Remote and local ischemic preconditioning equivalently protects rat skeletal muscle mitochondrial function during experimental aortic cross-clamping

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Remote and local ischemic preconditioning equivalently protects rat skeletal muscle mitochondrial function during experimental aortic cross-clamping Ziad Mansour, MD, MS, Jamal Bouitbir, PhD, Anne Laure Charles, PhD, Samy Talha, MD, Michel Kindo, MD, Julien Pottecher, MD, Joffrey Zoll, PhD, Bernard Geny, MD, PhD Journal of Vascular Surgery Volume 55, Issue 2, Pages 497-505.e1 (February 2012) DOI: 10.1016/j.jvs.2011.07.084 Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 1 Experimental design. The control (C) animals (n = 10) underwent 6 hours of general anesthesia and a sham operation. The ischemia-reperfusion (IR) animals (n = 8) underwent 3 hours of ischemia induced by infrarenal aortic occlusion (dark bar), followed by 2 hours of reperfusion (open bar). The ischemic preconditioning (IPC) animals (n = 10) underwent the same IR protocol but prolonged ischemia was preceded by three 10-minute consecutive reperfusion sequences, each separated by 10 minutes of reocclusion. The IPC cycles were performed on the right hind limb. The right hind limb thus underwent local IPC and the left hind limb underwent remote IPC. Journal of Vascular Surgery 2012 55, 497-505.e1DOI: (10.1016/j.jvs.2011.07.084) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 2 Top, Schematic representation shows the mitochondrial respiratory chain with specific substrates and inhibitors. I, complex I (nicotinamide adenine dinucleotide reductase [NADH]-coenzyme Q [CoQ] reductase); II, complex II (succinate-CoQ reductase); III, complex III (CoQH2-c reductase); IV, complex IV (cytochrome-c oxidase, COX); V, ATP synthase; TMPD, N,N,N′,N′-tetramethyl-p-phenylenediamine dihydrochloride. Bottom, Schematic oxygraph trace shows oxygen consumption by the permeabilized skeletal myofibers, using indicated substrates and inhibitors: Vo, before adenosine diphosphate (ADP); Vmax, complexes I, III, IV activities, using glutamate and malate; Vsucc, complexes II, III, IV activities, using succinate; VTMPD, complex IV activity using TMPD/ascorbate. Journal of Vascular Surgery 2012 55, 497-505.e1DOI: (10.1016/j.jvs.2011.07.084) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 3 Assessment of the experimental model. A, Right and left hind limb Vmax, Vsucc, and VTMPD in control. B, Right and left hind limb Vmax, Vsucc, and VTMPD after ischemia-reperfusion. TMPD, N,N,N′,N′-tetramethyl-p-phenylenediamine dihydrochloride/ascorbate. Results are expressed as means ± standard error of the mean (error bars). Journal of Vascular Surgery 2012 55, 497-505.e1DOI: (10.1016/j.jvs.2011.07.084) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 4 Effects of ischemia-reperfusion (IR) and local ischemic preconditioning (IPC) and remote IPC (rIPC) on mitochondrial respiratory chain complexes activities. A, Vmax, complexes I, III, IV activities, using glutamate and malate. B, Vsucc, complexes II, III, IV activities, using succinate. C, VTMPD, complex IV activity using N,N,N′,N′-tetramethyl-p-phenylenediamine dihydrochloride/ascorbate (TMPD), as mitochondrial substrates. Data are presented in 3 experimental conditions: control (C), after IR, and after local IPC and rIPC. Results are expressed as means ± standard error of the mean (error bars). *P < .05. **P < .01. Journal of Vascular Surgery 2012 55, 497-505.e1DOI: (10.1016/j.jvs.2011.07.084) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 5 Effects of ischemia-reperfusion (IR), local ischemic preconditioning (IPC), and remote IPC (rIPC) on messenger RNA (mRNA) expression of genes involved in apoptosis. Data are presented in three experimental conditions: control (C), after IR, and after IPC and rIPC. A, Bax. mRNA expression level in muscles. B, The mRNA ratio of Bax/Bcl2 expression levels in muscles. Results are expressed as means ± standard error of the mean (error bars). *P < .05. **P < .01. Journal of Vascular Surgery 2012 55, 497-505.e1DOI: (10.1016/j.jvs.2011.07.084) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 6 Effects of ischemia-reperfusion (IR), local ischemic preconditioning (IPC), and remote IPC (rIPC) on antioxidant defense. Messenger RNA (mRNA) levels are shown for (A) superoxide dismutase 1 (SOD1) and (B) SOD2. Expression of genes involved in antioxidant defense: C, Glutathione (GPx). D, Reduced glutathione (GSH). Data are presented in three experimental conditions: control (C), after IR and after local IPC and rIPC. Results are expressed as means ± standard error of the mean (error bars). Journal of Vascular Surgery 2012 55, 497-505.e1DOI: (10.1016/j.jvs.2011.07.084) Copyright © 2012 Society for Vascular Surgery Terms and Conditions