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Doxorubicin chemotherapy affects intracellular and interstitial nitric oxide concentrations in skeletal muscle

Effect of doxorubicin on intracellular and interstitial NO in skeletal muscle

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Abstract

The role of nitric oxide (NO) in doxorubicin (DOX; cancer chemotherapeutic)-induced cardiotoxicity is well established. In skeletal muscle (SM), NO regulation plays a critical role in health, biogenesis, and function. Despite the increasing evidence that indicates the negative impact of DOX on SM function, the effect of DOX on NO production in SM has yet to be examined. The purpose of the current study was to simultaneously examine intracellular and interstitial NO concentrations in the SM following the administration of DOX. A single dose of 1.5 or 4.5 mg/kg was administered intraperitoneally to male Sprague Dawley rats, and interstitial (IS) and intracellular (IC) NO was quantified every 24 up to 192 h post-injection. There was no significant difference in IC NO following the injection of 1.5 mg/kg DOX when compared to the control; however, the administration of 4.5 mg/kg DOX resulted in lower (P < 0.05) concentrations of NO in the IC. Interestingly, a consistently higher (P < 0.05) concentration of NO in the IS was established following the administration of 1.5 mg/kg compared to the control while no significant changes in IS NO resulted from the administration of the 4.5 mg/kg dose. The fluctuation of IS and IC NO was not a result of substrate availability as arginine concentrations remained stable throughout the experiment. By utilizing the microdialysis technique, we have simultaneously quantified for the first time the IS and IC concentrations of NO in SM following DOX administration. These data provide important insight in the possible mechanisms leading to DOX-related SM dysfunction.

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Abbreviations

AKR:

Aldo-keto reductase

Arg:

L-arginine

CBR:

Carbonyl reductase

DOX:

Doxorubicin

eNOS:

Endothelial nitric oxide synthase

iNOS:

Inducible nitric oxide synthase

Jun-D:

Transcription factor Jun-D

MCK:

Muscle creatine kinase

MyoD:

Myoblast determination protein

NF-kB:

Nuclear factor activated B cells

NO:

Nitric oxide

nNOS:

Neuronal nitric oxide synthase

NOS:

Nitric oxide synthase

O2 − :

Superoxide

ONOO− :

Peroxynitrite

PL:

Plantaris muscle

ROS:

Radical oxygen species

SOL:

Soleus muscle

TOP2A:

Topoisomerase II

WG:

Medial gastrocnemius

References

  • Aldieri E, Bergandi L, Riganti C, Costamagna C, Bosia A, Ghigo D. Doxorubicin induces an increase of nitric oxide synthesis in rat cardiac cells that is inhibited by iron supplementation. Toxicol Appl Pharmacol. 2002;185:85–90.

    Article  CAS  PubMed  Google Scholar 

  • Antoun S, Baracos VE, Birdsell L, Escudier B, Sawyer MB. Low body mass index and sarcopenia associated with dose-limiting toxicity of sorafenib in patients with renal cell carcinoma. Ann Oncol. 2010;21:1594–8.

    Article  CAS  PubMed  Google Scholar 

  • Archer SL, Huang JM, Hampl V, Nelson DP, Shultz PJ, Weir EK. Nitric oxide and cGMP cause vasorelaxation by activation of a charybdotoxin-sensitive K channel by cGMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1994;91:7583–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Arola OJ, Saraste A, Pulkki K, Kallajoki M, Parvinen M, Voipio-Pulkki LM. Acute doxorubicin cardiotoxicity involves cardiomyocyte apoptosis. Cancer Res. 2000;60:1789–92.

    CAS  PubMed  Google Scholar 

  • Beckman JS, Beckman TW, Chen J, Marshall PA, Freeman BA. Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci U S A. 1990;87:1620–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Buck M, Chojkier M. Muscle wasting and dedifferentiation induced by oxidative stress in a murine model of cachexia is prevented by inhibitors of nitric oxide synthesis and antioxidants. EMBO J. 1996;15:1753–65.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Calabrese V, Mancuso C, Calvani M, Rizzarelli E, Butterfield DA, Stella AM. Nitric oxide in the central nervous system: neuroprotection versus neurotoxicity. Nat Rev Neurosci. 2007;8:766–75.

    Article  CAS  PubMed  Google Scholar 

  • Carter SK. Adriamycin-a review. J Natl Cancer Inst. 1975;55:1265–74.

    CAS  PubMed  Google Scholar 

  • Conklin KA. Chemotherapy-associated oxidative stress: impact on chemotherapeutic effectiveness. Integr Cancer Ther. 2004;3:294–300.

