Inhibition of Caspase-3/GSDME Pathway-Mediated Pyroptosis of Renal Tubular Epithelial Cells by Dagliflozin in the Pathogenesis of Diabetic Kidney Disease and Study of Its Mechanism

Objective: Investigating the effects and mechanisms of dapagliflozin on Pyroptosis of renal tubular epithelial cells under high-glucose conditions through the regulation of the Caspase-3/GSDME signaling pathway, providing experimental evidence for the clinical treatment of diabetic kidney disease.

Methods: Human renal tubular epithelial cells (HK-2) were cultured in vitro and divided into control group (5mmol/L D-glucose), high-glucose group (30mmol/L D-glucose), dagliflozin group (2.5μmol/L dagliflozin), and monoinhibitor group (20μmol/L Caspase-1 inhibitor), dual inhibitor group (20μmol/L Caspase-1 inhibitor + 50μmol/L Caspase-3 inhibitor), and SiRNA transfection group. All groups were intervened for 48h. The cell viability was detected by cell counting kit-8 and the glucose and dagliflozin concentrations of the intervention were determined. Caspase-1, Caspase-3, GSDMD, GSDME, GSDME-N, Caspase-8, NF-κB were detected by Western blot. Detection of cellular Pyroptosis in each group by flow cytometry.

Results: Compared with the control group, the D-glucose group showed decreased cell viability, increased cell Pyroptosis, and increased levels of Caspase-1, Caspase-3, GSDMD, GSDME, GSDME-N, Caspase-8, NF-κB, and Other related proteins (P<0.05). Compared with the D-glucose group, the rate of cellular Pyroptosis and the levels of Caspase-1, Caspase-3, GSDMD, GSDME, GSDME-N and Other related proteins were decreased in the dagliflozin group and the dual inhibitor group (P<0.05). Compared with the transfected control group, the cellular Pyroptosis rate and the levels of Caspase-1, Caspase-3, GSDMD, GSDME, GSDME-N, and Other related proteins were then further reduced in the transfected group targeting SGLT2 knockdown (P<0.05).

Conclusion: In the proximal tubular cells of diabetic kidney disease, dagliflozin inhibited high glucose-induced Pyroptosis of human HK-2, and its mechanism of action may be related to the inhibition of Caspase 3/GSDME pathway signaling.

dagliflozin; diabetic kidney disease; pyroptosis; renal tubular epithelial cells.