Journal of Molecular Neuroscience

, Volume 45, Issue 1, pp 60–67 | Cite as

Inhibition of Endogenous Hydrogen Sulfide Generation is Associated with Homocysteine-Induced Neurotoxicity: Role of ERK1/2 Activation

  • Xiao-Qing Tang
  • Xin-Tian Shen
  • Yi-E Huang
  • Rong-Qian Chen
  • Yan-Kai Ren
  • Heng-Rong Fang
  • Yuan-Yuan Zhuang
  • Chun-Yan Wang


Both elevated homocysteine and decreased hydrogen sulfide (H2S) are observed in the brains of Alzheimer’s disease (AD) patients. Reactive oxygen species (ROS) overproduction contributes to the neurotoxicity of homocysteine; however, H2S is an endogenous antioxidant gas. Therefore, the aim of this study was to investigate whether the imbalance of proportion to this endogenous protective antioxidant gas is involved in homocysteine-caused neurotoxicity. We show that homocysteine inhibits the generation of endogenous H2S and the expression and activity of cystathionine-β-synthetase (CBS), the main enzyme responsible for the generation of H2S in PC12 cells. S-Adenosylmethionine, an activator of CBS, not only prevents homocysteine-induced inhibition of endogenous H2S production but also attenuates homocysteine-triggered cytotoxicity and accumulation of ROS. We find that activation of ERK1/2 occurs in homocysteine-treated PC12 cells and blockade of ERK1/2 with U0126 abolished the homocysteine-induced cytotoxicity and inhibitory effect on endogenous H2S generation. These results indicate that homocysteine neurotoxicity involves reduction of H2S production, which is caused by inhibition of CBS and mediated by activation of ERK1/2. Our study suggests a promising future of H2S-based therapies for neurodegenerative diseases such as AD.


Homocysteine Hydrogen sulfide Cystathionine-β-synthetase ERK1/2 Neurotoxicity 



This study was supported by Natural Science Foundation of China (81071005 and 30770740), Natural Science Foundation of Hunan Province, China (06JJ2074), China Postdoctoral Science Foundation (2005038233), Plan Project for Scientific Research, Department of Science and Technology, Hunan Province (05FJ3039), and the Research Foundation of Education Bureau of Hunan Province (06C700).


