Amino Acids

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Up-regulation of HIF-1α is associated with neuroprotective effects of agmatine against rotenone-induced toxicity in differentiated SH-SY5Y cells

  • Nadia Ferlazzo
  • Monica Currò
  • Maria Laura Giunta
  • Domenico Longo
  • Valentina Rizzo
  • Daniela Caccamo
  • Riccardo IentileEmail author
Original Article
Part of the following topical collections:
  1. Polyamines: Biochemical and Pathophysiological Properties


Agmatine, a metabolite generated by arginine decarboxylation, has been reported as neuromodulator and neuroactive substance. Several findings suggest that agmatine displays neuroprotective effects in several models of neurodegenerative disorders, such as Parkinson’s disease (PD). It has been hypothesized that biogenic amines may be involved in neuroprotection by scavenging oxygen radicals, thus preventing the generation of oxidative stress. Mitochondrial dysfunction, that leads to a reduction of oxygen consumption, followed by activation of prolyl hydroxylase and decrease of hypoxia-inducible factor 1 alpha (HIF-1α) levels, has been demonstrated to play a role in PD pathogenesis. Using rotenone-treated differentiated SH-SY5Y cells as the in vitro PD model, we here investigated the molecular mechanisms underlying agmatine neuroprotective effects. Our results showed that the preliminary addition of agmatine induces HIF-1α activation, and prevents the rotenone-induced production of free radical species, and the activation of apoptotic pathways by inhibiting mitochondrial membrane potential decrease and caspase 3 as well as cytochrome c increase. Notably, these effects are mediated by HIF-1α, as indicated by experiments using a HIF-1α inhibitor. The present findings suggest that the treatment with agmatine is able to counteract the neuronal cell injury evoked by mitochondrial toxins.


Agmatine Neuroprotective activity Hypoxia-inducible factor-1 Oxidative stress Apoptosis 



2′,7′-Dichlorofluorescein diacetate


All-trans retinoic acid


Eagle’s minimum essential medium


Fetal bovine serum


Ham’s F-12 nutrient mixture


Hypoxia-inducible factor-1


3-(4,5-Methylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide


Parkinson’s disease


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

All authors listed have contributed to the conception, design, gathering, analysis or interpretation of data and have contributed to the writing and intellectual content of the article. All authors gave informed consent to the submission of this manuscript.


