Molecular and Cellular Biochemistry

, Volume 298, Issue 1–2, pp 187–194 | Cite as

Curcumin attenuates glutamate-induced HT22 cell death by suppressing MAP kinase signaling

  • Hyun-Woo Suh
  • Seongman Kang
  • Ki-Sun Kwon
Original paper


Glutamate induces cell death by upsetting the cellular redox homeostasis, termed oxidative glutamate toxicity, in a mouse hippocampal cell line, HT22. Extracellular signal-regulated kinases (ERK) 1/2 are known key players in this process. Here we characterized the roles of both MAP kinases and cell cycle regulators in mediating oxidative glutamate toxicity and the neuroprotective mechanisms of curcumin in HT22 cells. c-Jun N-terminal kinase (JNK) and p38 kinase were activated during the glutamate-induced HT22 cell death, but at a later stage than ERK activation. Treatment with a JNK inhibitor, SP600125, or a p38 kinase inhibitor, SB203580, partly attenuated this cell death. Curcumin, a natural inhibitor of JNK signaling, protected the HT22 cells from glutamate-induced death at nanomolar concentrations more efficiently than SP600125. These doses of curcumin affected neither the level of intracellular glutathione nor the level of reactive oxygen species, but inactivated JNK and p38 significantly. Moreover, curcumin markedly upregulated a cell-cycle inhibitory protein, p21cip1, and downregulated cyclin D1 levels, which might help the cell death prevention. Our results suggest that curcumin has a neuroprotective effect against oxidative glutamate toxicity by inhibiting MAP kinase signaling and influencing cell-cycle regulation.


Neuroprotection Curcumin Glutamate p21cip1 c-Jun N-terminal kinase Cell cycle 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.



We thank Dr. Chae Young Hwang and Dr. Sung Sup Park for helpful discussion, and Sang-Hyun Min and Sun-Woo Yoon for technical support. Grants from KOSEF, KRF, and KRIBB Research Initiative Program supported this work.


