Abstract
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.
Keywords
Abbreviations:
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)To read this article in full you will need to make a payment
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References
- New directions in therapeutic angiogenesis and arteriogenesis in peripheral arterial disease.Circ Res. 2021; 128: 1944-1957
- Global, regional, and national prevalence and risk factors for peripheral artery disease in 2015: an updated systematic review and analysis.Lancet Glob Health. 2019; 7: e1020-e1030
- Microvascular disease increases amputation in patients with peripheral artery disease.Arterioscler Thromb Vasc Biol. 2020; 40: 534-540
- The nitric oxide system in peripheral artery disease: connection with oxidative stress and biopterins.Antioxidants (Basel). 2020; 9
- Chronology of mitochondrial and cellular events during skeletal muscle ischemia-reperfusion.Am J Physiol Cell Physiol. 2016; 310: C968-C982
- Contemporary trends in hospital admissions and outcomes in patients with critical limb ischemia: an analysis from the National Inpatient Sample Database.Circ Cardiovasc Qual Outcomes. 2021; 14e007539
- Oxidative stress and antioxidant treatment in patients with peripheral artery disease.Physiol Rep. 2018; 6: e13650
- The myopathy of peripheral arterial occlusive disease: part 1. Functional and histomorphological changes and evidence for mitochondrial dysfunction.Vasc Endovascular Surg. 2007; 41: 481-489
- Experimental peripheral arterial disease: new insights into muscle glucose uptake, macrophage, and T-cell polarization during early and late stages.Physiol Rep. 2014; 2: e00234
- Why to evaluate the functional capacity in PAD patients?.Vasa. 2020; 49: 275-280
- Mitochondrial regulation of the muscle microenvironment in critical limb ischemia.Front Physiol. 2015; 6: 336
- Oxidative damage and myofiber degeneration in the gastrocnemius of patients with peripheral arterial disease.J Transl Med. 2013; 11: 230
- Oxidative damage in the gastrocnemius of patients with peripheral artery disease is myofiber type selective.Redox Biol. 2014; 2: 921-928
- The myopathy of peripheral arterial occlusive disease: part 2. Oxidative stress, neuropathy, and shift in muscle fiber type.Vasc Endovascular Surg. 2008; 42: 101-112
- Chronically ischemic mouse skeletal muscle exhibits myopathy in association with mitochondrial dysfunction and oxidative damage.Am J Physiol Regul, Integr Comp Physiol. 2008; 295: R290-R296
- Extensive skeletal muscle cell mitochondriopathy distinguishes critical limb ischemia patients from claudicants.JCI Insight. 2018; 3
- Peripheral artery disease affects the function of the legs of claudicating patients in a diffuse manner irrespective of the segment of the arterial tree primarily involved.PLoS One. 2022; 17e0264598
- Protein concentration and mitochondrial content in the gastrocnemius predicts mortality rates in patients with peripheral arterial disease.Ann Surg. 2015; 261: 605-610
- Sarcopenia and peripheral arterial disease: a systematic review.J Cachexia Sarcopenia Muscle. 2020; 11: 866-886
- Aging exacerbates ischemia-reperfusion-induced mitochondrial respiration impairment in skeletal muscle.Antioxidants (Basel). 2019; 8
- Morphometric analysis of gastrocnemius muscle biopsies from patients with peripheral arterial disease: objective grading of muscle degeneration.Am J Physiol Regul Integr Comp Physiol. 2013; 305: R291-R299
- Abnormal mitochondrial respiration in skeletal muscle in patients with peripheral arterial disease.J Vasc Surg. 2003; 38: 827-832
- Skeletal muscle MiR-210 expression is associated with mitochondrial function in peripheral artery disease patients.Transl Res. 2022; 246: 66-77
- Cytokine signature of inflammation mediated by autoreactive Th-cells, in calf muscle of claudicating patients with Fontaine stage II peripheral artery disease.Transl Res. 2021; 228: 94-108
- Skeletal muscle metabolism as a target for drug therapy in peripheral arterial disease.Vasc Med. 1996; 1: 55-59
- Acquired skeletal muscle metabolic myopathy in atherosclerotic peripheral arterial disease.Vasc Med. 2000; 5: 55-59
