The Combined Administration of Disulfiram and Baicalin Alleviates Membranous Nephropathy by Inhibiting Podocyte Inflammation and Pyroptosis

BACKGROUND: Membranous nephropathy (MN) is an autoimmune kidney disease, and its pathogenesis is related to inflammation and podocyte injury. Both disulfiram and baicalin can relieve the symptoms of MN through anti-inflammatory effects. However, it is still unclear whether disulfiram and baicalin can enhance the therapeutic effect of MN by combination therapy. METHODS: We screened the optimal drug concentration of disulfiram and baicalin for incubation of AB8/13 cells by CCK-8 assay. Angiotensin II was used to stimulate AB8/13 cells to construct a MN cell model. Subsequently, cell proliferation assays, network pharmacology analysis, ELISA assay, and pyroptosis detection were employed to investigate the effects of disulfiram combined with baicalin in MN. RESULTS: Firstly, 250 nM disulfiram and 5 μmol/L baicalin were selected as the optimal dose for the treatment of MN. Cell proliferation assay showed that the combined administration of disulfiram and baicalin could reduce Angiotensin II-induced podocyte injury. Subsequently, a total of 129 genes were identified as potential therapeutic targets of the combination therapy involving disulfiram and baicalin for MN. These targets mainly focused on regulating inflammatory-related functions and pathways. Disulfiram combined with baicalin inhibited the inflammation of MN by suppressing the levels of TNF-α, IL-8 and C-reactive protein and promoting the levels of TGF-β and IL-10. Disulfiram combined with baicalin significantly attenuated Angiotensin II-induced podocyte pyroptosis, as indicated by reduction of Caspase-1 and GSDMD-N, lactate dehydrogenase release and the percentage of podocytes stained with propidium iodide. CONCLUSION: Compared with monotherapy using either disulfiram or baicalin alone for MN, the combination has a protective role in podocyte injury by inhibiting inflammation and pyroptosis. This discovery provides a new perspective for clarifying the pharmacological effects of disulfiram and baicalin to podocyte injury in MN.