Neurochemical Research

, Volume 41, Issue 9, pp 2267–2277 | Cite as

Gartanin Protects Neurons against Glutamate-Induced Cell Death in HT22 Cells: Independence of Nrf-2 but Involvement of HO-1 and AMPK

  • Xiao-yun Gao
  • Sheng-nan Wang
  • Xiao-hong Yang
  • Wen-jian Lan
  • Zi-wei Chen
  • Jing-kao Chen
  • Jian-hui Xie
  • Yi-fan Han
  • Rong-biao Pi
  • Xiao-bo Yang
Original Paper


Oxidative stress mediates the pathogenesis of neurodegenerative disorders. Gartanin, a natural xanthone of mangosteen, possesses multipharmacological activities. Herein, the neuroprotection capacity of gartanin against glutamate-induced damage in HT22 cells and its possible mechanism(s) were investigated for the first time. Glutamate resulted in cell death in a dose-dependent manner and supplementation of 1–10 µM gartanin prevented the detrimental effects of glutamate on cell survival. Additional investigations on the underlying mechanisms suggested that gartanin could effectively reduce glutamate-induced intracellular ROS generation and mitochondrial depolarization. We further found that gartanin induced HO-1 expression independent of nuclear factor erythroid-derived 2-like 2 (Nrf2). Subsequent studies revealed that the inhibitory effects of gartanin on glutamate-induced apoptosis were partially blocked by small interfering RNA-mediated knockdown of HO-1. Finally, the protein expression of phosphorylation of AMP-activated protein kinase (AMPK) and its downstream signal molecules, Sirtuin activator (SIRT1) and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), increased after gartanin treatment. Taken together, these findings suggest gartanin is a potential neuroprotective agent against glutamate-induced oxidative injury partially through increasing Nrf-2-independed HO-1 and AMPK/SIRT1/PGC-1α signaling pathways.


Gartanin Oxidative stress Neuroprotective Nuclear factor erythroid-derived 2-like 2 Heme oxygenase 1 AMP-activated protein kinase 



Parkinson’s disease


Alzheimer’s disease


Reactive oxygen species


Mitochondrial membrane potential


Heme oxygenase 1


Nuclear factor erythroid-derived 2-like 2


AMP-activated protein kinase


H2DCF-DA dichlorodihydrofluorescein diacetate




Rhodamine 123


Small interfering RNA


Microtubule-associated protein light chain 3


Peroxisome proliferator-activated receptor α


Sirtuin activator 1


Peroxisome proliferator-activated receptor-γ coactivator-1α



This study was supported by Guangdong Provincial International Cooperation Project of Science & Technology (No. 2013B051000038), National Natural Science Foundation of China (No. 31371070) and the Fundamental Research Funds for the Central Universities (No. 15ykjc08b) to R. Pi.

Compliance with Ethical Standards

Conflict of interest

All other authors declare no conflict of interest.


