Advertisement

Molecular and Cellular Biochemistry

, Volume 409, Issue 1–2, pp 199–211 | Cite as

Neuroprotective efficacy of naringin on 3-nitropropionic acid-induced mitochondrial dysfunction through the modulation of Nrf2 signaling pathway in PC12 cells

  • Gopinath Kulasekaran
  • Sudhandiran Ganapasam
Article

Abstract

Oxidative stress and mitochondrial dysfunction are implicated in neuronal apoptosis associated with Huntington’s disease. Naringin is the flavanone present in grapefruit and related citrus species possess diverse pharmacological and therapeutic properties including antioxidant, anti-apoptotic, and neuroprotective properties. The aim of this study was to investigate the protective effect of naringin on 3-nitropropionic acid (3-NP)-induced neurotoxicity in pheochromocytoma cells (PC12) cells and to explore its mechanism of action. Naringin protects PC12 cells from 3-NP neurotoxicity, as evaluated the by cell viability assays. The lactate dehydrogenase release was decreased upon naringin treatment in 3-NP-induced PC12 cells. Naringin treatment enhances the antioxidant defense by increasing the activities of enzymatic antioxidants and the level of reduced glutathione. The increase in levels of reactive oxygen species and lipid peroxidation induced by 3-NP were significantly decreased by naringin. PC12 cells induced with 3-NP showed decrease in the mitochondrial membrane potential and mitochondrial respiratory complex enzymes, succinate dehydrogenase and cytochrome c oxidase activities, and it was significantly altered to near normal upon naringin treatment. Naringin reduced the 3-NP-induced apoptosis through the modulation in expressions of B-cell lymphoma 2 and Bcl-2-associated X protein. Further, naringin enhances the nuclear translocation of Nrf2 and induces the NAD(P)H:quinone oxidoreductase-1 and Heme oxygenase-1 expressions through the phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathway. Taken together, the above findings suggest that naringin augments cellular antioxidant defense capacity and reduces the 3-NP-induced neurotoxicity in PC12 cells through the PI-3K/Akt-dependent Nrf2 activation in PC12 cells.

Keywords

Naringin 3-Nitropropionic acid Oxidative stress Mitochondrial dysfunction Apoptosis Nrf2 PI-3K/Akt 

Notes

Acknowledgments

We thank Lady Tata Memorial Trust, Mumbai, India and Council of Scientific and Industrial Research, New Delhi, India for financial assistance awarded to KG.

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.

