Germacrone inhibits podocyte pyroptosis in diabetic kidney disease by modulating ABCA1 expression

Diabetic kidney disease (DKD) is a serious microvascular complication of diabetes with limited treatment strategies. Germacrone (Ger) has renoprotective properties, but its molecular targets and mechanisms remain unclear. This study investigated whether Ger protects against DKD by modulating podocyte pyroptosis and explored the underlying mechanism. In vivo, diabetic db/db mice were treated with Ger, with benazepril as a positive control. The effects of Ger on DKD were evaluated by assessing biochemical indices and histopathology. Molecular docking, cellular thermal shift assay (CETSA), and drug affinity responsive target stability (DARTS) were employed to assess potential target engagement between Ger and ATP-binding cassette transporter A1 (ABCA1). In vitro, ABCA1 was knocked down in high glucose (HG)-stimulated MPC5 cells, and the effects of Ger were evaluated using the Cell Counting Kit-8, enzyme-linked immunosorbent assay, flow cytometry, and Western blot. Ger reduced fasting blood glucose, improved renal function and alleviated renal pathological injury in db/db mice. Molecular docking, CETSA, and DARTS revealed potential target engagement between Ger and ABCA1. Ger increased ABCA1 mRNA and protein levels, enhanced cholesterol efflux, reduced proinflammatory factor release, and suppressed pyroptosis in db/db mice and HG-stimulated MPC5 cells. ABCA1 knockdown weakened the protective effects of Ger, whereas toll-like receptor 4 (TLR4) inhibition partially reversed the adverse effects of ABCA1 knockdown. Ger alleviates DKD-associated podocyte injury by regulating ABCA1 expression and TLR4-associated pyroptosis, supporting its further evaluation as a therapeutic candidate for DKD.