    Article  CAS  PubMed  Google Scholar 

  • De Palma C, Morisi F, Pambianco S, Assi E, Touvier T, Russo S, et al. Deficient nitric oxide signalling impairs skeletal muscle growth and performance: involvement of mitochondrial dysregulation. Skelet Muscle. 2014;4:22.

    Article  PubMed  PubMed Central  Google Scholar 

  • Delgado JM, Defeudis FV, Roth RH, Ryugo DK, Mitruka BM. Dialytrode for long term intracerebral perfusion in awake monkeys. Arch Int Pharmacodyn Ther. 1972;198:9–21.

    CAS  PubMed  Google Scholar 

  • Di Marco S, Mazroui R, Dallaire P, Chittur S, Tenenbaum SA, Radzioch D, et al. NF-kappa B-mediated MyoD decay during muscle wasting requires nitric oxide synthase mRNA stabilization, HuR protein, and nitric oxide release. Mol Cell Biol. 2005;25:6533–45.

    Article  PubMed  PubMed Central  Google Scholar 

  • Evig CB, Kelley EE, Weydert CJ, Chu Y, Buettner GR, Burns CP. Endogenous production and exogenous exposure to nitric oxide augment doxorubicin cytotoxicity for breast cancer cells but not cardiac myoblasts. Nitric Oxide. 2004;10:119–29.

    Article  CAS  PubMed  Google Scholar 

  • Fabris S, MacLean DA. Skeletal muscle an active compartment in the sequestering and metabolism of doxorubicin chemotherapy. PLoS One. 2015;10:e0139070.

    Article  PubMed  PubMed Central  Google Scholar 

  • Garner AP, Paine MJ, Rodriguez-Crespo I, Chinje EC, Ortiz de Montellano P, Stratford IJ, et al. Nitric oxide synthases catalyze the activation of redox cycling and bioreductive anticancer agents. Cancer Res. 1999;59:1929–34.

    CAS  PubMed  Google Scholar 

  • Gewirtz DA. A critical evaluation of the mechanisms of action proposed for the antitumor effects of the anthracycline antibiotics adriamycin and daunorubicin. Biochem Pharmacol. 1999;57:727–41.

    Article  CAS  PubMed  Google Scholar 

  • Gille L, Nohl H. Analyses of the molecular mechanism of adriamycin-induced cardiotoxicity. Free Radic Biol Med. 1997;23:775–82.

    Article  CAS  PubMed  Google Scholar 

  • Gilliam LA, St Clair DK. Chemotherapy-induced weakness and fatigue in skeletal muscle: the role of oxidative stress. Antioxid Redox Signal. 2011;15:2543–63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guerra J, De Jesus A, Santiago-Borrero P, Roman-Franco A, Rodriguez E, Crespo MJ. Plasma nitric oxide levels used as an indicator of doxorubicin-induced cardiotoxicity in rats. Hematol J. 2005;5:584–8.

    Article  CAS  PubMed  Google Scholar 

  • Hayward R, Hydock D, Gibson N, Greufe S, Bredahl E, Parry T. Tissue retention of doxorubicin and its effects on cardiac, smooth, and skeletal muscle function. J Physiol Biochem. 2013;69:177–87.

    Article  CAS  PubMed  Google Scholar 

  • Hellsten Y, MacLean D, Radegran G, Saltin B, Bangsbo J. Adenosine concentrations in the interstitium of resting and contracting human skeletal muscle. Circulation. 1998;98:6–8.

    Article  CAS  PubMed  Google Scholar 

  • Iqbal M, Dubey K, Anwer T, Ashish A, Pillai KK. Protective effects of telmisartan against acute doxorubicin-induced cardiotoxicity in rats. Pharmacol Rep. 2008;60:382–90.

    CAS  PubMed  Google Scholar 

  • Jackson MJ. Use of microdialysis to study interstitial nitric oxide and other reactive oxygen and nitrogen species in skeletal muscle. Methods Enzymol. 2005;396:514–25.

    Article  CAS  PubMed  Google Scholar 

  • Jensen RA, Acton EM, Peters JH. Doxorubicin cardiotoxicity in the rat: comparison of electrocardiogram, transmembrane potential, and structural effects. J Cardiovasc Pharmacol. 1984;6:186–200.

    Article  CAS  PubMed  Google Scholar 

  • Jung ID, Lee JS, Yun SY, Park CG, Han JW, Lee HW, et al. Doxorubicin inhibits the production of nitric oxide by colorectal cancer cells. Arch Pharm Res. 2002;25:691–6.

