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.