Research Article|Articles in Press

Diet-induced obesity augments ischemic myopathy and functional decline in a murine model of peripheral artery disease


      Peripheral artery disease (PAD) causes an ischemic myopathy contributing to patient disability and mortality. Most preclinical models to date use young, healthy rodents with limited translatability to human disease. Although PAD incidence increases with age, and obesity is a common comorbidity, the pathophysiologic association between these risk factors and PAD myopathy is unknown. Using our murine model of PAD, we sought to elucidate the combined effect of age, diet-induced obesity and chronic hindlimb ischemia (HLI) on (1) mobility, (2) muscle contractility, and markers of muscle (3) mitochondrial content and function, (4) oxidative stress and inflammation, (5) proteolysis, and (6) cytoskeletal damage and fibrosis. Following 16-weeks of high-fat, high-sucrose, or low-fat, low-sucrose feeding, HLI was induced in 18-month-old C57BL/6J mice via the surgical ligation of the left femoral artery at 2 locations. Animals were euthanized 4-weeks postligation. Results indicate mice with and without obesity shared certain myopathic changes in response to chronic HLI, including impaired muscle contractility, altered mitochondrial electron transport chain complex content and function, and compromised antioxidant defense mechanisms. However, the extent of mitochondrial dysfunction and oxidative stress was significantly greater in obese ischemic muscle compared to nonobese ischemic muscle. Moreover, functional impediments, such as delayed post-surgical recovery of limb function and reduced 6-minute walking distance, as well as accelerated intramuscular protein breakdown, inflammation, cytoskeletal damage, and fibrosis were only evident in mice with obesity. As these features are consistent with human PAD myopathy, our model could be a valuable tool to test new therapeutics.



      ARF (animal research facility), Atg7 (autophagy gene 7), Atg10 (autophagy gene 10), Atg12 (autophagy gene12), CI.2 (mitochondrial Complex I, state 2 respiration), CI.3 (mitochondrial Complex I, state 3 respiration), CI+II (combined Complex I and II respiration), CIII (mitochondrial Complex III), CIV (mitochondrial Complex IV), CV (mitochondrial Complex V), CLI (critical limb ischemia), CS (citrate synthase), CTGF (connective tissue growth factor), ECM (extracellular matrix), ETC (electron transport chain), FADH2 (1,5-dihydroflavin adenine dinucleotide), Fbxo32 (F-box O protein 32), FoxO1 (forkhead box O1), FoxO3 (forkhead box O3), GA (gastrocnemius), HFS (high-fat, high-sucrose), HLI (hindlimb ischemia), IACUC (Institutional Animal Care and Use Committee), IC (Intermittent claudication), Isch (ischemic), JH2O2 (mitochondrial hydrogen peroxide emission), JO2 (mitochondrial oxygen consumption), LC3 (microtubule-associated protein 1 light chain 3), LC3-II:LC3-I (microtubule-associated protein 1 light chain 3 isoform two-to-one ratio), LFS (low-fat, low-sucrose), mtDNA (mitochondrial DNA), NADH (nicotinamide adenine dinucleotide dehydrogenase), NI (non-ischemic), NOX (nicotinamide adenine dinucleotide phosphate oxidase), p62 (sequestosome 1), PAD (Peripheral artery disease), PARP-1 (Poly (ADP-ribose) polymerase 1), PCSA (physiological cross-sectional area), PSMA7 (proteasome alpha 7 subunit), PSMB5 (proteasome beta 5 subunit), ROS (reactive oxygen species), RyR1 (Ryanodine receptor 1), SERCA (sarcoplasmic reticulum), SOD1 (superoxide dismutase 1), TLR9 (Toll-like receptor 9), TRIM63 (tripartite motif containing 63), UPS (ubiquitin 26S proteasome)
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