Selective sodium-glucose cotransporter 2 (SGLT2) inhibitors have shown cardiovascular protection in type 2 diabetes patients with established cardiovascular (CV) disease independently of glycemic control. Although blood-derived coagulation factors are major contributors of CV diseases, their impact on the expression of SGLT1 and 2 in endothelial cells (ECs) has not been studied. This study investigated whether thrombin affects the expression of SGLT1 and 2 and examined their role in the induction of endothelial senescence.
ECs were isolated from porcine atrial tissue (AECs) and used at first passage. Endothelial senescence was assessed by determining senescence-associated beta-galactosidase (SA-beta-gal) activity, oxidative stress using dihydroethidium, and protein level by Western blot analysis and immunofluorescence staining.
Exposure of AECs to thrombin (1 or 3 U/ml) for 24 h induced an upregulation of the low protein expression level of SGLT1 and SGLT2. Thrombin induced a sustained pro-oxidant response and increased the level of SA-beta-gal activity that were both inhibited by antioxidants, an angiotensin-converting enzyme inhibitor and an AT1 receptor antagonist and also by sotagliflozin (a dual SGLT1 and SGLT2 inhibitor) and by empagliflozin (a selective SGLT2 inhibitor). In addition, thrombin upregulated the expression level of angiotensin-converting enzyme and AT1 receptors, VCAM-1 and down-regulated that of endothelial NO synthase.
In conclusion, thrombin up-regulates both SGLT1 and SGLT2 expression in AECs to sustain oxidative stress leading to endothelial senescence and dysfunction. The fact that the AT1R/NADPH oxidase/SGLT1 and 2 pathway acts in a feedforward manner suggests that inhibition of SGLT1 and/or SGLT2 appears as an attractive strategy to perpetuate the protective endothelial function on the cardiovascular system.
H. Hasan: None. S. Park: None. E. Belcastro: None. C. Auger: None. H. Lee: None. V. Schini-Kerth: Research Support; Self; Boehringer Ingelheim International GmbH.