Overexpression of endothelial nitric oxide synthase increases skeletal muscle blood flow and oxygenation in severe rat hind limb ischemia Lucy S Brevetti,

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Presentation transcript:

Overexpression of endothelial nitric oxide synthase increases skeletal muscle blood flow and oxygenation in severe rat hind limb ischemia Lucy S Brevetti, MD, David S Chang, MD, Gale L Tang, MD, Rajabrata Sarkar, MD, PhD, Louis M Messina, MD Journal of Vascular Surgery Volume 38, Issue 4, Pages 820-826 (October 2003) DOI: 10.1016/S0741-5214(03)00555-X

Fig 1 Effect of endothelial nitric oxide synthase (eNOS) overexpression on muscle blood flow and oxygen tension (Po2). A, Skeletal muscle blood flow determined by microspheres 14 days after gene transfer. Significant increase in encoding eNOS (AdeNOS)–treated rats. *P < .05 vs other groups. B, Muscle Po2 (in mm Hg) of normal and ischemic gastrocnemius muscles 14 and 30 days after gene transfer with rats breathing room air. Animals received the indicated titers of AdeNOS or control solutions as indicated. Note dose-dependent increase in Po2 14 days after gene transfer with AdeNOS treatment, which persisted at 30 days. C, Rats breathing 100% oxygen to amplify perfusion differences between normal and ischemic muscles. Statistically significant decrease in Po2 in phosphate-buffered saline solution (PBS)–treated and AdE1-treated ischemic animals; dose-dependent correction with AdeNOS treatment. *P < .05 vs PBS; **P < .05 vs AdE1; #P < .05 vs normal. Journal of Vascular Surgery 2003 38, 820-826DOI: (10.1016/S0741-5214(03)00555-X)

Fig 2 Transgene expression of endothelial nitric oxide synthase (eNOS) after adenoviral-mediated gene transfer in ischemic hind limbs. A, Western blot for eNOS protein on gastrocnemius muscle homogenates 3 days after gene transfer of either AdE1 (control vector) or adenoviral vector encoding eNOS (AdeNOS); graph of densitometric analysis with Western blot (n = 3 animals each). B, Immunohistochemistry of cross-sectioned gastrocnemius muscle with anti-eNOS antibody 3 days after gene transfer of either AdE1 or AdeNOS (×80). C, Calcium-dependent NOS activity of muscle homogenates at 3, 14, and 30 days after gene transfer. *P < .05 vs phosphate-buffered saline solution (PBS); **P < .05 vs AdE1. Journal of Vascular Surgery 2003 38, 820-826DOI: (10.1016/S0741-5214(03)00555-X)

Fig 3 Capillary–muscle fiber ratio and angiography after endothelial nitric oxide (eNOS) overexpression. A, Trend toward increased capillary–muscle fiber ratio 30 days after gene transfer (P = .08). B, More large collateral arteries were observed in adenoviral eNOS (AdeNOS)–treated rats 30 days after gene transfer. Fewer large collateral arteries were observed in phosphate-buffered saline solution (PBS)–treated rats. C, Significantly higher angioscores in AdeNOS-treated rats (n = 7) vs PBS-treated rats (n = 6). *P < .001). Journal of Vascular Surgery 2003 38, 820-826DOI: (10.1016/S0741-5214(03)00555-X)