  1. 1.
    Coyle JT, Puttfarcken P (1993) Oxidative stress, glutamate, and neurodegenerative disorders. Science 262:689–695PubMedCrossRefGoogle Scholar
  2. 2.
    Choi DW (1988) Glutamate neurotoxicity and diseases of the nervous system. Neuron 1:623–634PubMedCrossRefGoogle Scholar
  3. 3.
    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 2:1547–1558PubMedCrossRefGoogle Scholar
  4. 4.
    Murphy TH, Schnaar RL, Coyle JT (1990) Immature cortical neurons are uniquely sensitive to glutamate toxicity by inhibition of cystine uptake. Faseb J 4:1624–1633PubMedGoogle Scholar
  5. 5.
    Davis JB, Maher P (1994) Protein kinase C activation inhibits glutamate-induced cytotoxicity in a neuronal cell line. Brain Res 652:169–173PubMedCrossRefGoogle Scholar
  6. 6.
    Maher P, Davis JB (1996) The role of monoamine metabolism in oxidative glutamate toxicity. J Neurosci 16:6394–6401PubMedGoogle Scholar
  7. 7.
    Gwag BJ, Lobner D, Koh JY, Wie MB, Choi DW (1995) Blockade of glutamate receptors unmasks neuronal apoptosis after oxygen-glucose deprivation in vitro. Neuroscience 68:615–619PubMedCrossRefGoogle Scholar
  8. 8.
    Newell DW, Barth A, Papermaster V, Malouf AT (1995) Glutamate and non-glutamate receptor mediated toxicity caused by oxygen and glucose deprivation in organotypic hippocampal cultures. J Neurosci 15:7702–7711PubMedGoogle Scholar
  9. 9.
    Schubert D, Piasecki D (2001) Oxidative glutamate toxicity can be a component of the excitotoxicity cascade. J Neurosci 21:7455–7462PubMedGoogle Scholar
  10. 10.
    Krantic S, Mechawar N, Reix S, Quirion R (2005) Molecular basis of programmed cell death involved in neurodegeneration. Trends Neurosci 28:670–676PubMedGoogle Scholar
  11. 11.
    Anderson ME (1985) Determination of glutathione and glutathione disulfide in biological samples. Methods Enzymol 113:548–555PubMedCrossRefGoogle Scholar
  12. 12.
    Hwang CY, Ryu YS, Chung MS, Kim KD, Park SS, Chae SK, Chae HZ, Kwon KS (2004) Thioredoxin modulates activator protein 1 (AP-1) activity and p27Kip1 degradation through direct interaction with Jab1. Oncogene 23:8868–8875PubMedCrossRefGoogle Scholar
  13. 13.
    Maher P (2001) How protein kinase C activation protects nerve cells from oxidative stress-induced cell death. J Neurosci 21:2929–2938PubMedGoogle Scholar
  14. 14.
    Chen YR, Tan TH (1998) Inhibition of the c-Jun N-terminal kinase (JNK) signaling pathway by curcumin. Oncogene 17:173–178PubMedCrossRefGoogle Scholar
  15. 15.
    Maheshwari RK, Singh AK, Gaddipati J, Srimal RC (2006) Multiple biological activities of curcumin:A short review. Life Sci 78:2081–2087PubMedCrossRefGoogle Scholar
  16. 16.
    Subramanian M, Sreejayan, Rao MN, Devasagayam TP, Singh BB (1994) Diminution of singlet oxygen-induced DNA damage by curcumin and related antioxidants. Mutat Res 311:249–255PubMedGoogle Scholar
  17. 17.
    Thiyagarajan M, Sharma SS (2004) Neuroprotective effect of curcumin in middle cerebral artery occlusion induced focal cerebral ischemia in rats. Life Sci 74:969–985PubMedCrossRefGoogle Scholar
  18. 18.
    Li Y, Maher P, Schubert D (1997) Requirement for cGMP in nerve cell death caused by glutathione depletion. J Cell Biol 139:1317–1324PubMedCrossRefGoogle Scholar
  19. 19.
    Ishige K, Schubert D, Sagara Y (2001) Flavonoids protect neuronal cells from oxidative stress by three distinct mechanisms. Free Radic Biol Med 30:433–446PubMedCrossRefGoogle Scholar
  20. 20.
    Joe B, Lokesh BR (1994) Role of capsaicin, curcumin and dietary n-3 fatty acids in lowering the generation of reactive oxygen species in rat peritoneal macrophages. Biochim Biophys Acta 1224:255–263PubMedCrossRefGoogle Scholar
  21. 21.
    Dargusch R, Schubert D (2002) Specificity of resistance to oxidative stress. J Neurochem 81:1394–1400PubMedCrossRefGoogle Scholar
  22. 22.
    Tanos T, Marinissen MJ, Leskow FC, Hochbaum D, Martinetto H, Gutkind JS, Coso OA (2005) Phosphorylation of c-Fos by members of the p38 MAPK family. Role in the AP-1 response to UV light. J Biol Chem 280:18842–18852PubMedCrossRefGoogle Scholar
  23. 23.
    Mielke K, Herdegen T (2000) JNK and p38 stresskinases–degenerative effectors of signal-transduction-cascades in the nervous system. Prog Neurobiol 61:45–60PubMedCrossRefGoogle Scholar
  24. 24.
    Evan GI, Wyllie AH, Gilbert CS, Littlewood TD, Land H, Brooks M, Waters CM, Penn LZ, Hancock DC (1992) Induction of apoptosis in fibroblasts by c-myc protein. Cell 69:119–128PubMedCrossRefGoogle Scholar
  25. 25.