    Article  CAS  PubMed  Google Scholar 

  • Kalivendi SV, Kotamraju S, Zhao H, Joseph J, Kalyanaraman B. Doxorubicin-induced apoptosis is associated with increased transcription of endothelial nitric-oxide synthase. Effect of antiapoptotic antioxidants and calcium. J Biol Chem. 2001;276:47266–76.

    Article  CAS  PubMed  Google Scholar 

  • Kassner N, Huse K, Martin HJ, Godtel-Armbrust U, Metzger A, Meineke I, et al. Carbonyl reductase 1 is a predominant doxorubicin reductase in the human liver. Drug Metab Dispos. 2008;36:2113–20.

    Article  CAS  PubMed  Google Scholar 

  • Kotamraju S, Konorev EA, Joseph J, Kalyanaraman B. Doxorubicin-induced apoptosis in endothelial cells and cardiomyocytes is ameliorated by nitrone spin traps and ebselen. Role of reactive oxygen and nitrogen species. J Biol Chem. 2000;275:33585–92.

    Article  CAS  PubMed  Google Scholar 

  • Kremer LC, van Dalen EC, Offringa M, Ottenkamp J, Voute PA. Anthracycline-induced clinical heart failure in a cohort of 607 children: long-term follow-up study. J Clin Oncol. 2001;19:191–6.

    CAS  PubMed  Google Scholar 

  • Lim S, Shin JY, Jo A, Jyothi KR, Nguyen MN, Choi TG, et al. Carbonyl reductase 1 is an essential regulator of skeletal muscle differentiation and regeneration. Int J Biochem Cell Biol. 2013;45:1784–93.

    Article  CAS  PubMed  Google Scholar 

  • MacLean DA, Bangsbo J, Saltin B. Muscle interstitial glucose and lactate levels during dynamic exercise in humans determined by microdialysis. J Appl Physiol (1985). 1999;87:1483–90.

    CAS  Google Scholar 

  • MacLean DA, Vickery LM, Sinoway LI. Elevated interstitial adenosine concentrations do not activate the muscle reflex. Am J Physiol Heart Circ Physiol. 2001;280:H546–53.

    CAS  PubMed  Google Scholar 

  • Megeney LA, Kablar B, Garrett K, Anderson JE, Rudnicki MA. MyoD is required for myogenic stem cell function in adult skeletal muscle. Genes Dev. 1996;10:1173–83.

    Article  CAS  PubMed  Google Scholar 

  • Minotti G, Menna P, Salvatorelli E, Cairo G, Gianni L. Anthracyclines: molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity. Pharmacol Rev. 2004;56:185–229.

    Article  CAS  PubMed  Google Scholar 

  • Momparler RL, Karon M, Siegel SE, Avila F. Effect of adriamycin on DNA, RNA, and protein synthesis in cell-free systems and intact cells. Cancer Res. 1976;36:2891–5.

    CAS  PubMed  Google Scholar 

  • Nagi MN, Mansour MA. Protective effect of thymoquinone against doxorubicin-induced cardiotoxicity in rats: a possible mechanism of protection. Pharmacol Res. 2000;41:283–9.

    Article  CAS  PubMed  Google Scholar 

  • Octavia Y, Tocchetti CG, Gabrielson KL, Janssens S, Crijns HJ, Moens AL. Doxorubicin-induced cardiomyopathy: from molecular mechanisms to therapeutic strategies. J Mol Cell Cardiol. 2012;52:1213–25.

    Article  CAS  PubMed  Google Scholar 

  • Okutsu M, Call JA, Lira VA, Zhang M, Donet JA, French BA, et al. Extracellular superoxide dismutase ameliorates skeletal muscle abnormalities, cachexia, and exercise intolerance in mice with congestive heart failure. Circ Heart Fail. 2014;7:519–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Olson RD, Mushlin PS. Doxorubicin cardiotoxicity: analysis of prevailing hypotheses. FASEB J. 1990;4:3076–86.

    CAS  PubMed  Google Scholar 

  • Paulides M, Kremers A, Stohr W, Bielack S, Jurgens H, Treuner J, et al. Prospective longitudinal evaluation of doxorubicin-induced cardiomyopathy in sarcoma patients: a report of the late effects surveillance system (LESS). Pediatr Blood Cancer. 2006;46:489–95.

    Article  CAS  PubMed  Google Scholar 

  • Prado CM, Baracos VE, Mccargar LJ, Reiman T, Mourtzakis M, Tonkin K, et al. Sarcopenia as a determinant of chemotherapy toxicity and time to tumor progression in metastatic breast cancer patients receiving capecitabine treatment. Clin Cancer Res. 2009;15:2920–6.