References

  1. 1.
    Huang LS, Sun G, Cobessi D, Wang AC, Shen JT, Tung EY, Anderson VE, Berry EA (2006) 3-Nitropropionic acid is a suicide inhibitor of mitochondrial respiration that, upon oxidation by complex II, forms a covalent adduct with a catalytic base arginine in the active site of the enzyme. J Biol Chem 281:5965–5972PubMedCentralCrossRefPubMedGoogle Scholar
  2. 2.
    Shivasharan BD, Nagakannan P, Thippeswamy BS, Veerapur VP, Bansal P, Unnikrishnan MK (2013) Protective effect of Calendula officinalis Linn. flowers against 3-nitropropionic acid induced experimental Huntington’s disease in rats. Drug Chem Toxicol 36:466–473CrossRefPubMedGoogle Scholar
  3. 3.
    Hannan AJ (2005) Novel therapeutic targets for Huntington’s disease. Expert Opin Ther Targets 9:639–650CrossRefPubMedGoogle Scholar
  4. 4.
    Tsang TM, Haselden JN, Holmes E (2009) Metabonomic characterization of the 3-nitropropionic acid rat model of Huntington’s disease. Neurochem Res 34:1261–1271CrossRefPubMedGoogle Scholar
  5. 5.
    Tasset I, Pontes AJ, Hinojosa AJ, de la Torre R, Túnez I (2011) Olive oil reduces oxidative damage in a 3-nitropropionic acid-induced Huntington’s disease-like rat model. Nutr Neurosci 14:106–111CrossRefPubMedGoogle Scholar
  6. 6.
    Federico A, Cardaioli E, Da Pozzo P, Formichi P, Gallus GN, Radi E (2012) Mitochondria, oxidative stress and neurodegeneration. J Neurol Sci 322:254–262CrossRefPubMedGoogle Scholar
  7. 7.
    Damiano M, Diguet E, Malgorn C, D’Aurelio M, Galvan L, Petit F, Benhaim L, Guillermier M, Houitte D, Dufour N, Hantraye P, Canals JM, Alberch J, Delzescaux T, Déglon N, Beal MF, Brouillet E (2013) A role of mitochondrial complex II defects in genetic models of Huntington’s disease expressing N-terminal fragments of mutant huntingtin. Hum Mol Genet 22:3869–3882PubMedCentralCrossRefPubMedGoogle Scholar
  8. 8.
    Lin MT, Beal MF (2006) Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. Nature 443:787–795CrossRefPubMedGoogle Scholar
  9. 9.
    Browne SE, Ferrante RJ, Beal MF (1999) Oxidative stress in Huntington’s disease. Brain Pathol 9:147–163CrossRefPubMedGoogle Scholar
  10. 10.
    Quintanilla RA, Johnson GV (2009) Role of mitochondrial dysfunction in the pathogenesis of Huntington’s disease. Brain Res Bull 80:242–247PubMedCentralCrossRefPubMedGoogle Scholar
  11. 11.
    Hariharan A, Shetty S, Shirole T, Jagtap AG (2014) Potential of protease inhibitor in 3-nitropropionic acid induced Huntington’s disease like symptoms: mitochondrial dysfunction and neurodegeneration. Neurotoxicology 45:139–148CrossRefPubMedGoogle Scholar
  12. 12.
    Baird L, Dinkova-Kostova AT (2011) The cytoprotective role of the Keap1–Nrf2 pathway. Arch Toxicol 85:241–272CrossRefPubMedGoogle Scholar
  13. 13.
    de Vries HE, Witte M, Hondius D, Rozemuller AJ, Drukarch B, Hoozemans J, van Horssen J (2008) Nrf2-induced antioxidant protection: a promising target to counteract ROS-mediated damage in neurodegenerative disease? Free Radic Biol Med 45:1375–1383CrossRefPubMedGoogle Scholar
  14. 14.
    Kumar H, Koppula S, Kim IS, More SV, Kim BW, Choi DK (2012) Nuclear factor erythroid 2-related factor 2 signaling in Parkinson disease: a promising multi therapeutic target against oxidative stress, neuroinflammation and cell death. CNS Neurol Disord Drug Targets 11:1015–1029CrossRefPubMedGoogle Scholar
  15. 15.
    Tufekci KU, Civi Bayin E, Genc S, Genc K (2011) The Nrf2/ARE pathway: a promising target to counteract mitochondrial dysfunction in Parkinson’s disease. Parkinsons Dis 2011:314082PubMedCentralPubMedGoogle Scholar
  16. 16.
    Na HK, Kim EH, Jung JH, Lee HH, Hyun JW, Surh YJ (2008) (−)-Epigallocatechin gallate induces Nrf2-mediated antioxidant enzyme expression via activation of PI3K and ERK in human mammary epithelial cells. Arch Biochem Biophys 476:171–177CrossRefPubMedGoogle Scholar
  17. 17.
    Nakaso K, Yano H, Fukuhara Y, Takeshima T, Wada-Isoe K, Nakashima K (2003) PI3K is a key molecule in the Nrf2-mediated regulation of antioxidative proteins by hemin in human neuroblastoma cells. FEBS Lett 546:181–184CrossRefPubMedGoogle Scholar
