title: Chronically ischemic mouse skeletal muscle exhibits myopathy in association with mitochondrial dysfunction and oxidative damage.
abstract: A myopathy characterized by mitochondrial pathology and oxidative stress is present in patients with peripheral arterial disease (PAD). Patients with PAD differ in disease severity, mode of presentation, and presence of comorbid conditions. In this study, we used a mouse model of hindlimb ischemia to isolate and directly investigate the effects of chronic inflow arterial occlusion on skeletal muscle microanatomy, mitochondrial function and expression, and oxidative stress. Hindlimb ischemia was induced by staged ligation/division of the common femoral and iliac arteries in C57BL/6 mice, and muscles were harvested 12 wk later. Muscle microanatomy was examined by bright-field microscopy, and mitochondrial content was determined as citrate synthase activity in muscle homogenates and ATP synthase expression by fluorescence microscopy. Electron transport chain (ETC) complexes I through IV were analyzed individually by respirometry. Oxidative stress was assessed as total protein carbonyls and 4-hydroxy-2-nonenal (HNE) adducts and altered expression and activity of manganese superoxide dismutase (MnSOD). Ischemic muscle exhibited histological features of myopathy and increased mitochondrial content compared with control muscle. Complex-dependent respiration was significantly reduced for ETC complexes I, III, and IV in ischemic muscle. Protein carbonyls, HNE adducts, and MnSOD expression were significantly increased in ischemic muscle. MnSOD activity was not significantly changed, suggesting MnSOD inactivation. Using a mouse model, we have demonstrated for the first time that inflow arterial occlusion alone, i.e., in the absence of other comorbid conditions, causes myopathy with mitochondrial dysfunction and increased oxidative stress, recapitulating the muscle pathology of PAD patients.
authors.affiliation: Department of Surgery, University of Nebraska Medical Center, Omaha, Nebraska 68198-3280, USA. ipipinos@unmc.edu
authors.forename: Iraklis I
authors.initials: II
authors.lastname: Pipinos
authors.forename: Stanley A
authors.initials: SA
authors.lastname: Swanson
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhu
authors.forename: Aikaterini A
authors.initials: AA
authors.lastname: Nella
authors.forename: Dustin J
authors.initials: DJ
authors.lastname: Weiss
authors.forename: Tanuja L
authors.initials: TL
authors.lastname: Gutti
authors.forename: Rodney D
authors.initials: RD
authors.lastname: McComb
authors.forename: B Timothy
authors.initials: BT
authors.lastname: Baxter
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Lynch
authors.forename: George P
authors.initials: GP
authors.lastname: Casale
mesh_terms: D025181:ATP Synthetase Complexes / Q000378:metabolism; D000447:Aldehydes / Q000378:metabolism; D000818:Animals; D002908:Chronic Disease; D002950:Citrate (si)-Synthase / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D006614:Hindlimb / Q000098:blood supply; D006801:Humans; D007511:Ischemia / Q000150:complications*; D051379:Mice; D008810:Mice, Inbred C57BL; D008931:Mitochondria, Muscle / Q000201:enzymology / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000473:pathology*; D009135:Muscular Diseases / Q000150:complications*; D018384:Oxidative Stress / Q000502:physiology*; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: D000447:Aldehydes; D013482:Superoxide Dismutase; D002950:Citrate (si)-Synthase; D025181:ATP Synthetase Complexes; C027576:4-hydroxy-2-nonenal