    Article  CAS  PubMed  Google Scholar 

  • Ramamoorthy S, Donohue M, Buck M. Decreased Jun-D and myogenin expression in muscle wasting of human cachexia. Am J Physiol Endocrinol Metab. 2009;297:E392–401.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Riddell DR, Owen JS. Nitric oxide and platelet aggregation. Vitam Horm. 1999;57:25–48.

    Article  CAS  PubMed  Google Scholar 

  • Riede UN, Forstermann U, Drexler H. Inducible nitric oxide synthase in skeletal muscle of patients with chronic heart failure. J Am Coll Cardiol. 1998;32:964–9.

    Article  CAS  PubMed  Google Scholar 

  • Sawyer DB, Fukazawa R, Arstall MA, Kelly RA. Daunorubicin-induced apoptosis in rat cardiac myocytes is inhibited by dexrazoxane. Circ Res. 1999;84:257–65.

    Article  CAS  PubMed  Google Scholar 

  • Smuder AJ, Kavazis AN, Min K, Powers SK. Exercise protects against doxorubicin-induced markers of autophagy signaling in skeletal muscle. J Appl Physiol (1985). 2011;111:1190–8.

    Article  CAS  Google Scholar 

  • Stamler JS, Meissner G. Physiology of nitric oxide in skeletal muscle. Physiol Rev. 2001;81:209–37.

    CAS  PubMed  Google Scholar 

  • Teraoka K, Hirano M, Yamaguchi K, Yamashina A. Progressive cardiac dysfunction in adriamycin-induced cardiomyopathy rats. Eur J Heart Fail. 2000;2:373–8.

    Article  CAS  PubMed  Google Scholar 

  • van Norren K, van Helvoort A, Argiles JM, van Tuijl S, Arts K, Gorselink M, et al. Direct effects of doxorubicin on skeletal muscle contribute to fatigue. Br J Cancer. 2009;100:311–4.

    Article  PubMed  PubMed Central  Google Scholar 

  • Vasquez-Vivar J, Martasek P, Hogg N, Masters BS, Pritchard KA, Kalyanaraman B. Endothelial nitric oxide synthase-dependent superoxide generation from adriamycin. Biochemistry. 1997;36:11293–7.

    Article  CAS  PubMed  Google Scholar 

  • Wallimann T, Wyss M, Brdiczka D, Nicolay K, Eppenberger HM. Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands: the ‘phosphocreatine circuit’ for cellular energy homeostasis. Biochem J. 1992;281(Pt 1):21–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang S, Konorev EA, Kotamraju S, Joseph J, Kalivendi S, Kalyanaraman B. Doxorubicin induces apoptosis in normal and tumor cells via distinctly different mechanisms. intermediacy of H(2)O(2)- and p53-dependent pathways. J Biol Chem. 2004;279:25535–43.

    Article  CAS  PubMed  Google Scholar 

  • Xi L, Zhu SG, Das A, Chen Q, Durrant D, Hobbs DC, et al. Dietary inorganic nitrate alleviates doxorubicin cardiotoxicity: mechanisms and implications. Nitric Oxide. 2012;26:274–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yu AP, Pei XM, Sin TK, Yip SP, Yung BY, Chan LW, et al. Acylated and unacylated ghrelin inhibit doxorubicin-induced apoptosis in skeletal muscle. Acta Physiol (Oxf). 2014;211:201–13.

    Article  CAS  Google Scholar 

  • Zhang S, Liu X, Bawa-Khalfe T, Lu LS, Lyu YL, Liu LF, et al. Identification of the molecular basis of doxorubicin-induced cardiotoxicity. Nat Med. 2012;18:1639–42.

    Article  PubMed  Google Scholar 

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Acknowledgments

The authors wish to thank Ms. Pamela Chenard, Ms. Talia Ryan, and Ms. Josée Lalanne for their excellent technical assistance. This study was supported by the Natural Sciences and Engineering Research Council and the Ontario Institute for Cancer Research.

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Authors and Affiliations

  1. Biomolecular Sciences, Laurentian University, Sudbury, Ontario, Canada

    Sergio Fabris & David A. MacLean

  2. Division of Medical Sciences, Northern Ontario School of Medicine, Laurentian University Campus, 935 Ramsey Lake Road, Sudbury & Thunder Bay, Ontario, P3E 2C6, Canada

    David A. MacLean

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  1. Sergio Fabris
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Correspondence to Sergio Fabris.

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Fabris, S., MacLean, D.A. Doxorubicin chemotherapy affects intracellular and interstitial nitric oxide concentrations in skeletal muscle. Cell Biol Toxicol 32, 121–131 (2016). https://doi.org/10.1007/s10565-016-9325-1

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