Skip to main content
SpringerLink
Log in

Protective Effect of PACAP Against Doxorubicin-Induced Cell Death in Cardiomyocyte Culture

  • Published:
Journal of Molecular Neuroscience Aims and scope Submit manuscript

Abstract

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a widely distributed endogenous neuropeptide, also occurring in the cardiovascular system. Among others, PACAP has been suggested as a cardioprotective factor. It has been shown that PACAP inhibits cardiac fibrosis and protects cardiomyocytes against oxidative stress and in vitro ischemia/reperfusion. The aim of the present study was to investigate whether PACAP is protective in doxorubicin-induced cell death of cardiomyocytes. Cardiomyocytes were exposed to 1 µM doxorubicin for 24 h, which resulted in a marked reduction of cell viability and a parallel increase of apoptotic cells assessed by MTT test and annexin V/propidium iodide flow cytometry assay. Co-incubation with 20 nM PACAP increased cell viability and reduced the percentage of apoptotic cells. Furthermore, doxorubicin increased the activation of caspase-3 and decreased the phosphorylation of Bad, while simultaneous PACAP treatment reduced the caspase-3 activation and increased the level of phospho-Bad. In summary, our present results demonstrate that PACAP effectively protects cardiomyocytes against doxorubicin-induced apoptotic cell death.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  • Baron A, Monnier D, Roatti A, Baertschi AJ (2001) Pituitary adenylate cyclase activating polypeptide activates K(ATP) current in rat atrial myocytes. Am J Physiol Heart Circ Physiol 280:H1058–H1065

    CAS  PubMed  Google Scholar 

  • Berdichevski A, Meiry G, Milman F et al (2010) TVP1022 protects neonatal rat ventricular myocytes against doxorubicin-induced functional derangements. J Pharmacol Exp Ther 332:413–420

    Article  CAS  PubMed  Google Scholar 

  • Braas KM, Rossignol TM, Girard BM, May V, Parsons RL (2004) Pituitary adenylate cyclase activating polypeptide (PACAP) decreases neuronal somatostatin immunoreactivity in cultured guinea-pig parasympathetic cardiac ganglia. Neuroscience 126:335–346

    Article  CAS  PubMed  Google Scholar 

  • Bruch L, Bychkov R, Kastner A et al (1997) Pituitary adenylate cyclase activating peptides relax human coronary arteries by activating K(ATP) and K(Ca) channels in smooth muscle. J Vasc Res 34:11–18

    CAS  PubMed  Google Scholar 

  • Chang Y, Lawson LJ, Hancock JC, Hoover DB (2005) Pituitary adenylate cyclase activating polypeptide: localization and differential influence on isolated hearts from rats and gunea pigs. Regul Pept 129:139–146

    Article  CAS  PubMed  Google Scholar 

  • Chen QM, Tu VC (2002) Apoptosis and heart failure: mechanisms and therapeutic implications. Am J Cardiovasc Drugs 2:43–57

    Article  CAS  PubMed  Google Scholar 

  • Dalsgaard T, Hannibal J, Fahrenkrug J, Larsen CR, Ottesen B (2003) VIP and PACAP display different vasodilatory effects in rabbit coronary and cerebral arteries. Regul Pept 110:179–188

    Article  CAS  PubMed  Google Scholar 

  • Dejda A, Jolivel V, Bourgault S et al (2008) Inhibitory effect of PACAP on caspase activity in neuronal apoptosis: a better understanding towards therapeutic applications in neurodegenerative diseases. J Mol Neurosci 36:26–37

    Article  CAS  PubMed  Google Scholar 

  • Delgado M, Ganea D (2000) VIP and PACAP inhibit activation induced apoptosis in T lymphocytes. Ann N Y Acad Sci 921:55–67

    Article  CAS  PubMed  Google Scholar 

  • Dvorakova MC (2005) Cardioprotective role of the VIP signaling system. Drugs News Perspect 18:387–391

    Article  CAS  Google Scholar 

  • Falluel-Morel A, Aubert N, Vaudry D et al (2008) Interactions of PACAP and ceramides in the control of granule cell apoptosis during cerebellar development. J Mol Neurosci 36:8–15

    Article  CAS  PubMed  Google Scholar 

  • Gasz B, Racz B, Roth E et al (2006a) PACAP inhibits oxidative stress-induced activation of MAP kinase dependent apoptotic pathway in cultured cardiomyocytes. Ann N Y Acad Sci 1070:293–297