  1. 1.
    Lin MT, Beal MF (2006) Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. Nature 443(7113):787–795PubMedCrossRefGoogle Scholar
  2. 2.
    Forman HJ, Fukuto JM, Torres M (2004) Redox signaling: thiol chemistry defines which reactive oxygen and nitrogen species can act as second messengers. Am J Physiol Cell Physiol 287(2):C246–C256PubMedCrossRefGoogle Scholar
  3. 3.
    Radi E, Formichi P, Battisti C, Federico A (2014) Apoptosis and oxidative stress in neurodegenerative diseases. JAD 42(Suppl 3):S125–S152PubMedGoogle Scholar
  4. 4.
    Pita-Almenar JD, Collado MS, Colbert CM, Eskin A (2006) Different mechanisms exist for the plasticity of glutamate reuptake during early long-term potentiation (LTP) and late LTP. J Neurosci 26(41):10461–10471PubMedCrossRefGoogle Scholar
  5. 5.
    Paoletti P (2011) Molecular basis of NMDA receptor functional diversity. Eur J Neurosci 33(8):1351–1365PubMedCrossRefGoogle Scholar
  6. 6.
    Rudy CC, Hunsberger HC, Weitzner DS, Reed MN (2015) The role of the tripartite glutamatergic synapse in the pathophysiology of Alzheimer’s disease. Aging Dis 6(2):131–148PubMedPubMedCentralCrossRefGoogle Scholar
  7. 7.
    Gardoni F, Bellone C (2015) Modulation of the glutamatergic transmission by dopamine: a focus on Parkinson, Huntington and Addiction diseases. Front Cell Neurosci 9:25PubMedPubMedCentralCrossRefGoogle Scholar
  8. 8.
    Kritis AA, Stamoula EG, Paniskaki KA, Vavilis TD (2015) Researching glutamate-induced cytotoxicity in different cell lines: a comparative/collective analysis/study. Front Cell Neurosci 9:91PubMedPubMedCentralCrossRefGoogle Scholar
  9. 9.
    Murphy TH, Miyamoto M, Sastre A, Schnaar RL, Coyle JT (1989) Glutamate toxicity in a neuronal cell line involves inhibition of cystine transport leading to oxidative stress. Neuron 2(6):1547–1558PubMedCrossRefGoogle Scholar
  10. 10.
    Davis JB, Maher P (1994) Protein kinase C activation inhibits glutamate-induced cytotoxicity in a neuronal cell line. Brain Res 652(1):169–173PubMedCrossRefGoogle Scholar
  11. 11.
    Tan S, Wood M, Maher P (1998) Oxidative stress induces a form of programmed cell death with characteristics of both apoptosis and necrosis in neuronal cells. J Neurochem 71(1):95–105PubMedCrossRefGoogle Scholar
  12. 12.
    van Leyen K, Siddiq A, Ratan RR, Lo EH (2005) Proteasome inhibition protects HT22 neuronal cells from oxidative glutamate toxicity. J Neurochem 92(4):824–830PubMedPubMedCentralCrossRefGoogle Scholar
  13. 13.
    Chen J, Chua KW, Chua CC, Yu H, Pei A, Chua BH, Hamdy RC, Xu X, Liu CF (2011) Antioxidant activity of 7,8-dihydroxyflavone provides neuroprotection against glutamate-induced toxicity. Neurosci Lett 499(3):181–185PubMedCrossRefGoogle Scholar
  14. 14.
    Poteet E, Winters A, Yan LJ, Shufelt K, Green KN, Simpkins JW, Wen Y, Yang SH (2012) Neuroprotective actions of methylene blue and its derivatives. PLoS ONE 7(10):e48279PubMedPubMedCentralCrossRefGoogle Scholar
  15. 15.
    Chao XJ, Chen ZW, Liu AM, He XX, Wang SG, Wang YT, Liu PQ, Ramassamy C, Mak SH, Cui W, Kong AN, Yu ZL, Han YF, Pi RB (2014) Effect of tacrine-3-caffeic acid, a novel multifunctional anti-Alzheimer’s dimer, against oxidative-stress-induced cell death in HT22 hippocampal neurons: involvement of Nrf2/HO-1 pathway. CNS Neurosci Ther 20(9):840–850PubMedCrossRefGoogle Scholar
  16. 16.
    Bramanti V, Grasso S, Tomassoni D, Traini E, Raciti G, Viola M, Li Volti G, Campisi A, Amenta F, Avola R (2015) Effect of growth factors and steroid hormones on heme oxygenase and cyclin D1 expression in primary astroglial cell cultures. J Neurosci Res 93(3):521–529PubMedCrossRefGoogle Scholar
  17. 17.
    Shibahara S, Yoshizawa M, Suzuki H, Takeda K, Meguro K, Endo K (1993) Functional analysis of cDNAs for two types of human heme oxygenase and evidence for their separate regulation. J Biochem 113(2):214–218PubMedGoogle Scholar