- Evaluation of the clinical relevance and limitations of current pre-clinical models of peripheral artery disease.Clin Sci (Lond). 2016; 130: 127-150
- Methods for acute and subacute murine hindlimb ischemia.J Vis Exp. 2016;
- Variations in surgical procedures for inducing hind limb ischemia in mice and the impact of these variations on neovascularization assessment.Int J Mol Sci. 2019; 20
- Associations of obesity with incident hospitalization related to peripheral artery disease and critical limb ischemia in the ARIC study.J Am Heart Assoc. 2018; 7e008644
- Age-dependent impairment of angiogenesis.Circulation. 1999; 99: 111-120
- Limb ischemia after iliac ligation in aged mice stimulates angiogenesis without arteriogenesis.J Vasc Surg. 2009; 49: 464-473
- Preclinical models of human peripheral arterial occlusive disease: implications for investigation of therapeutic agents.J Appl Physiol (1985). 2004; 97: 773-780
- Body mass index and peripheral artery disease.Atherosclerosis. 2020; 292: 31-36
- Obesity and peripheral arterial disease: a Mendelian randomization analysis.Atherosclerosis. 2016; 247: 218-224
- Higher body mass index is associated with more adverse changes in calf muscle characteristics in peripheral arterial disease.J Vasc Surg. 2012; 55: 1015-1024
- Obesity in the elderly: an emerging health issue.Int J Obes (Lond). 2012; 36: 1151-1152
- Guide for the care and use of laboratory animals.8th ed. National Academies Press, Washington (DC)2011
- A mouse model of diet-induced obesity and insulin resistance.Methods Mol Biol. 2012; 821: 421-433
- Obesity-induced alterations to the immunoproteasome: a potential link to intramuscular lipotoxicity.Appl Physiol Nutr Metab. 2021; 46: 485-493
- Impaired proteostasis in obese skeletal muscle relates to altered immunoproteasome activity.Appl Physiol Nutr Metab. 2022; 47: 555-564
- Defining body-weight reduction as a humane endpoint: a critical appraisal.Lab Anim. 2020; 54: 99-110
- Murine model of hindlimb ischemia.J Vis Exp. 2009;
- Spinal cord compression studies. III. Time limits for recovery after gradual compression in dogs.AMA Arch Neurol Psychiatry. 1954; 71: 588-597
- Effects of quercetin on exercise performance, physical activity and blood supply in a novel model of sustained hind-limb ischaemia.BJS Open. 2021; 5
- Development of a two-stage limb ischemia model to better simulate human peripheral artery disease.Sci Rep. 2020; 10: 3449
- Use of the Open Field Maze to measure locomotor and anxiety-like behavior in mice.J Vis Exp. 2015; : e52434
- A simple and low-cost assay for measuring ambulation in mouse models of muscular dystrophy.J Vis Exp. 2017;
- Preparation of acute hippocampal slices from rats and transgenic mice for the study of synaptic alterations during aging and amyloid pathology.J Vis Exp. 2011;
- Further development of a tissue engineered muscle repair construct in vitro for enhanced functional recovery following implantation in vivo in a murine model of volumetric muscle loss injury.Tissue Eng Part A. 2012; 18: 1213-1228
- Isometric skeletal muscle contractile properties in common strains of male laboratory mice.Front Physiol. 2022; 13937132
- Density and composition of mammalian muscle.Metabol, Clin Exp. 1960; 9: 184-188
- Contractile properties of skeletal muscles from young, adult and aged mice.J Physiol. 1988; 404: 71-82
- Mitochondrial functional impairment with aging is exaggerated in isolated mitochondria compared to permeabilized myofibers.Aging Cell. 2010; 9: 1032-1046
- Targeted expression of catalase to mitochondria protects against ischemic myopathy in high-fat diet-fed mice.Diabetes. 2016; 65: 2553-2568
- Comparison of mitochondrial incubation media for measurement of respiration and hydrogen peroxide production.Methods Mol Biol. 2018; 1782: 137-155
- Experimental oxygen concentration influences rates of mitochondrial hydrogen peroxide release from cardiac and skeletal muscle preparations.Am J Physiol Regul Integr Compar Physiol. 2020; 318: R972-R980
- Simultaneous high-resolution measurement of mitochondrial respiration and hydrogen peroxide production.Methods Mol Biol. 2015; 1264: 245-261