    Article  CAS  PubMed  Google Scholar 

  • Gasz B, Racz B, Roth E et al (2006b) Pituitary adenylate cyclase activating polypeptide protects cardiomyocytes against oxidative stress-induced apoptosis. Peptides 27:87–94

    Article  CAS  PubMed  Google Scholar 

  • Gilleron M, Marechal X, Montaigne D, Franczak J, Neviere R, Lancel S (2009) NADPH oxidases participate to doxorubicin-induced cardiac myocyte apoptosis. Biochem Biophys Res Commun 388:727–731

    Article  CAS  PubMed  Google Scholar 

  • Gutierrez-Canas I, Rodriguez-Henche N, Bolanos O, Carmena MJ, Prieto JC, Juarranz MG (2003) VIP and PACAP are autocrine factors that protect the androgen-independent prostate cancer cell line PC-3 from apoptosis induced by serum withdrawal. Br J Pharmacol 139:1050–1058

    Article  CAS  PubMed  Google Scholar 

  • Han P, Lucero MT (2005) Pituitary adenylate cyclase activating polypeptide reduces A-type K+ currents and caspase activity in cultured adult mouse olfactory neurons. Neuroscience 134:745–756

    Article  CAS  PubMed  Google Scholar 

  • Han X, Ren D, Fan P, Shen T, Lou H (2008) Protective effects of naringenin-7-O-glucoside on doxorubicin-induced apoptosis in H9C2 cells. Eur J Pharmacol 581:47–53

    Article  CAS  PubMed  Google Scholar 

  • Hirose M, Chiba S (2003) Cellular mechanism of pituitary adenylate cyclase activating polypeptide-induced atrial tachyarrhythmia in canince isolated arterially perfused right atria. Clin Exp Pharmacol Physiol 30:937–942

    Article  CAS  PubMed  Google Scholar 

  • Jantas D, Lason W (2009) Protective effect of memantine against doxorubicin toxicity in primary neuronal cell cultures: influence a developmental stage. Neurotox Res 15:24–37

    Article  CAS  PubMed  Google Scholar 

  • Konorev EA, Vanamala S, Kalyanaraman B (2008) Differences in doxorubicin-induced apoptotic signaling in adult and immature cardiomyocytes. Free Radic Biol Med 45:1723–1728

    Article  CAS  PubMed  Google Scholar 

  • Lee J, Park HJ, Choi HS et al (1999) Gonadotropin stimulation of pituitary adenylate cyclase activating polypeptide (PACAP) messenger ribonucleic acid in the rat ovary and the role of PACAP as a follicle survival factor. Endocrinology 140:818–826

    Article  CAS  PubMed  Google Scholar 

  • Lenti L, Domoki F, Kis D et al (2007) Pituitary adenylate cyclase activating polypeptide induces pial arteriolar vasodilation through cyclooxigenase-dependent and independent mechanisms in newborn pigs. Brain Res 1165:18–88

    Article  Google Scholar 

  • Lenti L, Zimmermann A, Kis D et al (2009) PACAP and VIP differentially preserve neurovascular reactivity after global cerebral ischemia in newborn pigs. Brain Res 1283:50–57

    Article  CAS  PubMed  Google Scholar 

  • Li M, David C, Kikuta T, Somogyvari-Vigh A, Arimura A (2005) Signaling cascades involved in neuroprotection by subpicomolar pituitary adenylate cyclase activating polypeptide 38. J Mol Neurosci 27:91–105

    Article  PubMed  Google Scholar 

  • Liu DM, Cuevas J, Adams DJ (2000) VIP and PACAP potentiation of nicotinic Ach-evoked currents in rat parasympathetic neurons is mediated by G-protein activation. Eur J NeuroSci 12:2243–2251

    Article  CAS  PubMed  Google Scholar 

  • Luodonpa M, Vuolteenaho O, Eskelinen S, Ruskoaho H (2001) Effects of adrenomedullin on hypertrophic responses induced by angiotensin II, endothelin-1 and phenylephrine. Peptides 22:1859–1866

    Article  Google Scholar 

  • Mori H, Nakamachi T, Ohtaki H et al (2009) Endogenous PACAP as a cardioprotectant in Doxorubicin-induced cardiomyopathy model in rats. Abstract NoP15, Presented at Phylogenetic Aspects of Neuropeptides/from Invertebrates to Humans, Satellite Symposium of the 9th International Symposium on VIP, PACAP and Related Peptides, Yakushima, Japan, 2009

