Remote and local ischemic postconditioning further impaired skeletal muscle mitochondrial function after ischemia-reperfusion Ziad Mansour, MD, PhD,

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Remote and local ischemic postconditioning further impaired skeletal muscle mitochondrial function after ischemia-reperfusion Ziad Mansour, MD, PhD, Anne L. Charles, PhD, Jamal Bouitbir, PhD, Julien Pottecher, MD, Michel Kindo, MD, PhD, Jean-Philippe Mazzucotelli, MD, PhD, Joffrey Zoll, PhD, Bernard Geny, MD, PhD Journal of Vascular Surgery Volume 56, Issue 3, Pages 774-782.e1 (September 2012) DOI: 10.1016/j.jvs.2012.01.079 Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 1 Experimental design. Sham animals underwent 4 hours of general anesthesia and a sham operation. The ischemia-reperfusion (IR) animals underwent 2 hours of tourniquet-initiated ischemia in both hind limbs (dark bar), followed by 2 hours of reperfusion (open bar). The ischemic postconditioned (PoC) animals underwent the same IR protocol, but reperfusion on the right hind limb was preceded by four 30-second consecutive reperfusion sequences, each separated by 30 seconds of reocclusion. The right hind limb thus underwent local PoC and the left hind limb underwent remote PoC (rPOC). Journal of Vascular Surgery 2012 56, 774-782.e1DOI: (10.1016/j.jvs.2012.01.079) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 2 Maximal mitochondrial respiratory rate in sham, after ischemia-reperfusion (IR), and after local postconditioning (PoC) and remote ischemic PoC (rPoC). Maximal oxidative capacity (Vmax) was assessed using glutamate-malate as the mitochondrial substrate. Results are expressed as mean ± standard error of the mean (error bar). *P < .05. **P < .01. ***P < .001. Journal of Vascular Surgery 2012 56, 774-782.e1DOI: (10.1016/j.jvs.2012.01.079) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 3 Skeletal muscle production of reactive oxygen species (ROS) in sham, after ischemia-reperfusion (IR), and after local ischemic postconditioning (PoC) and remote PoC (rPoC). A, Fluorescence photographs of each subset. B, Intensity of ROS fluorescence. DHE, Dihydroethidium. Results are expressed as mean ± standard error of the mean (error bars). *P < .05. **P < .01.***P < .001. Journal of Vascular Surgery 2012 56, 774-782.e1DOI: (10.1016/j.jvs.2012.01.079) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 4 Effects of ischemia-reperfusion (IR) and local postconditioning (PoC) and remote ischemic postconditioning (rPoC) on the expression of genes involved in antioxidant defense. A, Superoxide dismutase 1 (SOD1). B, Superoxide dismutase 2 (SOD2). C, Glutathione peroxidase (GPx). Results are expressed as mean ± standard error of the mean (error bars). *P < .05. **P < .01. ***P < .001. Journal of Vascular Surgery 2012 56, 774-782.e1DOI: (10.1016/j.jvs.2012.01.079) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 5 Effects of ischemia-reperfusion (IR) and local postconditioning (PoC) and remote ischemic PoC (rPoC) on the expression of genes involved in apoptosis. A, Bax. B, BclII. C, Messenger RNA ratio of Bax/BclII. Results are expressed as mean ± standard error of the mean (error bars). *P < .05. **P < .01. ***P < .001. Journal of Vascular Surgery 2012 56, 774-782.e1DOI: (10.1016/j.jvs.2012.01.079) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 6 Effects of ischemia-reperfusion (IR) and local postconditioning (PoC) and remote ischemic PoC (rPoC) on inflammation. A, Expression of interleukin-6 (IL-6). Results are expressed as mean ± standard error of the mean (error bars). B, Immunohistochemical demonstration of the distribution of macrophages, which are indicated by the black arrows (original magnification, ×400). Journal of Vascular Surgery 2012 56, 774-782.e1DOI: (10.1016/j.jvs.2012.01.079) Copyright © 2012 Society for Vascular Surgery Terms and Conditions