Angiotensin II-induced upregulation of SGLT1 and 2 contributes to human microparticle\u2010stimulated endothelial senescence and dysfunction: protective effect of gliflozins

Eugenia Belcastro,Patrick Ohlmann,Benjamin Marchandot,Florence Toti,Malak Abbas,Kensuke Matsushita,Laurence Jesel,Cyril Auger,Sin-Hee Park,Valérie B Schini-Kerth,Olivier Morel,Hira Hasan

doi:10.1186/s12933-021-01252-3

Abstract

BackgroundSodium-glucose cotransporter 2 (SGLT2) inhibitors reduced cardiovascular risk in type 2 diabetes patients independently of glycemic control. Although angiotensin II (Ang II) and blood-derived microparticles are major mediators of cardiovascular disease, their impact on SGLT1 and 2 expression and function in endothelial cells (ECs) and isolated arteries remains unclear.Methods ECs were isolated from porcine coronary arteries, and arterial segments from rats. The protein expression level was assessed by Western blot analysis and immunofluorescence staining, mRNA levels by RT-PCR, oxidative stress using dihydroethidium, nitric oxide using DAF-FM diacetate, senescence by senescence-associated beta-galactosidase activity, and platelet aggregation by aggregometer. Microparticles were collected from blood of patients with coronary artery disease (CAD-MPs).ResultsAng II up-regulated SGLT1 and 2 protein levels in ECs, and caused a sustained extracellular glucose- and Na+-dependent pro-oxidant response that was inhibited by the NADPH oxidase inhibitor VAS-2780, the AT1R antagonist losartan, sotagliflozin (Sota, SGLT1 and SGLT2 inhibitor), and empagliflozin (Empa, SGLT2 inhibitor). Ang II increased senescence-associated beta-galactosidase activity and markers, VCAM-1, MCP-1, tissue factor, ACE, and AT1R, and down-regulated eNOS and NO formation, which were inhibited by Sota and Empa. Increased SGLT1 and SGLT2 protein levels were observed in the rat aortic arch, and Ang II- and eNOS inhibitor-treated thoracic aorta segments, and were associated with enhanced levels of oxidative stress and prevented by VAS-2780, losartan, Sota and Empa. CAD-MPs promoted increased levels of SGLT1, SGLT2 and VCAM-1, and decreased eNOS and NO formation in ECs, which were inhibited by VAS-2780, losartan, Sota and Empa.ConclusionsAng II up-regulates SGLT1 and 2 protein expression in ECs and arterial segments to promote sustained oxidative stress, senescence and dysfunction. Such a sequence contributes to CAD-MPs-induced endothelial dysfunction. Since AT1R/NADPH oxidase/SGLT1 and 2 pathways promote endothelial dysfunction, inhibition of SGLT1 and/or 2 appears as an attractive strategy to enhance the protective endothelial function.

Highlights

Sodium-glucose cotransporter 2 (SGLT2) inhibitors reduced cardiovascular risk in type 2 diabetes patients independently of glycemic control
Data are expressed as mean ± SEM of n = 3–4. *P < 0.05 vs. control thoracic aorta (a–j, l, m) and outer aortic arch (k), and #P < 0.05 vs. Ang Angiotensin II (II)-treated thoracic aorta (a–e, l) and Nω-nitro L-arginine (L-NA)-treated thoracic aorta (f–j, m) eNOS/β-tubulin VCAM-1/β-tubulin
The major findings of the present study indicate that angiotensin II (Ang II) and MPs derived from coronary artery disease (CAD) patients cause via the AT1 receptor/NADPH oxidase pathway a redox-sensitive up-regulation of the expression of SGLT1 and 2 in endothelial cells (ECs), which, in turn, have a key role to promote endothelial senescence and dysfunction

Summary

Introduction

Sodium-glucose cotransporter 2 (SGLT2) inhibitors reduced cardiovascular risk in type 2 diabetes patients independently of glycemic control. Exposure of endothelial cells (ECs) to circulating MPs from acute coronary syndrome patients induced premature endothelial senescence and thrombogenicity through activation of the Ang II/AT1R/NADPH oxidase pathway [13]. Such findings are in good agreement with observations indicating that the angiotensin system contributes to the induction of endothelial dysfunction in the inner curvature of the aortic arch [14], in experimental models of atherosclerosis and aging, hypertension, diabetes and in patients at high cardiovascular risk [15,16,17]. ECs express angiotensin-converting enzyme (ACE) that stimulates the conversion of angiotensin I (Ang I) into the biologically active Ang II [18], which, in turn, causes NADPH oxidase-mediated oxidative stress and promotes vasoconstriction, endothelial senescence and dysfunction, and vascular and cardiac remodeling [19, 20]

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