  • Oka H, Jin L, Kulig E, Scheithauer BW, Lloyd RV (1999) Pituitary adenylate cyclase activating polypeptide inhibits transforming growth factor-beta 1-induced apoptosis in a human pituitary adenoma cell line. Am J Pathol 155:1893–1900

    CAS  PubMed  Google Scholar 

  • Otto C, Hein L, Brede M et al (2004) Pulmonary hypertension and right heart failure in pituitary adenylate cyclase activating polypeptide type I receptor-deficient mice. Circulation 110:3245–3251

    Article  CAS  PubMed  Google Scholar 

  • Pirger Z, Nemeth J, Hiripi L et al (2008) PACAP has antiapoptotic effect in the salivary gland of an invertebrate species, Helix pomatia. J Mol Neurosci 36:105–114

    Article  CAS  PubMed  Google Scholar 

  • Pugh PC, Margiotta JF (2006) PACAP support of neuronal survival requires MAPK- and activity-generated signals. Mol Cell Neurosci 31:586–595

    Article  CAS  PubMed  Google Scholar 

  • Racz B, Gasz B, Borsiczky B et al (2007a) Protective effects of pituitary adenylate cyclase activating polypeptide in endothelial cells against oxidative stress-induced apoptosis. Gen Comp Endocrinol 153:115–123

    Article  CAS  PubMed  Google Scholar 

  • Racz B, Gallyas F Jr, Kiss P et al (2007b) Effects of pituitary adenylate cyclase activating polypeptide (PACAP) on the PKA-Bad-14-3-3 signaling pathway in glutamate-induced retinal injury in neonatal rats. Neurotox Res 12:95–104

    Article  CAS  PubMed  Google Scholar 

  • Racz B, Gasz B, Gallyas F Jr et al (2008) PKA-Bad-14-3-3 and Akt-Bad-14-3-3 signaling pathways are involved in the protective effects of PACAP against ischemia/reperfusion-induced cardiomyocyte apoptosis. Regul Pept 145:105–115

    Article  CAS  PubMed  Google Scholar 

  • Racz B, Horvath G, Reglodi D et al (2010) PACAP ameliorates oxidative stress in the chicken inner ear: an in vitro study. Regul Pept 160:91–98

    Article  CAS  PubMed  Google Scholar 

  • Ross-Ascuitto NT, Ascuitto RJ, Ramage D, Kydon DW, Coy DH, Kadowitz PJ (1993) Pituitary adenylate cyclase activating polypeptide: a neuropeptide with potent inotropic and coronary vasodilatory effects in neonatal pig heart. Pediatr Res 34:323–328

    Article  CAS  PubMed  Google Scholar 

  • Roth E, Weber G, Kiss P et al (2009) Effects of PACAP and preconditioning against ischemia/reperfusion-induced cardiomyocyte apoptosis in vitro. Ann N Y Acad Sci 1163:512–516

    Article  CAS  PubMed  Google Scholar 

  • Sanchez A, Rao HV, Grammas P (2009) PACAP38 protects rat cortical neurons against neurotoxicity evoked by sodium nitroprusside and thrombin. Regul Pept 152:33–40

    Article  CAS  PubMed  Google Scholar 

  • Sano H, Miyata A, Horio T, Nishikimi T, Matsuo H, Kangawa K (2002) The effect of pituitary adenylate cyclase activating polypeptide on cultured rat cardiocytes as a cardioprotective factor. Regul Pept 109:107–113

    Article  CAS  PubMed  Google Scholar 

  • Schoenfeld LK, Souder JA, Hardwick JC (2000) Pituitary adenylate cyclase activating polypeptide innervation of the mudpuppy cardiac ganglion. Brain Res 882:180–190

    Article  CAS  PubMed  Google Scholar 

  • Somogyvari-Vigh A, Reglodi D (2004) Pituitary adenylate cyclase activating polypeptide: a potential neuroprotective peptide-review. Curr Pharm Des 10:2861–2889

    Article  CAS  PubMed  Google Scholar 

  • Suzuki Y, Kasai K, Takekoshi K et al (1993) Effects of pituitary adenylate cyclase activating polypeptide (PACAP) on the cardiovascular system. Regul Pept 47:213–220

