A Lejay1, P Choquet2, F Thaveau3, F Singh4, A Schlagowski4, A-L Charles4, G Laverny5, D Metzger5, J Zoll4, N Chakfe6, B Geny4. 1. Equipe d'Accueil 3072, Mitochondrie, Stress Oxydant et Protection Musculaire, Université de Strasbourg, Fédération de Médecine Translationelle, Institut de Physiologie, 67000, Cedex, France; Department of Vascular Surgery and Kidney Transplantation, University Hospital, B.P. 426, 67091, Strasbourg, France. Electronic address: anne-catherine.lejay@orange.fr. 2. Department of Biophysics and Nuclear Medicine, University Hospital, B.P. 426, 67091, Strasbourg Cedex, France. 3. Equipe d'Accueil 3072, Mitochondrie, Stress Oxydant et Protection Musculaire, Université de Strasbourg, Fédération de Médecine Translationelle, Institut de Physiologie, 67000, Cedex, France; Department of Vascular Surgery and Kidney Transplantation, University Hospital, B.P. 426, 67091, Strasbourg, France. 4. Equipe d'Accueil 3072, Mitochondrie, Stress Oxydant et Protection Musculaire, Université de Strasbourg, Fédération de Médecine Translationelle, Institut de Physiologie, 67000, Cedex, France; Department of Physiology and Functional Explorations, University Hospital, B.P. 426, 67091, Strasbourg Cedex, France. 5. Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), CNRS UMR7104/INSERM U964, Collège de France, Université de Strasbourg, Illkirch Cedex, France. 6. Department of Vascular Surgery and Kidney Transplantation, University Hospital, B.P. 426, 67091, Strasbourg, France.
Abstract
OBJECTIVE: To establish a chronic mouse model of critical limb ischemia (CLI) with in vivo and ex vivo validation, closely mimicking human pathology. METHODS: Swiss mice (n = 28) were submitted to sequential unilateral femoral (day 0) and iliac (day 4) ligatures. Ischemia was confirmed by clinical scores (tissue and functional damages) and methoxyisobutylisonitrile (MIBI) scintigraphies at days 0, 4, 6, 10, 20, and 30. At days 10, 20, and 30, muscle mitochondrial respiration, calcium retention capacity (CRC), and production of reactive oxygen species (ROS) were investigated, together with transcripts of mitochondrial biogenesis and antioxidant enzymes. Histological analysis was also performed. RESULTS: Clinical and functional damage confirmed CLI. MIBI scintigraphies showed hypoperfusion of the ischemic limb, which remained stable until day 30. Mitochondrial respiration was impaired in ischemic muscles compared with controls (Vmax = 7.93 ± 0.99 vs. 10.09 ± 2.87 mmol/L O2/minute/mg dry weight [dw]; p = .01), together with impaired CRC (7.4 ± 1.6 mmol/L minute/mg dw vs. 11.9 ± 0.9 mmol/L minute/mg dw; p < .001) and biogenesis (41% decrease in peroxisome proliferator-activated receptor gamma coactivator [PGC]-1α, 49% decrease in PGC-1β, and 41% decrease in nuclear respiratory factor-1). Ischemic muscles also demonstrated increased production of ROS under electron paramagnetic resonance (0.084 ± 0.029 vs. 0.051 ± 0.031 mmol/L minute/mg dw; p = .03) and with dihydroethidium staining (3622 ± 604 arbitrary units of fluorescence vs. 1224 ± 324; p < .01), decreased antioxidant enzymes (32% decrease in superoxide dismutase [SOD]1, 41% decrease in SOD2, and 49% decrease in catalase), and myopathic features (wider range in fiber size, rounded shape, centrally located nuclei, and smaller cross-sectional areas). All defects were stable over time. CONCLUSION: Sequential femoral and iliac ligatures closely mimic human functional, clinical, scintigraphic, and skeletal muscle mitochondrial characteristics, and could prove useful for testing therapeutic approaches.
OBJECTIVE: To establish a chronic mouse model of critical limb ischemia (CLI) with in vivo and ex vivo validation, closely mimicking human pathology. METHODS: Swiss mice (n = 28) were submitted to sequential unilateral femoral (day 0) and iliac (day 4) ligatures. Ischemia was confirmed by clinical scores (tissue and functional damages) and methoxyisobutylisonitrile (MIBI) scintigraphies at days 0, 4, 6, 10, 20, and 30. At days 10, 20, and 30, muscle mitochondrial respiration, calcium retention capacity (CRC), and production of reactive oxygen species (ROS) were investigated, together with transcripts of mitochondrial biogenesis and antioxidant enzymes. Histological analysis was also performed. RESULTS: Clinical and functional damage confirmed CLI. MIBI scintigraphies showed hypoperfusion of the ischemic limb, which remained stable until day 30. Mitochondrial respiration was impaired in ischemic muscles compared with controls (Vmax = 7.93 ± 0.99 vs. 10.09 ± 2.87 mmol/L O2/minute/mg dry weight [dw]; p = .01), together with impaired CRC (7.4 ± 1.6 mmol/L minute/mg dw vs. 11.9 ± 0.9 mmol/L minute/mg dw; p < .001) and biogenesis (41% decrease in peroxisome proliferator-activated receptor gamma coactivator [PGC]-1α, 49% decrease in PGC-1β, and 41% decrease in nuclear respiratory factor-1). Ischemic muscles also demonstrated increased production of ROS under electron paramagnetic resonance (0.084 ± 0.029 vs. 0.051 ± 0.031 mmol/L minute/mg dw; p = .03) and with dihydroethidium staining (3622 ± 604 arbitrary units of fluorescence vs. 1224 ± 324; p < .01), decreased antioxidant enzymes (32% decrease in superoxide dismutase [SOD]1, 41% decrease in SOD2, and 49% decrease in catalase), and myopathic features (wider range in fiber size, rounded shape, centrally located nuclei, and smaller cross-sectional areas). All defects were stable over time. CONCLUSION: Sequential femoral and iliac ligatures closely mimic human functional, clinical, scintigraphic, and skeletal muscle mitochondrial characteristics, and could prove useful for testing therapeutic approaches.
Authors: Michele Diana; Eric Noll; Anne-Laure Charles; Pierre Diemunsch; Bernard Geny; Yu-Yin Liu; Francesco Marchegiani; Luigi Schiraldi; Vincent Agnus; Veronique Lindner; Lee Swanström; Bernard Dallemagne; Jacques Marescaux Journal: Surg Endosc Date: 2016-06-20 Impact factor: 4.584
Authors: Terence E Ryan; Cameron A Schmidt; Tom D Green; David A Brown; P Darrell Neufer; Joseph M McClung Journal: Front Physiol Date: 2015-11-18 Impact factor: 4.566
Authors: Sebastian Steven; Andreas Daiber; Jörn F Dopheide; Thomas Münzel; Christine Espinola-Klein Journal: Redox Biol Date: 2017-04-13 Impact factor: 11.799