- Microplate assays for spectrophotometric measurement of mitochondrial enzyme activity.Methods Mol Biol. 2019; 1978: 355-368
- Denervation-induced activation of the standard proteasome and immunoproteasome.PLoS One. 2016; 11e0166831
- Altered proteasome structure, function, and oxidation in aged muscle.FASEB J. 2005; 19: 644-646
- Mitochondriopathy of peripheral arterial disease.Vascular. 2007; 15: 336-343
- Mitochondrial defects and oxidative damage in patients with peripheral arterial disease.Free Radic Biol Med. 2006; 41: 262-269
- Protein carbonylation as a major hallmark of oxidative damage: update of analytical strategies.Mass Spectrom Rev. 2014; 33: 79-97
- Redox control of skeletal muscle atrophy.Free Radic Biol Med. 2016; 98: 208-217
- Inflammation induced loss of skeletal muscle.Bone. 2015; 80: 131-142
- Role of inflammation in muscle homeostasis and myogenesis.Mediators Inflamm. 2015; 2015805172
- Activation of caspase-3 is an initial step triggering accelerated muscle proteolysis in catabolic conditions.J Clin Invest. 2004; 113: 115-123
- Activation and caspase-mediated inhibition of PARP: a molecular switch between fibroblast necrosis and apoptosis in death receptor signaling.Mol Biol Cell. 2002; 13: 978-988
- FoxO transcription factors: their roles in the maintenance of skeletal muscle homeostasis.Cell Mol Life Sci. 2014; 71: 1657-1671
- MuRF1/TRIM63, master regulator of muscle mass.Int J Mol Sci. 2020; 21
- Regulation of autophagy and the ubiquitin-proteasome system by the FoxO transcriptional network during muscle atrophy.Nat Commun. 2015; 6: 6670
- The role of autophagy in skeletal muscle diseases.Front Physiol. 2021; 12638983
- FoxO1 is a novel regulator of 20S proteasome subunits expression and activity.Front Cell Dev Biol. 2021; 9625715
- Defining and measuring autophagosome flux-concept and reality.Autophagy. 2014; 10: 2087-2096
- The rise of mitochondria in peripheral arterial disease physiopathology: experimental and clinical data.J Clin Med. 2019; 8
- Mitochondria: mitochondrial participation in ischemia-reperfusion injury in skeletal muscle.Int J Biochem Cell Biol. 2014; 50: 101-105
- The role of mitochondrial function in peripheral arterial disease: insights from translational studies.Int J Mol Sci. 2021; 22
- Linking mitochondrial bioenergetics to insulin resistance via redox biology.Trends Endocrinol Metab. 2012; 23: 142-153
- Increased oxidative stress in obesity: implications for metabolic syndrome, diabetes, hypertension, dyslipidemia, atherosclerosis, and cancer.Obes Res Clin Pract. 2013; 7: e330-e341
- Reactive oxygen species and oxidative stress in obesity-recent findings and empirical approaches.Obesity (Silver Spring). 2016; 24: 2301-2310
- Reactive oxygen species and calcium signals in skeletal muscle: a crosstalk involved in both normal signaling and disease.Cell Calcium. 2016; 60: 172-179
- Protein carbonylation, mitochondrial dysfunction, and insulin resistance.Adv Nutr. 2013; 4: 157-163
- Mitochondrial DNA and TLR9 drive muscle inflammation upon opa1 deficiency.EMBO J. 2018; 37
- Mitochondrial DNA in inflammation and immunity.EMBO Rep. 2020; 21: e49799
- TLR9 in MAFLD and NASH: at the intersection of inflammation and metabolism.Front Endocrinol (Lausanne). 2020; 11613639
- Endogenously oxidized mitochondrial DNA induces in vivo and in vitro inflammatory responses.J Leukoc Biol. 2004; 75: 995-1000
- VDAC oligomers form mitochondrial pores to release mtDNA fragments and promote lupus-like disease.Science. 2019; 366: 1531-1536
- Mitochondria-derived superoxide links to tourniquet-induced apoptosis in mouse skeletal muscle.PLoS One. 2012; 7: e43410
- Skeletal muscle mitochondrial DNA injury in patients with unilateral peripheral arterial disease.Circulation. 1999; 99: 807-812
- Associations of peripheral artery disease with calf skeletal muscle mitochondrial DNA heteroplasmy.J Am Heart Assoc. 2020; 9e015197
- Peripheral artery disease, calf skeletal muscle mitochondrial DNA copy number, and functional performance.Vasc Med. 2018; 23: 340-348
- Bench-to-bedside review: Mitochondrial injury, oxidative stress and apoptosis–there is nothing more practical than a good theory.Crit Care. 2008; 12: 206
