DOI: /j.atherosclerosis

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DOI: 10.1016/j.atherosclerosis.2017.04.001 Liraglutide, a GLP-1 receptor agonist, inhibits vascular smooth muscle cell proliferation by enhancing AMP-activated protein kinase and cell cycle regulation, and delays atherosclerosis in ApoE deficient mice Teruo Jojima, Kohsuke Uchida, Kazumi Akimoto, Takanori Tomotsune, Kazunori Yanagi, Toshie Iijima, Kunihiro Suzuki, Kikuo Kasai, Yoshimasa Aso Atherosclerosis Volume 261, Pages 44-51 (June 2017) DOI: 10.1016/j.atherosclerosis.2017.04.001 Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 1 Inhibitory effects of liraglutide on atherosclerotic plaque lesions in ApoE−/− mice. Quantification of arteriosclerotic plaque with oil red O staining showed a significant greater lesion in ApoE −/− mice than in control mice. Treatment with both liraglutide 150 μg/day and 400 μg/day for 4 weeks suppressed the lesions of atherosclerotic plaques compared with the vehicle-treated ApoE KO mice (p < 0.05, p < 0.05, respectively). N = 5. Atherosclerosis 2017 261, 44-51DOI: (10.1016/j.atherosclerosis.2017.04.001) Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 2 Effects of liraglutide on phosphorylated AMPK expression in ApoE−/− mice. Immunohistochemical analysis of p-AMPK staining in the aorta indicated that treatment with both liraglutide 150 μg/day and 400 μg/day for 4 weeks increased the expression of p-AMPK compared with the vehicle-treated ApoE KO mice (p < 0.01, p < 0.01, respectively) A: Magnification ×100. B: Magnification ×400. (N = 6). Atherosclerosis 2017 261, 44-51DOI: (10.1016/j.atherosclerosis.2017.04.001) Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 3 Concentration-response curve to the endothelium-dependent vasodilator acetylcholine in the thoracic aorta taken from ApoE mice treated with liraglutide. Treatment with both liraglutide 150 μg/day and 400 μg/day for 4 weeks significantly improved endothelial function compared with the vehicle-treated ApoE KO mice. (p < 0.05, respectively). (N = 4). Atherosclerosis 2017 261, 44-51DOI: (10.1016/j.atherosclerosis.2017.04.001) Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 4 Effects of liraglutide on the real-time proliferation of rat VSMCs evaluated by the xCELLigence DP system. Ang II significantly augmented VSMC proliferation compared with control (p < 0.05). Treatment with 1 μM liraglutide and 1 mM AICAR inhibited Ang II-induced VSMC proliferation (p < 0.0001, respectively). (N = 4). Atherosclerosis 2017 261, 44-51DOI: (10.1016/j.atherosclerosis.2017.04.001) Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 5 Effects of liraglutide on phosphorylation of AMPK in rat VSMCs. Time course studies revealed that liraglutide stimulated phosphorylation of AMPK and its peak was shown at 15 min. The p-AMPK/total AMPK ratio was significantly higher at 15 min than at baseline (p < 0.01). Similarly, liraglutide stimulated phosphorylation of ACC. Liraglutide-induced activation of AMPK was abolished by exendin 9-39, an antagonist of GLP-1. (N = 4). Atherosclerosis 2017 261, 44-51DOI: (10.1016/j.atherosclerosis.2017.04.001) Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 6 Effect of liraglutide or AICAR on cell cycle progression in Ang II-induced rat VSMCs evaluated by flow cytometry. Treatment with liraglutide and AICAR decreased the cells in the G2/M phase of Ang II-induced rat VSMCs. Values are expressed as a percentage of each phase cell of total cells. Each value represents mean ± SE from 5 independent experiments. Atherosclerosis 2017 261, 44-51DOI: (10.1016/j.atherosclerosis.2017.04.001) Copyright © 2017 Elsevier B.V. Terms and Conditions