    Article  CAS  PubMed  Google Scholar 

  • Tokola H, Salo K, Vuolteenaho O, Ruskoaho H (1994) Basal and acidic fibroblast growth factor-induced atrial natriuretic peptide gene expression and secretion is inhibited by staurosporine. Eur J Pharmacol 267:195–206

    Article  CAS  PubMed  Google Scholar 

  • Tompkins JD, Parsons RL (2008) Identification of intracellular signaling cascades mediating the PACAP-induced increase in guinea pig cardiac neuron excitability. J Mol Neurosci 36:292–298

    Article  CAS  PubMed  Google Scholar 

  • Vaudry D, Gonzalez BJ, Basille M et al (2000) The neuroprotective effect of pituitary adenylate cyclase activating polypeptide on cerebellar granule cells is mediated through inhibition of the CED3-related cysteine protease caspase-3/CPP32. Proc Natl Acad Sci USA 97:13390–13395

    Article  CAS  PubMed  Google Scholar 

  • Vaudry D, Falluel-Morel A, Bourgault A et al (2009) Pituitary adenylate cyclase activating polypeptide and its receptors: 20 years after the discovery. Pharmacol Rev 61:283–357

    Article  PubMed  Google Scholar 

  • Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C (1995) A novel assay for apoptosis. Flow cytometric detection of phospatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. J Immunol Methods 184:39–51

    Article  CAS  PubMed  Google Scholar 

  • Wang G, Qi C, Fan GH, Zhou HY, Chen SD (2005) PACAP protects neuronal differentiated PC12 cells against the neurotoxicity induced by a mitochondrial complex I inhibitor, rotenone. FEBS Lett 579:4005–4011

    Article  CAS  PubMed  Google Scholar 

  • Weiss RB (1992) The anthracyclines: will we ever find a better doxorubicin? Semin Oncol 19:670–686

    CAS  PubMed  Google Scholar 

  • Zaccone G, Mauceri A, Maisano M, Giannetto A, Parrino V, Fasulo S (2009) Distribution and neurotransmitter localization in the heart of the ray-finned fish, bichir (Polypterus bichir bichir Geoffroy St. Hilare, 1802). Acta Histochem 111:93–103

    Article  CAS  PubMed  Google Scholar 

  • Zhang Y, Liu LP, Liang ZL, Li XL, Jin YZ, Cui X (2008) cAMP produced by pituitary adenylate cyclase activating polypeptide 27 inhibits atrial natriuretic peptide secretion in rabbit beating atria. Clin Exp Pharmacol Physiol 35:1233–1237

    Article  CAS  PubMed  Google Scholar 

  • Zhang SH, Wang WQ, Wang JL (2009) Protective effects of tetrahydroxystilbene glucoside on cardiotoxicity induced by doxorubicin in vitro and in vivo. Acta Pharmacologica Sinica 30:1479–1487

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the Hungarian Science Research Fund OTKA K72592, K78434, CNK 78480, ETT278-04/2009, Bolyai Scholarship.

Author information

Authors and Affiliations

  1. Department of Biochemistry and Medical Chemistry, University of Pecs, Szigeti u 12, 7624, Pecs, Hungary

    Boglarka Racz & Andras Szigeti

  2. Hungary Department of Anatomy, University of Pecs, Pecs, Hungary

    Dora Reglodi & Gabriella Horvath

  3. Department of Surgical Research and Techniques, University of Pecs, Pecs, Hungary

    Borbala Balatonyi, Erzsebet Roth & Gyorgy Weber

  4. Department of Cardiac Surgery, Zala County Hospital, Zalaegerszeg, Hungary

    Nasri Alotti & Balazs Gasz

  5. Department of Medical Chemistry, University of Szeged, Szeged, Hungary

    Gabor Toth

Authors
  1. Boglarka Racz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dora Reglodi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gabriella Horvath
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Andras Szigeti
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Borbala Balatonyi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Erzsebet Roth
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Gyorgy Weber
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Nasri Alotti
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Gabor Toth
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Balazs Gasz
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Boglarka Racz.

Additional information

Boglarka Racz and Dora Reglodi made equal contribution to the present work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Racz, B., Reglodi, D., Horvath, G. et al. Protective Effect of PACAP Against Doxorubicin-Induced Cell Death in Cardiomyocyte Culture. J Mol Neurosci 42, 419–427 (2010). https://doi.org/10.1007/s12031-010-9349-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12031-010-9349-6

Keywords

Search

Navigation