  18. 18.
    Dai J, Mumper RJ (2010) Plant phenolics: extraction, analysis and their antioxidant and anticancer properties. Molecules 15:7313–7352CrossRefPubMedGoogle Scholar
  19. 19.
    Leem E, Nam JH, Jeon MT, Shin WH, Won SY, Park SJ, Choi MS, Jin BK, Jung UJ, Kim SR (2014) Naringin protects the nigrostriatal dopaminergic projection through induction of GDNF in a neurotoxin model of Parkinson’s disease. J Nutr Biochem 25:801–806CrossRefPubMedGoogle Scholar
  20. 20.
    Qualls Z, Brown D, Ramlochansingh C, Hurley LL, Tizabi Y (2014) Protective effects of curcumin against rotenone and salsolinol-induced toxicity: implications for Parkinson’s disease. Neurotox Res 25:81–89PubMedCentralCrossRefPubMedGoogle Scholar
  21. 21.
    Golechha M, Sarangal V, Bhatia J, Chaudhry U, Saluja D, Arya DS (2014) Naringin ameliorates pentylenetetrazol-induced seizures and associated oxidative stress, inflammation, and cognitive impairment in rats: possible mechanisms of neuroprotection. Epilepsy Behav 41:98–102CrossRefPubMedGoogle Scholar
  22. 22.
    Gopinath K, Prakash D, Sudhandiran G (2011) Neuroprotective effect of naringin, a dietary flavonoid against 3-nitropropionic acid-induced neuronal apoptosis. Neurochem Int 59:1066–1073CrossRefPubMedGoogle Scholar
  23. 23.
    Chen Y, Nie YC, Luo YL, Lin F, Zheng YF, Cheng GH, Wu H, Zhang KJ, Su WW, Shen JG, Li PB (2013) Protective effects of naringin against paraquat-induced acute lung injury and pulmonary fibrosis in mice. Food Chem Toxicol 58:133–140CrossRefPubMedGoogle Scholar
  24. 24.
    Camargo CA, Gomes-Marcondes MC, Wutzki NC, Aoyama H (2012) Naringin inhibits tumor growth and reduces interleukin-6 and tumor necrosis factor α levels in rats with Walker 256 carcinosarcoma. Anticancer Res 32:129–133PubMedGoogle Scholar
  25. 25.
    Chanet A, Milenkovic D, Deval C, Potier M, Constans J, Mazur A, Bennetau-Pelissero C, Morand C, Bérard AM (2012) Naringin, the major grapefruit flavonoid, specifically affects atherosclerosis development in diet-induced hypercholesterolemia in mice. J Nutr Biochem 23:469–477CrossRefPubMedGoogle Scholar
  26. 26.
    Gopinath K, Sudhandiran G (2012) Naringin modulates oxidative stress and inflammation in 3-nitropropionic acid-induced neurodegeneration through the activation of nuclear factor-erythroid 2-related factor-2 signalling pathway. Neuroscience 227:134–143CrossRefPubMedGoogle Scholar
  27. 27.
    Kulasekaran G, Ganapasam S (2015) Protective effect of naringin on 3-nitropropionic acid-induced neurodegeneration through the modulation of matrix metalloproteinases and Glial fibrillary acidic protein. Can J Physiol Pharmacol. doi: 10.1139/cjpp-2015-0035 Google Scholar
  28. 28.
    Wu JB, Fong YC, Tsai HY, Chen YF, Tsuzuki M, Tang CH (2008) Naringin-induced bone morphogenetic protein-2 expression via PI3K, Akt, c-Fos/c-Jun and AP-1 pathway in osteoblasts. Eur J Pharmacol 588:333–341CrossRefPubMedGoogle Scholar
  29. 29.
    Yuan J, Lovejoy DB, Richardson DR (2004) Novel di-2-pyridyl-derived iron chelators with marked and selective antitumor activity: in vitro and in vivo assessment. Blood 104:1450–1458CrossRefPubMedGoogle Scholar
  30. 30.
    King J (1965) In: Van D (ed) Lactate dehydrogenase in practical clinical enzymology. London, Nostrand, pp 83–93Google Scholar
  31. 31.
    Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474CrossRefPubMedGoogle Scholar
  32. 32.
    Sinha AK (1972) Colorimetric assay of catalase. Anal Biochem 47:389–394CrossRefPubMedGoogle Scholar
  33. 33.
    Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WG (1973) Selenium: biochemical rote as a component of glutathione peroxidase. Science 179:588–590CrossRefPubMedGoogle Scholar
  34. 34.
    Staal GE, Visser J, Veeger C (1969) Purification and properties of glutathione reductase of human erythrocytes. Biochem Biophys Acta 185:39–48PubMedGoogle Scholar
  35. 35.
    Moron MS, Depierre JW, Mannervik B (1979) Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 582:67–78CrossRefPubMedGoogle Scholar
  36. 36.
    Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358CrossRefPubMedGoogle Scholar
  37. 37.
    Moreadith RW, Fiskum G (1984) Isolation of mitochondria from ascites tumor cells permeabilized with digitonin. Anal Biochem 137:360–367CrossRefPubMedGoogle Scholar
  38. 38.
    Slater EC, Borner WD Jr (1952) The effect of fluoride on the succinic oxidase system. Biochem J 52:185–196PubMedCentralCrossRefPubMedGoogle Scholar
  39. 39.
    Mutisya EM, Bowling AC, Beal MF (1994) Cortical cytochrome oxidase activity is reduced in Alzheimer’s disease. J Neurochem 63:2179–2184CrossRefPubMedGoogle Scholar
  40. 40.
    Mukherjee SB, Das M, Sudhandiran G, Shaha C (2002) Increase in cytosolic Ca2+ levels through the activation of non-selective cation channels induced by oxidative stress causes mitochondrial depolarization leading to apoptosis-like death in Leishmania donovani promastigotes. J Biol Chem 277:24717–24727CrossRefPubMedGoogle Scholar
  41. 41.
    Dey R, Moraes CT (2000) Lack of oxidative phosphorylation and low mitochondrial membrane potential decrease susceptibility to apoptosis and do not modulate the protective effect of Bcl-x(L) in osteosarcoma cells. J Biol Chem 275:7087–7094CrossRefPubMedGoogle Scholar
  42. 42.
    Yao P, Nussler A, Liu L, Hao L, Song F, Schirmeier A, Nussler N (2007) Quercetin protects human hepatocytes from ethanol-derived oxidative stress by inducing heme oxygenase-1 via the MAPK/Nrf2 pathways. J Hepatol 47:253–261CrossRefPubMedGoogle Scholar
  43. 43.
    Mazzone GL, Nistri A (2011) Delayed neuroprotection by riluzole against excitotoxic damage evoked by kainate on rat organotypic spinal cord cultures. Neuroscience 190:318–327CrossRefPubMedGoogle Scholar
  44. 44.
    Farooqui AA, Ong WY, Horrocks LA (2004) Biochemical aspects of neurodegeneration in human brain: involvement of neural membrane phospholipids and phospholipases A2. Neurochem Res 29:1961–1977CrossRefPubMedGoogle Scholar
  45. 45.
    Barnham KJ, Masters CL, Bush AI (2004) Neurodegenerative diseases and oxidative stress. Nat Rev Drug Discov 3:205–214CrossRefPubMedGoogle Scholar
  46. 46.
    Mattson MP (2000) Apoptosis in neurodegenerative disorders. Nat Rev Mol Cell Biol 1:120–129CrossRefPubMedGoogle Scholar
  47. 47.
    Rosenstock TR, Carvalho AC, Jurkiewicz A, Frussa FR, Smaili SS (2004) Mitochondrial calcium, oxidative stress and apoptosis in a neurodegenerative disease model induced by 3-nitropropionic acid. J Neurochem 88:1220–1228CrossRefPubMedGoogle Scholar
  48. 48.
    Stack EC, Ferro JL, Kim J, Del Signore SJ, Goodrich S, Matson S, Hunt BB, Cormier K, Smith K, Matson WR, Ryu H, Ferrante RJ (2008) Therapeutic attenuation of mitochondrial dysfunction and oxidative stress in neurotoxin models of Parkinson’s disease. Biochim Biophys Acta 1782:151–162CrossRefPubMedGoogle Scholar
  49. 49.
    van Muiswinkel FL, Kuiperij HB (2005) The Nrf2–ARE signalling pathway: promising drug target to combat oxidative stress in neurodegenerative disorders. Curr Drug Targets CNS Neurol Disord 4:267–281CrossRefPubMedGoogle Scholar
  50. 50.
    Noer H, Kristensen BW, Noraberg J, Zimmer J, Gramsbergen JB (2002) 3-Nitropropionic acid neurotoxicity in hippocampal slice cultures: developmental and regional vulnerability and dependency on glucose. Exp Neurol 176:237–246CrossRefPubMedGoogle Scholar
  51. 51.
    Túnez I, Tasset I, Pérez-De La Cruz V, Santamaría A (2010) 3-Nitropropionic acid as a tool to study the mechanisms involved in Huntington’s disease: past, present and future. Molecules 15:878–916CrossRefPubMedGoogle Scholar
  52. 52.
    Jeon SM, Bok SH, Jang MK, Lee MK, Nam KT, Park YB, Rhee SJ, Choi MS (2001) Antioxidative activity of naringin and lovastatin in high cholesterol-fed rabbits. Life Sci 69:2855–2866CrossRefPubMedGoogle Scholar
  53. 53.
    Pérez-De La Cruz V, González-Cortés C, Pedraza-Chaverrí J, Maldonado PD, Andrés-Martínez L, Santamaría A (2006) Protective effect of S-allylcysteine on 3-nitropropionic acid-induced lipid peroxidation and mitochondrial dysfunction in rat brain synaptosomes. Brain Res Bull 68:379–383CrossRefPubMedGoogle Scholar
  54. 54.