  18. 18.
    Abraham NG, Lin JH, Schwartzman ML, Levere RD, Shibahara S (1988) The physiological significance of heme oxygenase. Int J Biochem 20(6):543–558PubMedCrossRefGoogle Scholar
  19. 19.
    Li Volti G, Murabito P (2014) Pharmacologic induction of heme oxygenase-1: it is time to take it seriously*. Crit Care Med 42(8):1967–1968PubMedCrossRefGoogle Scholar
  20. 20.
    Kushida T, Li Volti G, Quan S, Goodman A, Abraham NG (2002) Role of human heme oxygenase-1 in attenuating TNF-alpha-mediated inflammation injury in endothelial cells. J Cell Biochem 87(4):377–385PubMedCrossRefGoogle Scholar
  21. 21.
    Bramanti V, Tomassoni D, Grasso S, Bronzi D, Napoli M, Campisi A, Li Volti G, Ientile R, Amenta F, Avola R (2012) Cholinergic precursors modulate the expression of heme oxigenase-1, p21 during astroglial cell proliferation and differentiation in culture. Neurochem Res 37(12):2795–2804PubMedCrossRefGoogle Scholar
  22. 22.
    Tang GH, Chen ZW, Lin TT, Tan M, Gao XY, Bao JM, Cheng ZB, Sun ZH, Huang G, Yin S (2015) Neolignans from Aristolochia fordiana prevent oxidative stress-induced neuronal death through maintaining the Nrf2/HO-1 pathway in HT22 Cells. J Nat Prod 78(8):1894–1903PubMedCrossRefGoogle Scholar
  23. 23.
    Lee DS, Cha BY, Woo JT, Kim YC, Jang JH (2015) Acerogenin A from Acer nikoense maxim prevents oxidative stress-induced neuronal cell death through Nrf2-mediated heme oxygenase-1 expression in mouse hippocampal HT22 cell line. Molecules (Basel, Switzerland) 20(7):12545–12557Google Scholar
  24. 24.
    Park SY, Jin ML, Kim YH, Kim CM, Lee SJ, Park G (2014) Involvement of heme oxygenase-1 in neuroprotection by sanguinarine against glutamate-triggered apoptosis in HT22 neuronal cells. Environ Toxicol Pharmacol 38(3):701–710PubMedCrossRefGoogle Scholar
  25. 25.
    Son Y, Byun SJ, Pae HO (2013) Involvement of heme oxygenase-1 expression in neuroprotection by piceatannol, a natural analog and a metabolite of resveratrol, against glutamate-mediated oxidative injury in HT22 neuronal cells. Amino Acids 45(2):393–401PubMedCrossRefGoogle Scholar
  26. 26.
    Wang R, Yan H, Tang XC (2006) Progress in studies of huperzine A, a natural cholinesterase inhibitor from Chinese herbal medicine. Acta Pharmacol Sin 27(1):1–26PubMedCrossRefGoogle Scholar
  27. 27.
    Wu TY, Chen CP, Jinn TR (2011) Traditional Chinese medicines and Alzheimer’s disease. Taiwan J Obstet Gynecol 50(2):131–135PubMedCrossRefGoogle Scholar
  28. 28.
    Kim HG, Oh MS (2012) Herbal medicines for the prevention and treatment of Alzheimer’s disease. Curr Pharm Des 18(1):57–75PubMedCrossRefGoogle Scholar
  29. 29.
    Liu QY, Wang YT, Lin LG (2015) New insights into the anti-obesity activity of xanthones from Garcinia mangostana. Food Funct 6(2):383–393PubMedCrossRefGoogle Scholar
  30. 30.
    Chin YW, Kinghorn AD (2008) Structural characterization, biological effects, and synthetic studies on xanthones from mangosteen (Garcinia mangostana), a popular botanical dietary supplement. Mini Rev Org Chem 5(4):355–364PubMedPubMedCentralCrossRefGoogle Scholar
  31. 31.
    Cui J, Hu W, Cai Z, Liu Y, Li S, Tao W, Xiang H (2010) New medicinal properties of mangostins: analgesic activity and pharmacological characterization of active ingredients from the fruit hull of Garcinia mangostana L. Pharmacol Biochem Behav 95(2):166–172PubMedCrossRefGoogle Scholar
  32. 32.
    Li G, Thomas S, Johnson JJ (2013) Polyphenols from the mangosteen (Garcinia mangostana) fruit for breast and prostate cancer. Front Pharmacol 4:80PubMedPubMedCentralGoogle Scholar
  33. 33.
    Obolskiy D, Pischel I, Siriwatanametanon N, Heinrich M (2009) Garcinia mangostana L.: a phytochemical and pharmacological review. PTR 23(8):1047–1065PubMedGoogle Scholar