  1. Arndt MA, Battaglia V, Parisi E, Lortie MJ, Isome M, Baskerville C, Pizzo DP, Ientile R, Colombatto S, Toninello A, Satriano J (2009) The arginine metabolite agmatine protects mitochondrial function and confers resistance to cellular apoptosis. Am J Physiol Cell Physiol 296:C1411–C1419. CrossRefGoogle Scholar
  2. Bader HL, Hsu T (2012) Systemic VHL gene functions and the VHL disease. FEBS Lett 586:1562–1569. CrossRefGoogle Scholar
  3. Battaglia V, Rossi CA, Colombatto S, Grillo MA, Toninello A (2007) Different behavior of agmatine in liver mitochondria: inducer of oxidative stress or scavenger of reactive oxygen species? Biochim Biophys Acta 1768:1147–1153. CrossRefGoogle Scholar
  4. Battaglia V, Grancara S, Satriano J, Saccoccio S, Agostinelli E, Toninello A (2010a) Agmatine prevents the Ca(2+)-dependent induction of permeability transition in rat brain mitochondria. Amino Acids 38:431–437. CrossRefGoogle Scholar
  5. Battaglia V, Grancara S, Mancon M, Cravanzola C, Colombatto S, Grillo MA, Tempera G, Agostinelli E, Toninello A (2010b) Agmatine transport in brain mitochondria: a different mechanism from that in liver mitochondria. Amino Acids 38(2):423–430. CrossRefGoogle Scholar
  6. Caccamo D, Campisi A, Currò M, Bramanti V, Tringali M, Li Volti G, Vanella A, Ientile R (2005) Antioxidant treatment inhibited glutamate-evoked NF-kappaB activation in primary astroglial cell cultures. Neurotoxicology 26:915–921. CrossRefGoogle Scholar
  7. Condello S, Currò M, Ferlazzo N, Caccamo D, Satriano J, Ientile R (2011) Agmatine effects on mitochondrial membrane potential and NF-κB activation protect against rotenone-induced cell damage in human neuronal-like SH-SY5Y cells. J Neurochem 116:67–75. CrossRefGoogle Scholar
  8. Correia SC, Moreira PI (2010) Hypoxia-inducible factor 1: a new hope to counteract neurodegeneration? J Neurochem 112:1–12. CrossRefGoogle Scholar
  9. Ferlazzo N, Condello S, Currò M, Parisi G, Ientile R, Caccamo D (2008) NF-kappaB activation is associated with homocysteine-induced injury in Neuro2a cells. BMC Neurosci 9:62. CrossRefGoogle Scholar
  10. Galanis A, Pappa A, Giannakakis A, Lanitis E, Dangaj D, Sandaltzopoulos R (2008) Reactive oxygen species and HIF-1 signalling in cancer. Cancer Lett 266:12–20. CrossRefGoogle Scholar
  11. Gardini G, Cabella C, Cravanzola C, Vargiu C, Belliardo S, Testore G, Solinas SP, Toninello A, Grillo MA, Colombatto S (2001) Agmatine induces apoptosis in rat hepatocyte cultures. J Hepatol 35:482–489CrossRefGoogle Scholar
  12. Gilad GM, Gilad VH (2000) Accelerated functional recovery and neuroprotection by agmatine after spinal cord ischemia in rats. Neurosci Lett 296:97–100CrossRefGoogle Scholar
  13. Gorbatyuk OS, Milner TA, Wang G, Regunathan S, Reis DJ (2001) Localization of agmatine in vasopressin and oxytocin neurons of the rat hypothalamic paraventricular and supraoptic nuclei. Exp Neurol 171:235–245. CrossRefGoogle Scholar
  14. Guo S, Miyake M, Liu KJ, Shi H (2009) Specific inhibition of hypoxia inducible factor 1 exaggerates cell injury induced by in vitro ischemia through deteriorating cellular redox environment. J Neurochem 108:1309–1321. CrossRefGoogle Scholar
  15. Halaris A, Plietz J (2007) Agmatine: metabolic pathway and spectrum of activity in brain. CNS Drugs 21:885–900CrossRefGoogle Scholar
  16. Hong S, Kim CY, Lee JE, Seong GJ (2009) Agmatine protects cultured retinal ganglion cells from tumor necrosis factor-alpha-induced apoptosis. Life Sci 84:28–32. CrossRefGoogle Scholar
  17. Horyn O, Luhovyy B, Lazarow A, Daikhin Y, Nissim I, Yudkoff M, Nissim I (2005) Biosynthesis of agmatine in isolated mitochondria and perfused rat liver: studies with 15N-labelled arginine. Biochem J 388:419–425. CrossRefGoogle Scholar
  18. Hutt DM, Roth DM, Vignaud H, Cullin C, Bouchecareilh M (2014) The histone deacetylase inhibitor, vorinostat, represses hypoxia inducible factor 1 alpha expression through translational inhibition. PLoS One 9:e106224. CrossRefGoogle Scholar
  19. Iizuka Y, Hong S, Kim CY, Yang WI, Lee JE, Seong GJ (2010) Protective mechanism of agmatine pretreatment on RGC-5 cells injured by oxidative stress. Braz J Med Biol Res 43(4):356–358. CrossRefGoogle Scholar
  20. Ke Q, Costa M (2006) Hypoxia-inducible factor-1 (HIF-1). Mol Pharmacol 70:1469–1480. CrossRefGoogle Scholar
  21. Kim JH, Yenari MA, Giffard RG, Cho SW, Park KA, Lee JE (2004) Agmatine reduces infarct area in a mouse model of transient focal cerebral ischemia and protects cultured neurons from ischemia-like injury. Exp Neurol 189:122–130. CrossRefGoogle Scholar
  22. Langston JW, Ballard P, Tetrud JW, Irwin I (1983) Chronic Parkinsonism in humans due to a product of meperidine-analog synthesis. Science 219:979–980CrossRefGoogle Scholar