  1. Baydas G, Reiter RJ, Akbulut M, Tuzcu M, Tamer S (2005) Melatonin inhibits neural apoptosis induced by homocysteine in hippocampus of rats via inhibition of cytochrome c translocation and caspase-3 activation and by regulating pro- and anti-apoptotic protein levels. Neuroscience 135:879–886PubMedCrossRefGoogle Scholar
  2. Clarke R, Smith AD, Jobst KA, Refsum H, Sutton L, Ueland PM (1998) Folate, vitamin B12, and serum total homocysteine levels in confirmed Alzheimer disease. Arch Neurol 55:1449–1455PubMedCrossRefGoogle Scholar
  3. Duan WG, Shang J, Jiang ZZ, Yao JC, Yun Y, Yan M, Shu B, Lin Q, Yu ZP, Zhang LY (2009) Rho kinase inhibitor Y-27632 down-regulates norepinephrine synthesis and release in PC12 cells. Basic Clin Pharmacol Toxicol 104:434–440PubMedCrossRefGoogle Scholar
  4. Dwyer BE, Raina AK, Perry G, Smith MA (2004) Homocysteine and Alzheimer's disease: a modifiable risk? Free Radic Biol Med 36:1471–1475PubMedCrossRefGoogle Scholar
  5. Eto K, Asada T, Arima K, Makifuchi T, Kimura H (2002) Brain hydrogen sulfide is severely decreased in Alzheimer's disease. Biochem Biophys Res Commun 293:1485–1488PubMedCrossRefGoogle Scholar
  6. Geng B, Chang L, Pan C, Qi Y, Zhao J, Pang Y, Du J, Tang C (2004) Endogenous hydrogen sulfide regulation of myocardial injury induced by isoproterenol. Biochem Biophys Res Commun 318:756–763PubMedCrossRefGoogle Scholar
  7. Greene LA, Tischler AS (1976) Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci USA 73:2424–2428PubMedCrossRefGoogle Scholar
  8. Gu L, Hu X, Xue Z, Yang J, Wan L, Ren Y, Hertz L, Peng L (2010) Potent homocysteine-induced ERK phosphorylation in cultured neurons depends on self-sensitization via system Xc(−). Toxicol Appl Pharmacol 242:209–223PubMedCrossRefGoogle Scholar
  9. Heinecke JW, Rosen H, Suzuki LA, Chait A (1987) The role of sulfur-containing amino acids in superoxide production and modification of low density lipoprotein by arterial smooth muscle cells. J Biol Chem 262:10098–10103PubMedGoogle Scholar
  10. Ho PI, Collins SC, Dhitavat S, Ortiz D, Ashline D, Rogers E, Shea TB (2001) Homocysteine potentiates beta-amyloid neurotoxicity: role of oxidative stress. J Neurochem 78:249–253PubMedCrossRefGoogle Scholar
  11. Ho PI, Ortiz D, Rogers E, Shea TB (2002) Multiple aspects of homocysteine neurotoxicity: glutamate excitotoxicity, kinase hyperactivation and DNA damage. J Neurosci Res 70:694–702PubMedCrossRefGoogle Scholar
  12. Kim JP, Koh JY, Choi DW (1987) l-Homocysteate is a potent neurotoxin on cultured cortical neurons. Brain Res 437:103–110PubMedCrossRefGoogle Scholar
  13. Kimura H (2002) Hydrogen sulfide as a neuromodulator. Mol Neurobiol 26:13–19PubMedCrossRefGoogle Scholar
  14. Kimura Y, Kimura H (2004) Hydrogen sulfide protects neurons from oxidative stress. FASEB J 18:1165–1167PubMedGoogle Scholar
  15. Kruman II, Culmsee C, Chan SL, Kruman Y, Guo Z, Penix L, Mattson MP (2000) Homocysteine elicits a DNA damage response in neurons that promotes apoptosis and hypersensitivity to excitotoxicity. J Neurosci 20:6920–6926PubMedGoogle Scholar
  16. Li R, Kong Y, Ladisch S (1998) Nerve growth factor-induced neurite formation in PC12 cells is independent of endogenous cellular gangliosides. Glycobiology 8:597–603PubMedCrossRefGoogle Scholar
  17. Linnebank M, Lutz H, Jarre E, Vielhaber S, Noelker C, Struys E, Jakobs C, Klockgether T, Evert BO, Kunz WS et al (2006) Binding of copper is a mechanism of homocysteine toxicity leading to COX deficiency and apoptosis in primary neurons, PC12 and SHSY-5Y cells. Neurobiol Dis 23:725–730PubMedCrossRefGoogle Scholar
  18. Lipton SA, Kim WK, Choi YB, Kumar S, D'Emilia DM, Rayudu PV, Arnelle DR, Stamler JS (1997) Neurotoxicity associated with dual actions of homocysteine at the N-methyl-d-aspartate receptor. Proc Natl Acad Sci USA 94:5923–5928PubMedCrossRefGoogle Scholar
