Extracellular Superoxide Dismutase Accelerates Endothelial Recovery and Inhibits In-Stent Restenosis in Stented Atherosclerotic Watanabe Heritable Hyperlipidemic Rabbit Aorta
Jan Hinrich Bräsen, MD*, ,
Olli Leppänen, MD*, ,
Matias Inkala, MD*,
Tommi Heikura, MSc*,
Max Levin, MD, PhD ,
Fabian Ahrens, MSc||,
Juha Rutanen, MD, PhD*,
Hubertus Pietsch, DVM¶,
David Bergqvist, MD, PhD,
Anna-Liisa Levonen, MD, PhD*,
Samar Basu, PhD,
Thomas Zeller, MD, FESC#,
Günter Klöppel, MD,
Mikko O. Laukkanen, PhD** and
Seppo Ylä-Herttuala, MD, PhD, FESC*,,*
* A. I. Virtanen Institute, University of Kuopio, Kuopio, Finland
Institute for Pathology, Christian Albrechts Universität, Kiel, Germany
Uppsala University, Uppsala, Sweden
Wallenberg Laboratory, Gothenburg, Sweden
# Humboldt University, Berlin, Germany
¶ Schering AG, Berlin, Germany
** Herz-Zentrum, Bad Krozingen, Germany
** University of Turku, Turku, Finland
Gene Therapy Unit, Department of Medicine, University of Kuopio, Kuopio, Finland
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Figure 1 Quantification of In-Stent Restenosis
(A) Micro-computed tomographic imaging, methacrylate embedding (insert), and serial sections. (B) Quantification of vessel compartments. Arrow indicates medial dissection. Bar = 1 mm. (C) Strut overlying atherosclerotic lesion. (D) Histomorphometry. *p < 0.05. (E) Representative images of Mason trichrome-stained 28-day sections. Arrowheads indicate IEL. Bars = 100 µm; AdEC-SOD = adenovirus-encoding rabbit extracellular superoxide dismutase; AdLacZ = control adenovirus encoding nuclear targeted ß-galactosidase; IEL = ifernal elastic lamina; m = media; NS = not significant; s = strut.
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Figure 2 Re-Endothelialization, Macrophages, Transgene Expression, and Oxidant Production
(A) Endothelial coverage. (B) Neointimal macrophages. (C) LacZ transduced vessels and reverse transcriptase-polymerase chain reaction for extracellular superoxide dismutase (EC-SOD) mRNA. EC-SOD = 6-day EC-SOD–stented vessel; LacZ = 6-day control stented vessel. (D) DHE-stained nuclei in 6-day neointima and peroxynitrite (ONOO–)-modified epitopes. Representative images. Arrowheads = IEL. Bars = 100 µm. *p < 0.05. AdEC-SOD = adenovirus-encoding rabbit extracellular superoxide dismutase; Anti-ß-gal-mAB = immunohistochemical statin with monoclonal antibody against ß-galactosidase; DHE = dihydroethidium; HOCI = hypochlorite; LacZ = ß-galactosidase; mAb 2D 10G9 = monoclonal antibody against hypochlorite modified epitopes; mw = marker; Neg. = reaction product without test sample; PECAM-1 = platelet endothelial cell adhesion molecule-1; Pos. = extracellular superoxide dismutase pDNA; X-gal = 5-brom-4-chlor-3-indoxyl-ß-D-galactopyranoside chromogen substrate for ß-galactosidase; other abbreviations as in Figure 1.
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Figure 3 Systemic Oxidative Stress
Circulating nonesterified 8-iso-prostaglandin-F2 (8-iso-PGF2). *p < 0.05 versus immediate pre-intervention values within both groups. **p < 0.01 EC-SOD versus LacZ at 3 days. All other comparisons p = NS. Abbreviations as in Figure 2.
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