  34. 34.
    Gutierrez-Orozco F, Failla ML (2013) Biological activities and bioavailability of mangosteen xanthones: a critical review of the current evidence. Nutrients 5(8):3163–3183PubMedPubMedCentralCrossRefGoogle Scholar
  35. 35.
    Pedraza-Chaverri J, Cardenas-Rodriguez N, Orozco-Ibarra M, Perez-Rojas JM (2008) Medicinal properties of mangosteen (Garcinia mangostana). Food Chem Toxicol 46(10):3227–3239PubMedCrossRefGoogle Scholar
  36. 36.
    Quan GH, Oh SR, Kim JH, Lee HK, Kinghorn AD, Chin YW (2010) Xanthone constituents of the fruits of Garcinia mangostana with anticomplement activity. PTR 24(10):1575–1577PubMedGoogle Scholar
  37. 37.
    Shan T, Ma Q, Guo K, Liu J, Li W, Wang F, Wu E (2011) Xanthones from mangosteen extracts as natural chemopreventive agents: potential anticancer drugs. Curr Mol Med 11(8):666–677PubMedPubMedCentralCrossRefGoogle Scholar
  38. 38.
    Govindachari TR, Kalyanaraman PS, Muthukumaraswamy N, Pai BR (1971) Xanthones of Garcinia mangostana Linn. Tetrahedron 27(16):3919–3926CrossRefGoogle Scholar
  39. 39.
    Xu Z, Huang L, Chen XH, Zhu XF, Qian XJ, Feng GK, Lan WJ, Li HJ (2014) Cytotoxic prenylated xanthones from the pericarps of Garcinia mangostana. Molecules (Basel, Switzerland) 19(2):1820–1827Google Scholar
  40. 40.
    Liu Z, Antalek M, Nguyen L, Li X, Tian X, Le A, Zi X (2013) The effect of gartanin, a naturally occurring xanthone in mangosteen juice, on the mTOR pathway, autophagy, apoptosis, and the growth of human urinary bladder cancer cell lines. Nutr Cancer 65(Suppl 1):68–77PubMedPubMedCentralCrossRefGoogle Scholar
  41. 41.
    Jung HA, Su BN, Keller WJ, Mehta RG, Kinghorn AD (2006) Antioxidant xanthones from the pericarp of Garcinia mangostana (Mangosteen). J Agric Food Chem 54(6):2077–2082PubMedCrossRefGoogle Scholar
  42. 42.
    Cosentino K, Garcia-Saez AJ (2014) Mitochondrial alterations in apoptosis. Chem Phys Lipids 181:62–75PubMedCrossRefGoogle Scholar
  43. 43.
    Fukui M, Song JH, Choi J, Choi HJ, Zhu BT (2009) Mechanism of glutamate-induced neurotoxicity in HT22 mouse hippocampal cells. Eur J Pharmacol 617(1–3):1–11PubMedCrossRefGoogle Scholar
  44. 44.
    Zhang H, Davies KJ, Forman HJ (2015) Oxidative stress response and Nrf2 signaling in aging. Free Radic Biol Med 88:314–3336PubMedCrossRefGoogle Scholar
  45. 45.
    Aukkanimart R, Boonmars T, Sriraj P, Songsri J, Laummaunwai P, Waraasawapati S, Boonyarat C, Rattanasuwan P, Boonjaraspinyo S (2015) Anthelmintic, anti-inflammatory and antioxidant effects of Garcinia mangostana extract in hamster opisthorchiasis. Exp Parasitol 154:5–13PubMedCrossRefGoogle Scholar
  46. 46.
    Suttirak W, Manurakchinakorn S (2014) In vitro antioxidant properties of mangosteen peel extract. J Food Sci Technol 51(12):3546–3558PubMedCrossRefGoogle Scholar
  47. 47.
    Xie Z, Sintara M, Chang T, Ou B (2015) Functional beverage of Garcinia mangostana (mangosteen) enhances plasma antioxidant capacity in healthy adults. Food Sci Nutr 3(1):32–38PubMedCrossRefGoogle Scholar
  48. 48.
    Cory S, Huang DC, Adams JM (2003) The Bcl-2 family: roles in cell survival and oncogenesis. Oncogene 22(53):8590–8607PubMedCrossRefGoogle Scholar
  49. 49.
    Dargusch R, Piasecki D, Tan S, Liu Y, Schubert D (2001) The role of Bax in glutamate-induced nerve cell death. J Neurochem 76(1):295–301PubMedCrossRefGoogle Scholar
  50. 50.
    Lim JL, Wilhelmus MM, de Vries HE, Drukarch B, Hoozemans JJ, van Horssen J (2014) Antioxidative defense mechanisms controlled by Nrf2: state-of-the-art and clinical perspectives in neurodegenerative diseases. Arch Toxicol 88(10):1773–1786PubMedCrossRefGoogle Scholar
  51. 51.
    Chen J (2014) Heme oxygenase in neuroprotection: from mechanisms to therapeutic implications. Rev Neurosci 25(2):269–280PubMedCrossRefGoogle Scholar
