Background and aims: Circulating microparticles (MPs) from patients with coronary artery diseases (CAD) have been shown to promote endothelial senescence and dysfunction involving the pro-oxidant local angiotensin system. Sodium-glucose cotransporters (SGLTs)2 inhibitors decreased the risk of cardiovascular disease in patients with type 2 diabetes and this effect appears to be independent of glycemic control. Moreover, high glucose and H2O2has been shown to cause a redox-sensitive upregulation of SGLT1 and 2 in coronary artery endothelial cells (ECs). Therefore, this study examined whether CAD MPs and angiotensin II (Ang II, a potent NADPH oxidase-dependent inducer of oxidative stress) stimulate SGLT1 and 2 expression in ECs, and assessed their role in the induction of endothelial dysfunction. Materials and methods: ECs were isolated from porcine coronary arteries. The protein expression level was assessed by Western blot analysis and immunocytochemical staining, oxidative stress using dihydroethidium staining, and senescence by senescence-associated beta-galactosidase activity (SA-betagal activity). Circulating CAD MPs were collected from blood samples of patients (50-83 years) with established cardiovascular disease. Results: ECs expressed low levels of SGLT1 and SGLT2 proteins. Exposure of ECs to CAD MPs (10 nM PhtdSer eq) increased the SGLT1 and SGLT2 protein levels in 3/5 patients. Exposure of ECs to Ang II caused a time- and concentration-dependent increase in the protein levels of SGLT1 and SGLT2, with a significant increase observed at concentrations as low as 10 nM. An increased SGLT1 and SGLT2 immunofluorescence signal was also observed in response to Ang II and H2O2. Ang II increased the level of oxidative stress, SA-beta-gal activity, senescence markers (p53, p21 and p16), VCAM-1, MCP-1, tissue factor (TF) and the local angiotensin system (ACE, AT1R), and down-regulated that of eNOS. CAD MPs decreased eNOS level in 4/5 patients and increased VCAM-1 level in 5/5 patients. The CAD MPs- and the Ang II-induced expression of SGLT 1 and 2, VCAM-1 and down-regulation of eNOS were prevented by the NADPH oxidase inhibitor VAS-2870, the AT1R antagonist losartan, and by the dual SGLT1/2 inhibitor LX-4211 and the selective SGLT2 inhibitor, empagliflozin. Conclusion: The present findings indicate that CAD MPs and Ang II upregulate the expression of SGLT1 and SGLT2 protein levels in ECs, and that they promote endothelial dysfunction. They further suggest that inhibition of SGLT1 and/or SGLT2 might be an attractive strategy to protect the arterial wall and, hence, the development of cardiovascular diseases. Supported by: Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany Disclosure: S. Park: Grants; Unrestricted research grant from Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
Coronary artery disease patient microparticles up-regulate sodium-glucose cotransporters 1 and 2 expression to promote atherosclerotic responses in endothelial cells
Belcastro, E;
2019-01-01
Abstract
Background and aims: Circulating microparticles (MPs) from patients with coronary artery diseases (CAD) have been shown to promote endothelial senescence and dysfunction involving the pro-oxidant local angiotensin system. Sodium-glucose cotransporters (SGLTs)2 inhibitors decreased the risk of cardiovascular disease in patients with type 2 diabetes and this effect appears to be independent of glycemic control. Moreover, high glucose and H2O2has been shown to cause a redox-sensitive upregulation of SGLT1 and 2 in coronary artery endothelial cells (ECs). Therefore, this study examined whether CAD MPs and angiotensin II (Ang II, a potent NADPH oxidase-dependent inducer of oxidative stress) stimulate SGLT1 and 2 expression in ECs, and assessed their role in the induction of endothelial dysfunction. Materials and methods: ECs were isolated from porcine coronary arteries. The protein expression level was assessed by Western blot analysis and immunocytochemical staining, oxidative stress using dihydroethidium staining, and senescence by senescence-associated beta-galactosidase activity (SA-betagal activity). Circulating CAD MPs were collected from blood samples of patients (50-83 years) with established cardiovascular disease. Results: ECs expressed low levels of SGLT1 and SGLT2 proteins. Exposure of ECs to CAD MPs (10 nM PhtdSer eq) increased the SGLT1 and SGLT2 protein levels in 3/5 patients. Exposure of ECs to Ang II caused a time- and concentration-dependent increase in the protein levels of SGLT1 and SGLT2, with a significant increase observed at concentrations as low as 10 nM. An increased SGLT1 and SGLT2 immunofluorescence signal was also observed in response to Ang II and H2O2. Ang II increased the level of oxidative stress, SA-beta-gal activity, senescence markers (p53, p21 and p16), VCAM-1, MCP-1, tissue factor (TF) and the local angiotensin system (ACE, AT1R), and down-regulated that of eNOS. CAD MPs decreased eNOS level in 4/5 patients and increased VCAM-1 level in 5/5 patients. The CAD MPs- and the Ang II-induced expression of SGLT 1 and 2, VCAM-1 and down-regulation of eNOS were prevented by the NADPH oxidase inhibitor VAS-2870, the AT1R antagonist losartan, and by the dual SGLT1/2 inhibitor LX-4211 and the selective SGLT2 inhibitor, empagliflozin. Conclusion: The present findings indicate that CAD MPs and Ang II upregulate the expression of SGLT1 and SGLT2 protein levels in ECs, and that they promote endothelial dysfunction. They further suggest that inhibition of SGLT1 and/or SGLT2 might be an attractive strategy to protect the arterial wall and, hence, the development of cardiovascular diseases. Supported by: Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany Disclosure: S. Park: Grants; Unrestricted research grant from Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
