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Ezetimibe reduces intimal hyperplasia in rabbit jugular vein graft

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1 Ezetimibe reduces intimal hyperplasia in rabbit jugular vein graft
Takashi Maekawa, MD, PhD, Kimihiro Komori, MD, PhD, Kouichi Morisaki, MD, PhD, Takeo Itoh, PhD  Journal of Vascular Surgery  Volume 56, Issue 6, Pages (December 2012) DOI: /j.jvs Copyright © 2012 Society for Vascular Surgery Terms and Conditions

2 Fig 1 Effects of ezetimibe are shown on the development of intimal hyperplasia in autologous vein grafts at 28 days after operation. A, Microscopic findings in the middle portion of the vein graft from the control group (left panel) and the ezetimibe group (right panel; elastica van Gieson staining [original magnification, ×20]). B, The arrowheads indicate the internal elastic lamina (elastica van Gieson staining [original magnification, × 400]). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

3 Fig 2 Effects of ezetimibe on proliferative activity and apoptosis in neointima of autologous vein grafts at 14 days after operation. A, Representative Ki-67 staining in the middle portion of the vein graft from nontreated control rabbits (left panel) or rabbits treated with ezetimibe (right panel; original magnification, × 400). The arrowheads indicate Ki-67–positive cells in the neointima. B, Graph shows Ki-67 index in vein grafts from nontreated control rabbits or rabbits treated with ezetimibe (n = 7 in each group). C, Representative photomicrographs stained with terminal deoxynucleotidyl transferase-mediated deoxyuride-5'-triphosphate nick-end labeling (TUNEL) in the middle portion of the vein graft from nontreated control rabbits (left panel) or rabbits treated with ezetimibe (right panel; original magnification, × 400). The arrowheads indicate TUNEL-positive cells in the neointima. D, The TUNEL index is shown in vein grafts from nontreated control rabbits or rabbits treated with ezetimibe. Results are expressed as mean ± standard error of the mean (error bars). *P < .05 vs control. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

4 Fig 3 Effects of ezetimibe on high K+-induced contraction and acetylcholine (ACh)-induced relaxation in autologous vein grafts and the effects on NOC-7–induced relaxation in endothelium-denuded vein grafts. A, After recording of the mechanical responses induced by high K+ (128 mM) and acetylcholine (10 nM to 3 μM), Nω-nitro-L-arginine (l-NNA) was applied for 60 minutes, and high-K+ and ACh-induced responses were again obtained in the presence of l-NNA in endothelium-intact strips from nontreated control rabbits (a) or rabbits treated with ezetimibe (a2). Acetylcholine, at concentrations of (1) 10 nM, (2) 30 nM, (3) 0.1 μM, (4) 0.3 μM, (5) 1 μM, and (6) 3 μM, was applied during a contraction induced by prostaglandin F2α (PGF2α). B, The absolute tension induced by 128 mM K+ before and after application of l-NNA in the vein graft from nontreated control rabbits (n = 7) or rabbits treated with ezetimibe (n = 7). C, Summary of the effects of ezetimibe on ACh-induced responses. The tonic contraction induced by PGF2α before application of acetylcholine was normalized as the relative tension of 1.0 for each strip (n = 8, control group; n = 6, ezetimibe group). l-NNA(–), Before application of l-NNA; l-NNA(+), after application of l-NNA. Results are expressed as mean ± standard error of the mean. ***P < .001 l-NNA(–) in ezetimibe vs l-NNA(+) in ezetimibe; †††P < .001 l-NNA(–) in control vs l-NNA(–) in ezetimibe. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

5 Fig 4 Effects of ezetimibe on acetylcholine (ACh)-induced increase in intracellular [Ca2+]i in endothelial cells of vein grafts. A, Fluorescence ratio (F340/F380) images of the Ca2+-sensitive dye Fura 2 in endothelial cells in the control group (left panel) and in the ezetimibe group (right panel). B, Effects on [Ca2+]i: (b1) the two groups did not differ significantly in basal [Ca2+]i; (b2) concentration-dependent effect of ACh 10 nM to 3 μM on endothelial [Ca2+]i. The change in [Ca2+]i is expressed as ΔFura 2 ratio (F340/F380) (n = 5, control group; n = 8, ezetimibe group). Data are shown as mean ± standard error of the mean. ***P < .001 vs control. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

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