    Sofer-Levi Y, Resnitzky D (1996) Apoptosis induced by ectopic expression of cyclin D1 but not cyclin E. Oncogene 13:2431–2437PubMedGoogle Scholar
  26. 26.
    Choudhuri T, Pal S, Das T, Sa G (2005) Curcumin selectively induces apoptosis in deregulated cyclin D1-expressed cells at G2 phase of cell cycle in a p53-dependent manner. J Biol Chem 280:20059–20068PubMedCrossRefGoogle Scholar
  27. 27.
    Stanciu M, Wang Y, Kentor R, Burke N, Watkins S, Kress G, Reynolds I, Klann E, Angiolieri MR, Johnson JW, DeFranco DB (2000) Persistent activation of ERK contributes to glutamate-induced oxidative toxicity in a neuronal cell line and primary cortical neuron cultures. J Biol Chem 275:12200–12206PubMedCrossRefGoogle Scholar
  28. 28.
    Murray B, Alessandrini A, Cole AJ, Yee AG, Furshpan EJ (1998) Inhibition of the p44/42 MAP kinase pathway protects hippocampal neurons in a cell-culture model of seizure activity. Proc Natl Acad Sci USA 95:11975–11980PubMedCrossRefGoogle Scholar
  29. 29.
    Runden E, Seglen PO, Haug FM, Ottersen OP, Wieloch T, Shamloo M, Laake JH (1998) Regional selective neuronal degeneration after protein phosphatase inhibition in hippocampal slice cultures: evidence for a MAP kinase-dependent mechanism. J Neurosci 18:7296–7305PubMedGoogle Scholar
  30. 30.
    Alessandrini A, Namura S, Moskowitz MA, Bonventre JV (1999) MEK1 protein kinase inhibition protects against damage resulting from focal cerebral ischemia. Proc Natl Acad Sci USA 96:12866–12869PubMedCrossRefGoogle Scholar
  31. 31.
    Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME (1995) Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 270:1326–1331PubMedCrossRefGoogle Scholar
  32. 32.
    Chen RW, Qin ZH, Ren M, Kanai H, Chalecka-Franaszek E, Leeds P, Chuang DM (2003) Regulation of c-Jun N-terminal kinase, p38 kinase and AP-1 DNA binding in cultured brain neurons: roles in glutamate excitotoxicity and lithium neuroprotection. J Neurochem 84:566–575PubMedCrossRefGoogle Scholar
  33. 33.
    Copani A, Condorelli F, Caruso A, Vancheri C, Sala A, Giuffrida Stella AM, Canonico PL, Nicoletti F, Sortino MA (1999) Mitotic signaling by beta-amyloid causes neuronal death. Faseb J 13:2225–2234PubMedGoogle Scholar
  34. 34.
    Wen Y, Yang S, Liu R, Simpkins JW (2005) Cell-cycle regulators are involved in transient cerebral ischemia induced neuronal apoptosis in female rats. FEBS Lett 579:4591–4599PubMedCrossRefGoogle Scholar
  35. 35.
    Yang Y, Mufson EJ, Herrup K (2003) Neuronal cell death is preceded by cell cycle events at all stages of Alzheimer’s disease. J Neurosci 23:2557–2563PubMedGoogle Scholar
  36. 36.
    Guegan C, Levy V, David JP, Ajchenbaum-Cymbalista F, Sola B (1997) c-Jun and cyclin D1 proteins as mediators of neuronal death after a focal ischaemic insult. Neuroreport 8:1003–1007PubMedCrossRefGoogle Scholar
  37. 37.
    Katchanov J, Harms C, Gertz K, Hauck L, Waeber C, Hirt L, Priller J, von Harsdorf R, Bruck W, Hortnagl H, Dirnagl U, Bhide PG, Endres M (2001) Mild cerebral ischemia induces loss of cyclin-dependent kinase inhibitors and activation of cell cycle machinery before delayed neuronal cell death. J Neurosci 21:5045–5053PubMedGoogle Scholar
  38. 38.
    Timsit S, Rivera S, Ouaghi P, Guischard F, Tremblay E, Ben-Ari Y, Khrestchatisky M (1999) Increased cyclin D1 in vulnerable neurons in the hippocampus after ischaemia and epilepsy: a modulator of in vivo programmed cell death? Eur J Neurosci 11:263–278PubMedCrossRefGoogle Scholar
  39. 39.
    Kuan CY, Schloemer AJ, Lu A, Burns KA, Weng WL, Williams MT, Strauss KI, Vorhees CV, Flavell RA, Davis RJ, Sharp FR, Rakic P (2004) Hypoxia-ischemia induces DNA synthesis without cell proliferation in dying neurons in adult rodent brain. J Neurosci 24:10763–10772PubMedCrossRefGoogle Scholar
  40. 40.
    Ghoneim AI, Abdel-Naim AB, Khalifa AE, El-Denshary ES (2002) Protective effects of curcumin against ischaemia/reperfusion insult in rat forebrain. Pharmacol Res 46:273–279PubMedCrossRefGoogle Scholar
  41. 41.
    Wang Q, Sun AY, Simonyi A, Jensen MD, Shelat PB, Rottinghaus GE, MacDonald RS, Miller DK, Lubahn DE, Weisman GA, Sun GY (2005) Neuroprotective mechanisms of curcumin against cerebral ischemia-induced neuronal apoptosis and behavioral deficits. J Neurosci Res 82:138–148PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2006

Authors and Affiliations

  1. 1.Laboratory of Functional ProteomicsKorea Research Institute of Bioscience and BiotechnologyYusongKorea
  2. 2.Graduate School of BiotechnologyKorea UniversitySeoulKorea

Personalised recommendations