  23. Lortie MJ, Novotny WF, Peterson OW, Vallon V, Malvey K, Mendonca M, Satriano J, Insel P, Thomson SC, Blantz RC (1996) Agmatine, a bioactive metabolite of arginine. Production, degradation, and functional effects in the kidney of the rat. J Clin Investig 97:13–420. CrossRefGoogle Scholar
  24. Nissim I, Horyn O, Daikhin Y, Nissim I, Lazarow A, Yudkoff M (2002) Regulation of urea synthesis by agmatine in the perfused liver: studies with 15N. Am J Physiol Endocrinol Metab 283:E1123–E1134. CrossRefGoogle Scholar
  25. Permenter MG, Dennis WE, Sutto TE, Jackson DA, Lewis JA, Stallings JD (2013) Exposure to cobalt causes transcriptomic and proteomic changes in two rat liver derived cell lines. PLoS One 8:e83751. CrossRefGoogle Scholar
  26. Regunathan S, Reis DJ (2000) Characterization of arginine decarboxylase in rat brain and liver: distinction from ornithine decarboxylase. J Neurochem 74:2201–2208CrossRefGoogle Scholar
  27. Roberts JC, Grocholski BM, Kitto KF, Fairbanks CA (2005) Pharmacodynamic and pharmacokinetic studies of agmatine after spinal administration in the mouse. J Pharmacol Exp Ther 314:1226–1233. CrossRefGoogle Scholar
  28. Salminen A, Kaarniranta K, Kauppinen A (2016) AMPK and HIF signaling pathways regulate both longevity and cancer growth: the good news and the bad news about survival mechanisms. Biogerontology 17:655–680. CrossRefGoogle Scholar
  29. Sharp FR, Bernaudin M (2004) HIF1 and oxygen sensing in the brain. Nat Rev Neurosci 5:437–448. CrossRefGoogle Scholar
  30. Siddiq A, Ayoub IA, Chavez JC, Aminova L, Shah S, LaManna JC, Patton SM, Connor JR, Cherny RA, Volitakis I, Bush AI, Langsetmo I, Seeley T, Gunzler V, Ratan RR (2005) Hypoxia-inducible factor prolyl 4-hydroxylase inhibition. A target for neuroprotection in the central nervous system. J Biol Chem 280:41732–41743. CrossRefGoogle Scholar
  31. Sluch VM, Davis CH, Ranganathan V, Kerr JM, Krick K, Martin R, Berlinicke CA, Marsh-Armstrong N, Diamond JS, Mao HQ, Zack DJ (2015) Differentiation of human ESCs to retinal ganglion cells using a CRISPR engineered reporter cell line. Sci Rep 5:16595. CrossRefGoogle Scholar
  32. Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M (1997) Alpha-synuclein in Lewy bodies. Nature 388:839–840. CrossRefGoogle Scholar
  33. Sugiura T, Tsutsui H, Takaoka M, Kobuchi S, Hayashi K, Fujii T, Matsumura Y (2008) Protective effect of agmatine on ischemia/reperfusion-induced renal injury in rats. J Cardiovasc Pharmacol 51:223–230. CrossRefGoogle Scholar
  34. Wang GL, Jiang BH, Rue EA, Semenza GL (1995) Hypoxia-inducible factor 1 is a basic-helix–loop–helix-PAS heterodimer regulated by cellular O2 tension. Proc Natl Acad Sci USA 92:5510–5514CrossRefGoogle Scholar
  35. Wang WP, Iyo AH, Miguel-Hidalgo J, Regunathan S, Zhu MY (2006) Agmatine protects against cell damage induced by NMDA and glutamate in cultured hippocampal neurons. Brain Res 1084:210–216. CrossRefGoogle Scholar
  36. Winklhofer KF, Haass C (2010) Mitochondrial dysfunction in Parkinson’s disease. Biochim Biophys Acta 1802:29–44. CrossRefGoogle Scholar
  37. Wu G, Bazer FW, Davis TA, Kim SW, Li P, Marc Rhoads J, Carey Satterfield M, Smith SB, Spencer TE, Yin Y (2009) Arginine metabolism and nutrition in growth, health and disease. Amino Acids 37:153–168. CrossRefGoogle Scholar
  38. Wu Y, Li X, Xie W, Jankovic J, Le W, Pan T (2010) Neuroprotection of deferoxamine on rotenone-induced injury via accumulation of HIF-1 alpha and induction of autophagy in SH-SY5Y cells. Neurochem Int 57:198–205. CrossRefGoogle Scholar
  39. Xin XY, Pan J, Wang XQ, Ma JF, Ding JQ, Yang GY, Chen SD (2011) 2-methoxyestradiol attenuates autophagy activation after global ischemia. Can J Neurol Sci J Can Sci Neurol 38:631–638CrossRefGoogle Scholar
  40. Yang XC, Reis DJ (1999) Agmatine selectively blocks the N-methyl-d-aspartate subclass of glutamate receptor channels in rat hippocampal neurons. J Pharmacol Exp Ther 288:544–549Google Scholar
  41. Youdim MBH, Kupershmidt L, Amit T, Weinreb O (2014) Promises of novel multi-target neuroprotective and neurorestorative drugs for Parkinson’s disease. Parkinsonism Relat Disord 20:S132–S136. CrossRefGoogle Scholar
  42. Zhu MY, Piletz JE, Halaris A, Regunathan S (2003) Effect of agmatine against cell death induced by NMDA and glutamate in neurons and PC12 cells. Cell Mol Neurobiol 23:865–872CrossRefGoogle Scholar
  43. Zhu MY, Wang WP, Cai ZW, Regunathan S, Ordway G (2008) Exogenous agmatine has neuroprotective effects against restraint-induced structural changes in the rat brain. Eur J Neurosci 27:1320–1332. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Nadia Ferlazzo
    • 1
  • Monica Currò
    • 1
  • Maria Laura Giunta
    • 1
  • Domenico Longo
    • 1
  • Valentina Rizzo
    • 1
  • Daniela Caccamo
    • 1
  • Riccardo Ientile
    • 1
    Email author
  1. 1.Department of Biomedical and Dental Sciences and Morphofunctional ImagingUniversity of Messina, Policlinico G. MartinoMessinaItaly

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