Dapagliflozin Inhibits Thrombin Induced ADAM17 Phosphorylation and Downstream ERK/p-c-Jun-mediated CTGF Induction to...

Vascular restenosis following angioplasty remains a major clinical challenge driven by excessive vascular smooth muscle cell (VSMC) migration, proliferation, and fibrosis. Thrombin, a potent signaling molecule beyond its coagulative function, promotes neointimal hyperplasia. Dapagliflozin, a sodium-glucose cotransporter 2 (SGLT2) inhibitor, has been shown to exert cardiovascular protection independent of its glucose-lowering effects, yet its role in thrombin-mediated fibrotic signaling remains unclear.

Cultured A10 vascular smooth muscle cells were used to evaluate thrombin- stimulated A Disintegrin And Metalloprotease 17 (ADAM17), ERK1/2, c-Jun, and connective tissue growth factor (CTGF) by immunoblotting and migration analyses. Enzymatic activity (ADAM17 and thrombin) assays were tested by cell-free system kit. Pharmacological inhibition of ADAM17 by TAPI-1 and extracellular signal-regulated kinase (ERK) by U0126 were employed to delineate pathway hierarchy. An in vivo rat carotid artery balloon injury model was used to assess the effects of dapagliflozin on neointimal formation.

Thrombin stimulation markedly increased ADAM17 phosphorylation, ERK1/2 and c-Jun activation, and CTGF expression in A10 cells. Dapagliflozin significantly suppressed these responses in a concentration-dependent manner without affecting thrombin enzymatic activity. Inhibition of ADAM17 recapitulated the effects of dapagliflozin, confirming ADAM17 as a key upstream mediator of thrombin-induced ERK/c-Jun/CTGF signaling. Functionally, dapagliflozin attenuated VSMC migration in vitro and significantly reduced neointimal hyperplasia in the rat carotid artery balloon injury model.

Dapagliflozin inhibits thrombin-induced ADAM17 activation and downstream ERK1/2-c-Jun-CTGF signaling, thereby limiting VSMC migration and neointimal formation after vascular injury. This SGLT2-independent mechanism provides new insight into the pleiotropic vasculoprotective actions of dapagliflozin and identifies ADAM17 as a potential therapeutic target for preventing vascular restenosis and fibrosis-related cardiovascular disease.