The Sodium-Glucose Cotransporter-2 Inhibitor Canagliflozin Alleviates Endothelial Dysfunction Following In Vitro Vascular Ischemia/Reperfusion Injury in Rats

Abstract

Vascular ischemia/reperfusion injury (IRI) contributes to graft failure and adverse clinical outcomes following coronary artery bypass grafting. Sodium-glucose-cotransporter (SGLT)-2-inhibitors have been shown to protect against myocardial IRI, irrespective of diabetes. We hypothesized that adding canagliflozin (CANA) (an SGLT-2-inhibitor) to saline protects vascular grafts from IRI. Aortic rings from non-diabetic rats were isolated and immediately mounted in organ bath chambers (control, n = 9–10 rats) or underwent cold ischemic preservation in saline, supplemented either with a DMSO vehicle (IR, n = 8–10 rats) or 50µM CANA (IR + CANA, n = 9–11 rats). Vascular function was measured, the expression of 88 genes using PCR-array was analyzed, and feature selection using machine learning was applied. Impaired maximal vasorelaxation to acetylcholine in the IR-group compared to controls was significantly ameliorated by CANA (IR 31.7 ± 3.2% vs. IR + CANA 51.9 ± 2.5%, p < 0.05). IR altered the expression of 17 genes. Ccl2, Ccl3, Ccl4, CxCr4, Fos, Icam1, Il10, Il1a and Il1b have been found to have the highest interaction. Compared to controls, IR significantly upregulated the mRNA expressions of Il1a and Il6, which were reduced by 1.5- and 1.75-fold with CANA, respectively. CANA significantly prevented the upregulation of Cd40, downregulated NoxO1 gene expression, decreased ICAM-1 and nitrotyrosine, and increased PECAM-1 immunoreactivity. CANA alleviates endothelial dysfunction following IRI.

Graft Preservation Solution DuraGraft® Alleviates Vascular Dysfunction Following In Vitro Ischemia/Reperfusion Injury in Rats

“…To extrude oxygen, saline or DuraGraft ® solutions were gassed with nitrogen. As previously described [15], the aortic rings were placed in test tubes containing 0.9% saline (IR group, n = 28-35 rings, 7 rats) or DuraGraft ® (IR+DuraGraft ® group (n = 36 rings, 9 rats), and stored for 24 h at 4 • C. Following cold ischemic conservation, the rings were mounted in organ baths. To mimic free radical generation and endothelial dysfunction, as occur during reperfusion and re-oxygenation in vivo, 200 µM sodium hypochlorite was added to the organ chambers for 30 min.…”

“…Aortic rings were mounted on a stainless-steel hook in organ chambers containing 30 ml of KHL and continuously bubbled with 95% O 2 and 5% CO 2 at 37 • C and pH 7.4 (EMKA Technologies S.A.S, Paris, France). As previously reported [15], the tissue was initially equilibrated for 20 min at a resting tension before any experimental intervention. During an additional 60 min of equilibration, the passive tension was adjusted periodically to 2 g, during which the baths were rinsed with fresh KHL every 30 min, a precaution against interfering metabolites.…”

“…As previously described [15], immunohistochemistry was performed on buffered paraformaldehyde solution (4%) fixed, paraffin embedded, distal regions of the aortic segments. Four µm thick sections were cut with the Leica microtome (Leica Biosystems Nussloch GmbH, Nussloch, Germany) and placed on slides.…”

Cardiovascular Benefits from Gliflozins: Effects on Endothelial Function

“…A very recent investigation documented that the addition of canagliflozin to the storage solution elicited a downregulation of proinflammatory genes and their products in aortic rings from nondiabetic rats following an in vitro ischemia/reperfusion (I/R) injury [92].…”

Scutellarin Reduces Cerebral Ischemia Reperfusion Injury Involving in Vascular Endothelium Protection and PKG Signal

“…It can trigger tissue injury caused by various pathophysiological mechanisms. IR directly affects the vascular wall and luminal surface of blood vessels and cause damage such as hemorrhage, capillary plugging, adhesion and infiltration of granulocytes, impaired vascular permeability, and endothelial dysfunction (ED) [ 3 – 5 ]. Endothelial cells which are particularly susceptible to IR injury play an active role in the case of IR-induced organ damage [ 6 ].…”