Endocrine Abstracts

Introduction: Treatment with sodium glucose co-transporter2(SGLT2) inhibitors was found to reduce the incidence of cardiovascular events in diabetic patients.Herein, we investigated the effects of long-term treatment with canagliflozin on atherosclerosis development in the aorta of Apolipoprotein E knockout (Apo-E^(−/−)) mice as well as on biochemical and immunohistochemical markers related to atherosclerosis.

Methods: At 4 weeks of age, mice were switched from normal to a high-fat diet. At 8 weeks of age, Apo-E^(−/−) mice were divided into control-group (6 mice) treated with 0.5%hydroxypropyl methylcellulose per os (po.) and Cana-group (6 mice) treated with canagliflozin (10 mg/Kgr d., po.). After 5 weeks, animals were sacrificed and heart and aorta were removed. Sections stained with hematoxylin-eosin (H&E) were used for histomorphometry whereas Masson’s stained tissues were used to quantify the collagen content. Aorta root sections were stained for MCP-1, CD68, a-smooth muscle actin, MMP-2, MMP-9. q-PCR experiments were carried out to quantify the mRNA expression of MMP-2, MMP-9 their inhibitors TIMP-1 and TIMP-2, IL-6, VCAM-1, ICAM-1 and MCP-1 in the aorta.

Results: Cana-group was found to have lowered LDL-cholesterol, triglycerides and glucose levels (P<0.01) compared to control group. Heart rate was lowered in Cana-group compared to control-group (P<0.05). Histomorphometry analysis revealed that one out of six mice of Cana-group vs four out of six mice in control group developed atheromatosis, while the plaque area in aortic root was significantly lower and collagen was two times more intense in Cana-group compared to control. Immunohistochemistry showed that MCP-1, a-actin and CD68 were less expressed in aortic root of Cana-group compared to control, whereas MMP-2 expression was more intense. VCAM-1 mRNA levels were lower while TIMP-1 expression was significantly higher in Cana-group compared to control. No significant differences in MMP-9, MMP-2, TIMP-2 ICAM-1 and IL-6 mRNA levels were observed between two groups. There was a decrease in MCP-1 and increase in MMP-9 mRNA expression in Cana-group compared to control, however not statistically significant.

Conclusion: Preclinical data revealed that long-term administration of Canagliflozin attenuates the progression of atherosclerosis via reducing i) hyperlipidemia and hyperglycemia, ii) inflammatory process, by lowering the expression of inflammatory molecules such as MCP-1 and VCAM-1. Moreover, Canagliflozin was found to increase the stability of atheromatous plaque through increasing the expression of TIMP-1 (MMP-2-inhibitor).