The binding affinities between the active compounds and target proteins were determined via molecular docking. Two podocyte injury models were established using zymosan-activated serum (ZAS)-induced MPC-5 cells and Angiotensin II (Ang II)-induced AB8/13 cells. Cell viability was assessed using the Cell Counting Kit-8 (CCK-8) assay. ELISA was used to quantify the levels of pro-inflammatory cytokines, membrane attack complexes (MAC, C5b-9), and advanced glycation end products (AGE), while Western blotting was performed to determine receptor for advanced glycation end products (RAGE) protein expression. Results: BYHWD shared 230 genes with the MN-related targets. GO analysis indicated its involvement in regulating cell proliferation, apoptosis, and inflammation. KEGG analysis highlighted the modulation of the AGE-RAGE signaling pathway. IL-1β showed the highest diagnostic value in the machine learning analysis. Molecular docking revealed stable interactions between key compounds (myristic acid, stigmasterol, quercetin, and β-sitosterol) and target proteins. Both ZAS and Ang II inhibited podocyte proliferation and increased the levels of pro-inflammatory cytokines and C5b-9, whereas BYHWD reversed these effects. It also suppressed AGE and RAGE expression, and these effects were counteracted by pathway agonists. Conclusion: BYHWD may improve podocyte injury by inhibiting AGE/RAGE and suppressing inflammatory responses and complement activation, providing a preliminary basis for its clinical application.