- Autophagy, apoptosis, and mitochondria: molecular integration and physiological relevance in skeletal muscle.Am J Physiol Cell Physiol. 2019; 317: C111-C130
- Altered Ca(2+) handling and oxidative stress underlie mitochondrial damage and skeletal muscle dysfunction in aging and disease.Metabolites. 2021; 11
- Reactive oxygen species and calcium signals in skeletal muscle: a crosstalk involved in both normal signaling and disease.Cell Calcium. 2016; 60: 172-179
- The role of calpains in skeletal muscle remodeling with exercise and inactivity-induced atrophy.Int J Sports Med. 2020; 41: 994-1008
- Skeletal muscle mitochondria and aging: a review.J Aging Res. 2012; 2012194821
- Ectopic fat and insulin resistance: pathophysiology and effect of diet and lifestyle interventions.Int J Endocrinol. 2012; 2012983814
- Lipotoxicity plays a key role in the development of both insulin resistance and muscle atrophy in patients with type 2 diabetes.Obes Rev. 2019; 20: 1205-1217
- High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C–dependent activation of NAD(P)H oxidase in cultured vascular cells.Diabetes. 2000; 49: 1939-1945
- Poly(ADP-ribose) polymerase-1 cleavage during apoptosis: an update.Apoptosis. 2002; 7: 321-328
- The 89-kDa PARP1 cleavage fragment serves as a cytoplasmic PAR carrier to induce AIF-mediated apoptosis.J Biol Chem. 2021; 296100046
- Role of poly(ADP-ribose) polymerase 1 (PARP-1) in cardiovascular diseases: the therapeutic potential of PARP inhibitors.Cardiovasc Drug Rev. 2007; 25: 235-260
- Associations of poly (ADP-Ribose) Polymerase1 abundance in calf skeletal muscle with walking performance in peripheral artery disease.Exp Gerontol. 2020; 140111048
- Mitophagy: an emerging role in aging and age-associated diseases.Front Cell Dev Biol. 2020; 8: 200
- Mitochondrial complex abundance, mitophagy proteins, and physical performance in people with and without peripheral artery disease.J Am Heart Assoc. 2023; 12e027088
- Regulating mitochondrial outer membrane proteins by ubiquitination and proteasomal degradation.Curr Opin Cell Biol. 2011; 23: 476-482
- Ubiquitin-dependent degradation of mitochondrial proteins regulates energy metabolism.Cell Rep. 2018; 23: 2852-2863
- Calpain-Mediated mitochondrial damage: an emerging mechanism contributing to cardiac disease.Cells. 2021; 10
- Inhibition of the ubiquitous calpains protects complex I activity and enables improved mitophagy in the heart following ischemia-reperfusion.Am J Physiol Cell Physiol. 2019; 317: C910-C921
- Mitochondrial Calpain-1 disrupts ATP synthase and induces superoxide generation in type 1 diabetic hearts: a novel mechanism contributing to diabetic cardiomyopathy.Diabetes. 2016; 65: 255-268
- Shared and distinct mechanisms of skeletal muscle atrophy: a narrative review.Ageing Res Rev. 2021; 71101463
- Abnormal accumulation of desmin in gastrocnemius myofibers of patients with peripheral artery disease: associations with altered myofiber morphology and density, mitochondrial dysfunction and impaired limb function.J Histochem Cytochem. 2015; 63: 256-269
- Endothelial cell-derived pro-fibrotic factors increase TGF-beta1 expression by smooth muscle cells in response to cycles of hypoxia-hyperoxia.Biochim Biophys Acta Mol Basis Dis. 2022; 1868166278
- Role of hypoxia in skeletal muscle fibrosis: synergism between hypoxia and TGF-beta signaling upregulates CCN2/CTGF expression specifically in muscle fibers.Matrix Biol. 2020; 87: 48-65
- Driving fibrosis in neuromuscular diseases: role and regulation of connective tissue growth factor (CCN2/CTGF).Matrix Biol Plus. 2021; 11100059
- Different responses of skeletal muscles to femoral artery ligation-induced ischemia identified in BABL/c and C57BL/6 mice.Front Physiol. 2022; 131014744
- Sex differences in peripheral artery disease.Circ Res. 2022; 130: 496-511
Article info
Publication history
Published online: May 21, 2023
Accepted:
May 15,
2023
Received in revised form:
May 10,
2023
Received:
April 17,
2023
Publication stage
In Press Journal Pre-ProofIdentification
Copyright
© 2023 Elsevier Inc. All rights reserved.
