Pioglitazone preserves vein graft integrity in a rat aortic interposition model Zhi Chen, MD, Tomomi Hasegawa, MD, PhD, Akiko Tanaka, MD, Yutaka Okita,

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Pioglitazone preserves vein graft integrity in a rat aortic interposition model Zhi Chen, MD, Tomomi Hasegawa, MD, PhD, Akiko Tanaka, MD, Yutaka Okita, MD, PhD, Kenji Okada, MD, PhD The Journal of Thoracic and Cardiovascular Surgery Volume 140, Issue 2, Pages 408-416.e1 (August 2010) DOI: 10.1016/j.jtcvs.2009.11.067 Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 Effects of pioglitazone on endothelial desquamation and oxidative stress in vein grafts at 24 hours. A, In vivo Evans blue staining of vein grafts. Anastomotic sites (arrows). Bar = 5 mm. B, In situ detection of superoxide generation (red fluorescence). Bar = 100 μm. C, Malondialdehyde levels as an indicator of lipid peroxidation. D, MPO activity. ΔAbs, change in absorbance. ∗P < .05. All data are expressed as mean ± standard error of the mean (SEM) for n = 6 per group. MDA, Malondialdehyde. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 408-416.e1DOI: (10.1016/j.jtcvs.2009.11.067) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Protease activity and mRNA expression in vein grafts at 7 days. A, mRNA expression of MMP-2, MMP-9, TIMP-1, and TIMP-2. B, Gelatinolytic activities of MMP-2 and MMP-9 by gelatin zymography, and their densitometric analysis. ∗P < .05. All data are expressed as mean ± SEM for n = 6 per group. Pio, Pioglitazone; NS, not significant; MMP, matrix metalloproteinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 408-416.e1DOI: (10.1016/j.jtcvs.2009.11.067) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 Graft flow velocity in vein grafts. A, Blood flow waveforms in proximal aorta, vein graft, and distal aorta at 6 months. B, Graft flow velocity ratio at 8 weeks and 6 months. ∗P < .05 by analysis of variance. All data are expressed as mean ± SEM for n = 6 per group. Ao, Aorta. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 408-416.e1DOI: (10.1016/j.jtcvs.2009.11.067) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 A, Graft dilation ratio at 8 weeks and 6 months. ∗P < .05. All data are expressed as mean ± SEM for n = 6 per group. B, Immunofluorescent staining of PPAR-γ in vein grafts at 8 weeks. Intracellular expression of PPAR-γ is identified as the green emission. Cell nuclei are identified by the blue emission. Bar = 200 μm. PPAR, Peroxisome proliferator-activated receptor. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 408-416.e1DOI: (10.1016/j.jtcvs.2009.11.067) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Figure 5 Histologic findings in vein grafts at 6 months. A, Elastin von Gieson staining and αSMA immunostaining. Bar = 500 μm. B, Area of neointimal hyperplasia and αSMA-positive area. C, Neointimal thickness at anastomosis site (hematoxylin–eosin staining). Bar = 200 μm. ∗P < .05. All data are expressed as mean ± SEM for n = 6 per group. EVG, Elastin von Gieson; SMA, smooth muscle actin. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 408-416.e1DOI: (10.1016/j.jtcvs.2009.11.067) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Schematic representation of the present study design Schematic representation of the present study design. Drug treatment (pioglitazone or vehicle) was provided to rats by gastric gavage once per day beginning 3 days before operation and ending 8 weeks after operation. The end points of the present study were 24 hours, 7 days, 8 weeks, and 6 months after operation. Op, Operation. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 408-416.e1DOI: (10.1016/j.jtcvs.2009.11.067) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions