Journal of Molecular Neuroscience

, Volume 56, Issue 3, pp 739–750 | Cite as

Modulation of the Nitrergic Pathway via Activation of PPAR-γ Contributes to the Neuroprotective Effect of Pioglitazone Against Streptozotocin-Induced Memory Dysfunction

  • Atish Prakash
  • Anil Kumar
  • Long Chiau Ming
  • Vasudevan Mani
  • Abu Bakar Abdul Majeed


Alzheimer’s disease (AD) is a neurodegenerative disease characterized by impaired memory function and oxidative damage. NO is a major signaling molecule produced in the central nervous system to modulate neurological activity through modulating nitric oxide synthase. Recently, PPAR-γ agonists have shown neuroprotective effects in neurodegenerative disorders. However, there have been only a few studies identifying mechanisms through which cognitive benefits may be exerted. The present study was designed to investigate the possible nitric oxide mechanism in the protective effect of pioglitazone against streptozotocin (STZ)-induced memory dysfunction. Wistar rats were intracerebroventricularly (ICV) injected with STZ. Then rats were treated with pioglitazone, NO modulators [l-arginine and nitro-l-arginine methyl ester (L-NAME)] for 21 days. Behavioral alterations were assessed in between the study period. Animals were sacrificed immediately after behavioral session, and mito-oxidative parameters, TNF-α, IL-6, and caspase-3 activity were measured. STZ-treated rats showed a memory deficit and significantly increased in mito-oxidative damage and inflammatory mediators and apoptosis in the hippocampus. Chronic treatment of pioglitazone significantly improved memory retention and attenuated mito-oxidative damage parameters, inflammatory markers, and apoptosis in STZ-treated rats. However, l-arginine pretreatment with lower dose of pioglitazone has not produced any protective effect as compared to per se. Furthermore, pretreatment of L-NAME significantly potentiated its protective effect, which indicates the involvement of nitric oxide for activation of PPAR-γ action. These results demonstrate that pioglitazone offers protection against STZ-induced memory dysfunction possibly due to its antioxidant, anti-inflammatory, and anti-apoptotic action mediating nitric oxide pathways and, therefore, could have a therapeutic potential in AD.


Pioglitazone Streptozotocin Nitric oxide Oxidative stress Mitochondria 


  1. Allami N et al (2011) Suppression of nitric oxide synthesis by L-NAME reverses the beneficial effects of pioglitazone on scopolamine-induced memory impairment in mice. Eur J Pharmacol 650:240–248PubMedCrossRefGoogle Scholar
  2. Amor S, Puentes F, Baker D, van der Valk P (2010) Inflammation in neurodegenerative diseases. Immunology 129:154–169PubMedCentralPubMedCrossRefGoogle Scholar
  3. Beckman JS, Chen J, Crow JP, Ye YZ (1994) Reactions of nitric-oxide, superoxide and peroxynitrite with superoxide-dismutase in neurodegeneration. Neural Regen 103:371–380Google Scholar
  4. Berger J, Moller DE (2002) The mechanisms of action of PPARs. Annu Rev Med 53:409–435PubMedCrossRefGoogle Scholar
  5. Berman SB, Hastings TG (1999) Dopamine oxidation alters mitochondrial respiration and induces permeability transition in brain mitochondria: implications for Parkinson’s disease. J Neurochem 73:1127–1137Google Scholar
  6. Butow RA, Avadhani NG (2004) Mitochondrial signaling: the retrograde response. Mol Cell 14:1–15PubMedCrossRefGoogle Scholar
  7. Choi BM, Pae HO, Jang SI, Kim YM, Chung HT (2002) Nitric oxide as a pro-apoptotic as well as anti-apoptotic modulator. J Biochem Mol Biol 35:116–126PubMedCrossRefGoogle Scholar
  8. Collino M et al (2006) Modulation of the oxidative stress and inflammatory response by PPAR-gamma agonists in the hippocampus of rats exposed to cerebral ischemia/reperfusion. Eur J Pharmacol 530:70–80PubMedCrossRefGoogle Scholar
  9. Culman J, Zhao Y, Gohlke P, Herdegen T (2007) PPAR-gamma: therapeutic target for ischemic stroke. Trends Pharmacol Sci 28:244–249PubMedCrossRefGoogle Scholar
  10. Dehmer T, Heneka MT, Sastre M, Dichgans J, Schulz JB (2004) Protection by pioglitazone in the MPTP model of Parkinson’s disease correlates with I kappa B alpha induction and block of NF kappa B and iNOS activation. J Neurochem 88:494–501Google Scholar
  11. Duff K et al (2010) Mild cognitive impairment in prediagnosed Huntington disease. Neurology 75:500–507PubMedCentralPubMedCrossRefGoogle Scholar
  12. Eliasson MJL et al (1999) Neuronal nitric oxide synthase activation and peroxynitrite formation in ischemic stroke linked to neural damage. J Neurosci 19:5910–5918PubMedGoogle Scholar
  13. Ellman GL (1959) Tissue Sulfhydryl Groups. Arch Biochem Biophys 82:70–77Google Scholar
  14. Ellman Gl, Courtney KD, Andres V Jr, Feather-Stone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95Google Scholar
  15. Feil R, Kleppisch T (2008) NO/cGMP-dependent modulation of synaptic transmission. Handb Exp Pharmacol. 184:529–560Google Scholar
  16. Feinstein DL (2003) Therapeutic potential of peroxisome proliferator-activated receptor agonists for neurological disease. Diabetes Technol Ther 5:67–73PubMedCrossRefGoogle Scholar
  17. Feinstein DL et al (2005) Receptor-independent actions of PPAR thiazolidinedione agonists: is mitochondrial function the key? Biochem Pharmacol 70:177–188PubMedCrossRefGoogle Scholar
  18. Fuenzalida K et al (2007) Peroxisome proliferator-activated receptor gamma up-regulates the Bcl-2 anti-apoptotic protein in neurons and induces mitochondrial stabilization and protection against oxidative stress and apoptosis. J Biol Chem 282:37006–37015PubMedCrossRefGoogle Scholar
  19. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR (1982) Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem 126:131–138PubMedCrossRefGoogle Scholar
  20. Grunblatt E, Salkovic-Petrisic M, Osmanovic J, Riederer P, Hoyer S (2007) Brain insulin system dysfunction in streptozotocin intracerebroventricularly treated rats generates hyperphosphorylated tau protein. J Neurochem 101:757–770PubMedCrossRefGoogle Scholar
  21. Guo J et al (2010) Impaired neural stem/progenitor cell proliferation in streptozotocin-induced and spontaneous diabetic mice. Neurosci Res 68:329–336PubMedCrossRefGoogle Scholar
  22. Ha HC, Snyder SH (1999) Poly(ADP-ribose) polymerase is a mediator of necrotic cell death by ATP depletion. Proc Natl Acad Sci U S A 96:13978–13982PubMedCentralPubMedCrossRefGoogle Scholar
  23. Hoyer S, Muller D, Plaschke K (1994) Desensitization of brain insulin receptor. Effect on glucose/energy and related metabolism. J Neural Transm Suppl 44:259–268PubMedGoogle Scholar
  24. Kapadia R, Yi JH, Vemuganti R (2008) Mechanisms of anti-inflammatory and neuroprotective actions of PPAR-gamma agonists. Front Biosci 13:1813–1826PubMedCentralPubMedCrossRefGoogle Scholar
  25. Kaundal RK, Sharma SS (2010) Peroxisome proliferator-activated receptor gamma agonists as neuroprotective agents. Drug News Perspect 23:241–256PubMedCrossRefGoogle Scholar
  26. Kiaei M, Kipiani K, Chen J, Calingasan NY, Beal MF (2005) Peroxisome proliferator-activated receptor-gamma agonist extends survival in transgenic mouse model of amyotrophic lateral sclerosis. Exp Neurol 191:331–336PubMedCrossRefGoogle Scholar
  27. King TE (1967) Preparation of succinate dehydrogenase and reconstitution of succinate oxidase. Methods Enzymol 10:322-331Google Scholar
  28. King TE, Howard RL (1967) Preparations and properties of soluble NADH dehydrogenases from cardiac muscle. Methods Enzymol 10:275-294Google Scholar
  29. Kono Y (1978) Generation of superoxide radical during autoxidation of hydroxylamine and an assay for superoxide dismutase. Arch Biochem Biophys 186:189–195PubMedCrossRefGoogle Scholar
  30. Kubera M, Obuchowicz E, Goehler L, Brzeszcz J, Maes M (2011) In animal models, psychosocial stress-induced (neuro)inflammation, apoptosis and reduced neurogenesis are associated to the onset of depression. Prog Neuropsychopharmacol Biol Psychiatry 35:744–759PubMedCrossRefGoogle Scholar
  31. Kumar A, Vashist A, Kumar P (2010) Potential role of pioglitazone, caffeic acid and their combination against fatigue syndrome-induced behavioural, biochemical and mitochondrial alterations in mice. Inflammopharmacology 18:241–251PubMedCrossRefGoogle Scholar
  32. Landreth GE, Heneka MT (2001) Anti-inflammatory actions of peroxisome proliferator-activated receptor gamma agonists in Alzheimer’s disease. Neurobiol Aging 22:937–944PubMedCrossRefGoogle Scholar
  33. Law A, Gauthier S, Quirion R (2001) Say NO to Alzheimer’s disease: the putative links between nitric oxide and dementia of the Alzheimer’s type. Brain Res Rev 35:73–96PubMedCrossRefGoogle Scholar
  34. Li Z et al (2010) Caspase-3 activation via mitochondria is required for long-term depression and AMPA receptor internalization. Cell 141:859–871PubMedCentralPubMedCrossRefGoogle Scholar
  35. Liu Y, Peterson DA, Kimura H, Schubert D (1997) Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. J Neurochem 69:581–593PubMedCrossRefGoogle Scholar
  36. Liu RL, Liu W, Doctrow SR, Baudry M (2003) Iron toxicity in organotypic cultures of hippocampal slices: role of reactive oxygen species. J Neurochem 85:492–502PubMedCrossRefGoogle Scholar
  37. Liu P, Fleete MS, Jing Y, Collie ND, Curtis MA, Waldvogel HJ, Faull RL, Abraham WC, Zhang H (2014) Altered arginine metabolism in Alzheimer’s disease brains. Neurobiol Aging 35:1992–2003PubMedCrossRefGoogle Scholar
  38. Luo Y, Yue W, Quan X, Wang Y, Zhao B, Lu Z (2015) Asymmetric dimethylarginine exacerbates Aβ-induced toxicity and oxidative stress in human cell and Caenorhabditis elegans models of Alzheimer disease. Free Radic Biol Med 79:117–126PubMedCrossRefGoogle Scholar
  39. Matsumoto T, Noguchi E, Kobayashi T, Kamata K (2007) Mechanisms underlying the chronic pioglitazone treatment-induced improvement in the impaired endothelium-dependent relaxation seen in aortas from diabetic rats. Free Radic Biol Med 42:993–1007PubMedCrossRefGoogle Scholar
  40. Merrill JE, Benveniste EN (1996) Cytokines in inflammatory brain lesions: helpful and harmful. Trends Neurosci 19:331–338PubMedCrossRefGoogle Scholar
  41. Meyer RC, Spangler EL, Patel N, London ED, Ingram DK (1998) Impaired learning in rats in a 14-unit T-maze by 7-nitroindazole, a neuronal nitric oxide synthase inhibitor, is attenuated by the nitric oxide donor, molsidomine. Eur J Pharmacol 341:17–22PubMedCrossRefGoogle Scholar
  42. Miglio G, Rosa AC, Rattazzi L, Collino M, Lombardi G, Fantozzi R (2009) PPAR gamma stimulation promotes mitochondrial biogenesis and prevents glucose deprivation-induced neuronal cell loss. Neurochem Int 55:496–504PubMedCrossRefGoogle Scholar
  43. Moreno S, Farioli-Vecchioli S, Ceru MP (2004) Immunolocalization of peroxisome proliferator-activated receptors and retinoid X receptors in the adult rat CNS. Neuroscience 123:131–145PubMedCrossRefGoogle Scholar
  44. Nicolakakis N, Hamel E (2010) The nuclear receptor PPARgamma as a therapeutic target for cerebrovascular and brain dysfunction in Alzheimer’s disease. Front Aging Neurosci. doi: 10.3389/fnagi.2010.00021 PubMedCentralPubMedGoogle Scholar
  45. Penna C, Perrelli MG, Pagliaro P (2012) Mitochondrial pathways, permeability transition pore and redox signaling in cardioprotection: therapeutic implications. Antioxid Redox Signal 18(5):556–599PubMedCrossRefGoogle Scholar
  46. Pike KE et al (2011) Cognition and beta-amyloid in preclinical Alzheimer’s disease: data from the AIBL study. Neuropsychologia 49:2384–2390PubMedCrossRefGoogle Scholar
  47. Piotrowski P, Wierzbicka K, Smialek M (2001) Neuronal death in the rat hippocampus in experimental diabetes and cerebral ischaemia treated with antioxidants. Folia Neuropathol 39:147–154PubMedGoogle Scholar
  48. Prakash AK, Kumar A (2009) Effect of chronic treatment of carvedilol on oxidative stress in an intracerebroventricular streptozotocin induced model of dementia in rats. J Pharm Pharmacol 61:1665–1672PubMedCrossRefGoogle Scholar
  49. Prakash A, Kumar A (2013) Pioglitazone alleviates the mitochondrial apoptotic pathway and mito-oxidative damage in the d-galactose-induced mouse model. Clin Exp Pharmacol Physiol 40:644–651PubMedCrossRefGoogle Scholar
  50. Prickaerts J, Fahrig T, Blokland A (1999) Cognitive performance and biochemical markers in septum, hippocampus and striatum of rats after an i.c.v. injection of streptozotocin: a correlation analysis. Behav Brain Res 102:73–88Google Scholar
  51. Raza H, John A (2012) Streptozotocin-induced cytotoxicity, oxidative stress and mitochondrial dysfunction in human hepatoma HepG2 cells. Int J Mol Sci 13:5751–5767PubMedCentralPubMedCrossRefGoogle Scholar
  52. Riobo NA et al (2001) Nitric oxide inhibits mitochondrial NADH:ubiquinone reductase activity through peroxynitrite formation. Biochem J 359:139–145PubMedCentralPubMedCrossRefGoogle Scholar
  53. Salkovic-Petrisic M, Hoyer S (2007) Central insulin resistance as a trigger for sporadic Alzheimer-like pathology: an experimental approach. J Neural Transm Suppl 72:217–233Google Scholar
  54. Sauerbeck A et al (2011) Pioglitazone attenuates mitochondrial dysfunction, cognitive impairment, cortical tissue loss, and inflammation following traumatic brain injury. Exp Neurol 227:128–135PubMedCentralPubMedCrossRefGoogle Scholar
  55. Schnegg CI, Robbins ME (2011) Neuroprotective mechanisms of PPARdelta: modulation of oxidative stress and inflammatory processes. PPAR Res 2011:373560PubMedCentralPubMedCrossRefGoogle Scholar
  56. Siemers E (2011) Designing clinical trials for early (pre-dementia) Alzheimer’s disease: determining the appropriate population for treatment. J Nutr Health Aging 15:22–24PubMedCrossRefGoogle Scholar
  57. Sottocasa GL, Kuylenstierna B, Ernster L, Bergstrand A (1967) An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study. J Cell Biol 32:415–438PubMedCentralPubMedCrossRefGoogle Scholar
  58. Steinert JR, Chernova T, Forsythe ID (2010) Nitric oxide signaling in brain function, dysfunction, and dementia. Neuroscientist 16:435–452PubMedCrossRefGoogle Scholar
  59. Storer PD, Xu J, Chavis J, Drew PD (2005) Peroxisome proliferator-activated receptor-gamma agonists inhibit the activation of microglia and astrocytes: implications for multiple sclerosis. J Neuroimmunol 161:113–122PubMedCrossRefGoogle Scholar
  60. Susswein AJ, Katzoff A, Miller N, Hurwitz I (2004) Nitric oxide and memory. Neuroscientist 10:153–162PubMedCrossRefGoogle Scholar
  61. Tan SE (2007) Roles of hippocampal nitric oxide and calcium/calmodulin-dependent protein kinase II in inhibitory avoidance learning in rats. Behav Pharmacol 18:29–38PubMedCrossRefGoogle Scholar
  62. Tota S, Kamat PK, Shukla R, Nath C (2011) Improvement of brain energy metabolism and cholinergic functions contributes to the beneficial effects of silibinin against streptozotocin induced memory impairment. Behav Brain Res 221:207–215PubMedCrossRefGoogle Scholar
  63. Turrens JF (1997) Superoxide production by the mitochondrial respiratory chain. Biosci Rep 17:3–8Google Scholar
  64. Valina LD, Ted MD (1996) Nitric oxide neurotoxicity. J Chem Neuroanat 10:179–190CrossRefGoogle Scholar
  65. Van Dyke K, Jabbour N, Hoeldtke R, Van Dyke C, Van Dyke M (2010) Oxidative/nitrosative stresses trigger type I diabetes: preventable in streptozotocin rats and detectable in human disease. Ann N Y Acad Sci 1203:138–145PubMedCrossRefGoogle Scholar
  66. Wills ED (1966) Mechanisms of lipid peroxide formation in animal tissues. Biochem J 99:667–676PubMedCentralPubMedGoogle Scholar
  67. Yamada K et al (1995) Role of nitric oxide in learning and memory and in monoamine metabolism in the rat brain. Br J Pharmacol 115:852–858PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Atish Prakash
    • 1
    • 2
    • 3
  • Anil Kumar
    • 3
  • Long Chiau Ming
    • 1
    • 2
  • Vasudevan Mani
    • 1
    • 2
  • Abu Bakar Abdul Majeed
    • 1
    • 2
  1. 1.Faculty of Pharmacy, Campus Puncak AlamUniversiti Teknologi MARA (UiTM)Bandar Puncak AlamMalaysia
  2. 2.Brain Degeneration and Therapeutics Group, Brain and Neuroscience Communities of ResearchUniversiti Teknologi MARA (UiTM)Shah AlamMalaysia
  3. 3.Pharmacology Division, University Institute of Pharmaceutical Sciences, UGC Center of Advanced StudyPanjab UniversityChandigarhIndia

Personalised recommendations