Sarpogrelate hydrochloride reduced intimal hyperplasia in experimental rabbit vein graft Akio Kodama, MD, Kimihiro Komori, MD, PhD, Keisuke Hattori, MD,

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Presentation transcript:

Sarpogrelate hydrochloride reduced intimal hyperplasia in experimental rabbit vein graft Akio Kodama, MD, Kimihiro Komori, MD, PhD, Keisuke Hattori, MD, PhD, Dai Yamanouchi, MD, PhD, Junko Kajikuri, PhD, Takeo Itoh, PhD Journal of Vascular Surgery Volume 49, Issue 5, Pages 1272-1281 (May 2009) DOI: 10.1016/j.jvs.2008.11.071 Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 1 Effect of sarpogrelate on development of intimal hyperplasia in autologous vein grafts at 4 weeks after operation. A, Microscopic findings in the middle portion of a vein graft from the control group (left panel) and the sarpogrelate-treated group (right panel). Arrowheads indicate the internal elastic lamina (elastic van Gieson staining). B, Quantitative analysis of the intimal hyperplasia of vein grafts. The intimal thickness was significantly smaller in the sarpogrelate-treated group (n = 5) than the control group (n = 5). C, The intima/media index was also significantly smaller in the sarpogrelate-treated group (n = 5) than in the control group (n = 5). Results are expressed as mean ± standard error of the mean (SEM), *P < .05 vs control group. Journal of Vascular Surgery 2009 49, 1272-1281DOI: (10.1016/j.jvs.2008.11.071) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 2 Effects of sarpogrelate on proliferative activity in neointima of autologous vein grafts. The proliferative cell antigen (PCNA) index was measured in vein grafts at 2 weeks after implantation. Counterstaining was done lightly with haematoxylin. A, Microscopic findings with PCNA staining in the middle portion of a vein graft from the control group (left panel) and the sarpogrelate-treated group (right panel). Arrowheads indicate PCNA-positive cells in the neointima (original magnification ×400). B, Quantitative analysis of the frequency of PCNA-positive cells. The PCNA index (the number of PCNA-positive cells divided by the number of total cells) was significantly suppressed in the arpogrelate-treated group (n = 8) compared with that in the control group (n = 8). Results are expressed as mean ± standard error of the mean (SEM), *P < .05 vs the control group. Journal of Vascular Surgery 2009 49, 1272-1281DOI: (10.1016/j.jvs.2008.11.071) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 3 Immunohistochemistry using antibodies against SM1 (A), SMemb (B) in neointima of autologous vein grafts. A, Most cells in the neointima were positive for monoclonal antibody against SM1, and SM1 index was similar between the control and the sarpogrelate-treated group (n = 8 in each case). B, SMemb index was significantly smaller in the sarpogrelate-treated group (n = 8) than in the control group (n = 8). Results are expressed as mean ± standard error of the mean (SEM), *P < .05 vs the control group. Journal of Vascular Surgery 2009 49, 1272-1281DOI: (10.1016/j.jvs.2008.11.071) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 4 Effects of sarpogrelate on prostaglandin F (PGF)2á-induced contraction in autologous vein grafts. PGF2á (5 ìM) induced a contraction in both control and sarpogrelate-treated group. In the sarpogrelate-treated group, L-NNA significantly enhanced the PGF2á-induced contraction in endothelium-intact strips, but not in endothelium-denuded strips. Such an effect of L-NNA was not observed in endothelium-intact strips from the control group. E(+), endothelium-intact strips; E(−), endothelium-denuded strips; L-NNA(−), in the absence of L-NNA; L-NNA(+), in the presence of L-NNA. Results are expressed as mean ± standard error of the mean (SEM), *P < .05 vs L-NNA(−). Journal of Vascular Surgery 2009 49, 1272-1281DOI: (10.1016/j.jvs.2008.11.071) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 5 Effects of sarpogrelate on acetylcholine (ACh)-induced mechanical response in autologous vein grafts. A, Actual tracings of the effects of ACh in endothelium-intact strips from the sarpogrelate-treated group in the absence (left panel) and presence of L-NNA (right panel). ACh (1 ìM) were applied during a contraction induced by prostaglandin F (PGF)2á (5 ìM). B, Summary of the effect of sarpogrelate on ACh-induced mechanical response. The amplitude of tonic contraction induced by PGF2α before application of ACh was normalized as the relative tension of 1.0 for each strip. In sarpogrelate-treated group, ACh induced a relaxation in the endothelium-intact strips but not in endothelium-denuded strips. In endothelium-intact strips of the sarpogrelate-treated group, L-NNA increased the PGF2α-induced contraction and blocked the ACh-induced relaxation. L-NNA(−), in the absence of L-NNA; L-NNA(+), in the presence of L-NNA. Results are expressed as mean ± standard error of the mean (SEM), *P < .05 vs before application of ACh, ††P < .01 vs L-NNA(−). Journal of Vascular Surgery 2009 49, 1272-1281DOI: (10.1016/j.jvs.2008.11.071) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 6 Effects of sarpogrelate on expression of eNOS protein in autologous vein grafts. A, Immunofluorescence against eNOS protein can be identified only in the endothelial region in both the groups. B, Summary of the effect of sarpogrelate on eNOS protein expression. There was no difference in the levels of eNOS protein expression between the two groups. Mean of data from four different sections (from four animals) with standard error of the mean (SEM). Journal of Vascular Surgery 2009 49, 1272-1281DOI: (10.1016/j.jvs.2008.11.071) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 7 Effects of sarpogrelate on superoxide production in vascular wall of vein grafts. A, Superoxide production (estimated from ethidium fluorescence) can be seen in both the endothelial and the medial region. L-NAME(+): in the presence of L-NAME; L-NAME(−): in the absence of L-NAME. B, Summary of the effect of sarpogrelate on superoxide production. Superoxide production in the endothelial region is significantly smaller in the sarpogrelate-treated group than in the control group. L-NAME significantly inhibits fluorescence intensities in the endothelial region in both the groups. Mean of data from four different sections (from four animals) with standard error of the mean (SEM), **P < .01 vs in the absence of L-NAME, †P < .05 vs control. Journal of Vascular Surgery 2009 49, 1272-1281DOI: (10.1016/j.jvs.2008.11.071) Copyright © 2009 Society for Vascular Surgery Terms and Conditions