  52. 52.
    Kang J, Jeong MG, Oh S, Jang EJ, Kim HK, Hwang ES (2014) A FoxO1-dependent, but NRF2-independent induction of heme oxygenase-1 during muscle atrophy. FEBS Lett 588(1):79–85PubMedCrossRefGoogle Scholar
  53. 53.
    Kronke G, Kadl A, Ikonomu E, Bluml S, Furnkranz A, Sarembock IJ, Bochkov VN, Exner M, Binder BR, Leitinger N (2007) Expression of heme oxygenase-1 in human vascular cells is regulated by peroxisome proliferator-activated receptors. Arterioscler Thromb Vasc Biol 27(6):1276–1282PubMedCrossRefGoogle Scholar
  54. 54.
    Ronnett GV, Ramamurthy S, Kleman AM, Landree LE, Aja S (2009) AMPK in the brain: its roles in energy balance and neuroprotection. J Neurochem 109(Suppl 1):17–23PubMedPubMedCentralCrossRefGoogle Scholar
  55. 55.
    Feige JN, Auwerx J (2007) Transcriptional coregulators in the control of energy homeostasis. Trends Cell Biol 17(6):292–301PubMedCrossRefGoogle Scholar
  56. 56.
    Jager S, Handschin C, St-Pierre J, Spiegelman BM (2007) AMP-activated protein kinase (AMPK) action in skeletal muscle via direct phosphorylation of PGC-1alpha. Proc Natl Acad Sci USA 104(29):12017–12022PubMedPubMedCentralCrossRefGoogle Scholar
  57. 57.
    Dagon Y, Avraham Y, Magen I, Gertler A, Ben-Hur T, Berry EM (2005) Nutritional status, cognition, and survival: a new role for leptin and AMP kinase. J Biol Chem 280(51):42142–42148PubMedCrossRefGoogle Scholar
  58. 58.
    Zrelli H, Matsuoka M, Kitazaki S, Zarrouk M, Miyazaki H (2011) Hydroxytyrosol reduces intracellular reactive oxygen species levels in vascular endothelial cells by upregulating catalase expression through the AMPK-FOXO3a pathway. Eur J Pharmacol 660(2–3):275–282PubMedCrossRefGoogle Scholar
  59. 59.
    Hardie DG, Ross FA, Hawley SA (2012) AMPK: a nutrient and energy sensor that maintains energy homeostasis. Nat Rev Mol Cell Biol 13(4):251–262PubMedCrossRefGoogle Scholar
  60. 60.
    Amato S, Man HY (2011) Bioenergy sensing in the brain: the role of AMP-activated protein kinase in neuronal metabolism, development and neurological diseases. Cell Cycle 10(20):3452–3460PubMedPubMedCentralCrossRefGoogle Scholar
  61. 61.
    Weisova P, Davila D, Tuffy LP, Ward MW, Concannon CG, Prehn JH (2011) Role of 5′-adenosine monophosphate-activated protein kinase in cell survival and death responses in neurons. Antioxid Redox Signal 14(10):1863–1876PubMedCrossRefGoogle Scholar
  62. 62.
    Salminen A, Kaarniranta K, Haapasalo A, Soininen H, Hiltunen M (2011) AMP-activated protein kinase: a potential player in Alzheimer’s disease. J Neurochem 118(4):460–474PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Xiao-yun Gao
    • 1
  • Sheng-nan Wang
    • 2
    • 3
  • Xiao-hong Yang
    • 2
    • 3
  • Wen-jian Lan
    • 2
  • Zi-wei Chen
    • 2
    • 3
  • Jing-kao Chen
    • 2
    • 3
  • Jian-hui Xie
    • 4
  • Yi-fan Han
    • 3
    • 5
  • Rong-biao Pi
    • 2
    • 3
    • 6
  • Xiao-bo Yang
    • 4
  1. 1.Department of AnesthesiologyGuangdong Provincial Hospital of Traditional Chinese MedicineGuangzhouChina
  2. 2.Department of Pharmacology and Toxicology, School of Pharmaceutical SciencesSun Yat-Sen UniversityGuangzhouChina
  3. 3.International Joint Laboratory (SYSU-PolyU HK) of Novel Anti-Dementia Drugs of GuangdongGuangzhouChina
  4. 4.Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine SyndromeThe Second Affiliated Hospital of Guangzhou University of Chinese MedicineGuangzhouChina
  5. 5.Department of Applied Biology and Chemical Technology, Institute of Modern Chinese MedicineThe Hong Kong Polytechnic UniversityHung Hom, Hong KongChina
  6. 6.Guangdong Province Key Laboratory of Brain Function and Disease, Zhongshan School of MedicineSun Yat-sen UniversityGuangzhouChina

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