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

Lin Fu,1 Nenghua Zhang,1 Xingying Chen,1 Xiuqin Xu,2 Yunqiu Shen1 1Department of Clinical Laboratory, Jiaxing Hospital of Traditional Chinese Medicine, Jiaxing, People’s Republic of China; 2Department of Nephrology, Jiaxing Hospital of Traditional Chinese Medicine, Jiaxing, People’s Republic of ChinaCorrespondence: Lin Fu, Email linfu0725@163.comPurpose: 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.Keywords: membranous nephropathy, Buyang Huanwu decoction, network pharmacology, AGE/RAGE pathway