    Browne SE, Beal MF (2006) Oxidative damage in Huntington’s disease pathogenesis. Antioxid Redox Signal 8:2061–2073CrossRefPubMedGoogle Scholar
  55. 55.
    Liot G, Bossy B, Lubitz S, Kushnareva Y, Sejbuk N, Bossy-Wetzel E (2009) Complex II inhibition by 3-NP causes mitochondrial fragmentation and neuronal cell death via an NMDA- and ROS-dependent pathway. Cell Death Differ 16:899–909PubMedCentralCrossRefPubMedGoogle Scholar
  56. 56.
    Ishige K, Schubert D, Sagara Y (2001) Flavonoids protect neuronal cells from oxidative stress by three distinct mechanisms. Free Radic Biol Med 30:433–446CrossRefPubMedGoogle Scholar
  57. 57.
    Ly JD, Grubb DR, Lawen A (2003) The mitochondrial membrane potential (deltapsi(m)) in apoptosis; an update. Apoptosis 8:115–128CrossRefPubMedGoogle Scholar
  58. 58.
    Ott M, Gogvadze V, Orrenius S, Zhivotovsky B (2007) Mitochondria, oxidative stress and cell death. Apoptosis 12:913–922CrossRefPubMedGoogle Scholar
  59. 59.
    Oliveira JM (2010) Nature and cause of mitochondrial dysfunction in Huntington’s disease: focusing on huntingtin and the striatum. J Neurochem 114:1–12PubMedGoogle Scholar
  60. 60.
    Oliveira JM, Jekabsons MB, Chen S, Lin A, Rego AC, Gonçalves J, Ellerby LM, Nicholls DG (2007) Mitochondrial dysfunction in Huntington’s disease: the bioenergetics of isolated and in situ mitochondria from transgenic mice. J Neurochem 101:241–249CrossRefPubMedGoogle Scholar
  61. 61.
    Nasr P, Gursahani HI, Pang Z, Bondada V, Lee J, Hadley RW, Geddes JW (2003) Influence of cytosolic and mitochondrial Ca2+, ATP, mitochondrial membrane potential, and calpain activity on the mechanism of neuron death induced by 3-nitropropionic acid. Neurochem Int 43:89–99CrossRefPubMedGoogle Scholar
  62. 62.
    Cory S, Adams JM (2002) The Bcl2 family: regulators of the cellular life-or-death switch. Nat Rev Cancer 2:647–656CrossRefPubMedGoogle Scholar
  63. 63.
    Kim HJ, Song JY, Park HJ, Park HK, Yun DH, Chung JH (2009) Naringin protects against rotenone-induced apoptosis in human neuroblastoma SH-SY5Y cells. Korean J Physiol Pharmacol 13:281–285PubMedCentralCrossRefPubMedGoogle Scholar
  64. 64.
    Mandavilli BS, Boldogh I, Van Houten B (2005) 3-Nitropropionic acid-induced hydrogen peroxide, mitochondrial DNA damage, and cell death are attenuated by Bcl-2 overexpression in PC12 cells. Mol Brain Res 133:215–223CrossRefPubMedGoogle Scholar
  65. 65.
    Calkins MJ, Johnson DA, Townsend JA, Vargas MR, Dowell JA, Williamson TP, Kraft AD, Lee JM, Li J, Johnson JA (2009) The Nrf2/ARE pathway as a potential therapeutic target in neurodegenerative disease. Antioxid Redox Signal 11:497–508PubMedCentralCrossRefPubMedGoogle Scholar
  66. 66.
    Johnson JA, Johnson DA, Kraft AD, Calkins MJ, Jakel RJ, Vargas MR, Chen PC (2008) The Nrf2–ARE pathway: an indicator and modulator of oxidative stress in neurodegeneration. Ann N Y Acad Sci 1147:61–69PubMedCentralCrossRefPubMedGoogle Scholar
  67. 67.
    Scapagnini G, Vasto S, Abraham NG, Caruso C, Zella D, Fabio G (2011) Modulation of Nrf2/ARE pathway by food polyphenols: a nutritional neuroprotective strategy for cognitive and neurodegenerative disorders. Mol Neurobiol 44:192–201CrossRefPubMedGoogle Scholar
  68. 68.
    He X, Chen MG, Ma Q (2008) Activation of Nrf2 in defense against cadmium-induced oxidative stress. Chem Res Toxicol 21:1375–1383CrossRefPubMedGoogle Scholar
  69. 69.
    Brunet A, Datta SR, Greenberg ME (2001) Transcription-dependent and -independent control of neuronal survival by the PI3K–Akt signaling pathway. Curr Opin Neurobiol 11:297–305CrossRefPubMedGoogle Scholar
  70. 70.
    Martin D, Rojo AI, Salinas M, Diaz R, Gallardo G, Alam J, De Galarreta CM, Cuadrado A (2004) Regulation of heme oxygenase-1 expression through the phosphatidylinositol 3-kinase/Akt pathway and the Nrf2 transcription factor in response to the antioxidant phytochemical carnosol. J Biol Chem 279:8919–8929CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Cell Biology Laboratory, Department of BiochemistryUniversity of MadrasChennaiIndia

Personalised recommendations