  19. Lowicka E, Beltowski J (2007) Hydrogen sulfide (H2S)—the third gas of interest for pharmacologists. Pharmacol Rep 59:4–24PubMedGoogle Scholar
  20. Miller JW (1999) Homocysteine and Alzheimer's disease. Nutr Rev 57:126–129PubMedGoogle Scholar
  21. Mitsuhashi H, Yamashita S, Ikeuchi H, Kuroiwa T, Kaneko Y, Hiromura K, Ueki K, Nojima Y (2005) Oxidative stress-dependent conversion of hydrogen sulfide to sulfite by activated neutrophils. Shock 24:529–534PubMedCrossRefGoogle Scholar
  22. Moore PK, Bhatia M, Moochhala S (2003) Hydrogen sulfide: from the smell of the past to the mediator of the future? Trends Pharmacol Sci 24:609–611PubMedCrossRefGoogle Scholar
  23. Parsons RB, Waring RH, Ramsden DB, Williams AC (1998) In vitro effect of the cysteine metabolites homocysteic acid, homocysteine and cysteic acid upon human neuronal cell lines. Neurotoxicology 19:599–603PubMedGoogle Scholar
  24. Poddar R, Paul S (2009) Homocysteine-NMDA receptor-mediated activation of extracellular signal-regulated kinase leads to neuronal cell death. J Neurochem 110:1095–1106PubMedCrossRefGoogle Scholar
  25. Prudova A, Bauman Z, Braun A, Vitvitsky V, Lu SC, Banerjee R (2006) S-adenosylmethionine stabilizes cystathionine beta-synthase and modulates redox capacity. Proc Natl Acad Sci USA 103:6489–6494PubMedCrossRefGoogle Scholar
  26. Robert K, Pages C, Ledru A, Delabar J, Caboche J, Janel N (2005) Regulation of extracellular signal-regulated kinase by homocysteine in hippocampus. Neuroscience 133:925–935PubMedCrossRefGoogle Scholar
  27. Selhub J (1999) Homocysteine metabolism. Annu Rev Nutr 19:217–246PubMedCrossRefGoogle Scholar
  28. Seshadri S, Beiser A, Selhub J, Jacques PF, Rosenberg IH, D'Agostino RB, Wilson PW, Wolf PA (2002) Plasma homocysteine as a risk factor for dementia and Alzheimer's disease. N Engl J Med 346:476–483PubMedCrossRefGoogle Scholar
  29. Shearman MS, Hawtin SR, Tailor VJ (1995) The intracellular component of cellular 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) reduction is specifically inhibited by beta-amyloid peptides. J Neurochem 65:218–227PubMedCrossRefGoogle Scholar
  30. Szabo C (2007) Hydrogen sulphide and its therapeutic potential. Nat Rev Drug Discov 6:917–935PubMedCrossRefGoogle Scholar
  31. Tang XQ, Fan LL, Li YJ, Shen XT, Zhuan YY, He JQ, Xu JH, and Hu B (2010a) Inhibition of hydrogen sulfide generation contributes to 1-methy-4-phenylpyridinium ion-induced neurotoxicity. Neurotox Res. doi: 10.1007/s12640-010-9180-4
  32. Tang XQ, Shen XT, Huang YE, Ren YK, Chen RQ, Hu B, He JQ, Yin WL, Xu JH, Jiang ZS (2010b) Hydrogen sulfide antagonizes homocysteine-induced neurotoxicity in PC12 cells. Neurosci Res 68:241–249PubMedCrossRefGoogle Scholar
  33. Tang XQ, Yang CT, Chen J, Yin WL, Tian SW, Hu B, Feng JQ, Li YJ (2008) Effect of hydrogen sulphide on beta-amyloid-induced damage in PC12 cells. Clin Exp Pharmacol Physiol 35:180–186PubMedGoogle Scholar
  34. Van Dam F, Van Gool WA (2009) Hyperhomocysteinemia and Alzheimer's disease: a systematic review. Arch Gerontol Geriatr 48:425–430PubMedCrossRefGoogle Scholar
  35. van den Berg M, van der Knaap MS, Boers GH, Stehouwer CD, Rauwerda JA, Valk J (1995) Hyperhomocysteinaemia; with reference to its neuroradiological aspects. Neuroradiology 37:403–411PubMedCrossRefGoogle Scholar
  36. Walrand S, Valeix S, Rodriguez C, Ligot P, Chassagne J, Vasson MP (2003) Flow cytometry study of polymorphonuclear neutrophil oxidative burst: a comparison of three fluorescent probes. Clin Chim Acta 331:103–110PubMedCrossRefGoogle Scholar
  37. Wang R (2002) Two's company, three's a crowd: can H2S be the third endogenous gaseous transmitter? FASEB J 16:1792–1798PubMedCrossRefGoogle Scholar
  38. Wang S, Hu CP, Jiang DJ, Peng J, Zhou Z, Yuan Q, Nie SD, Jiang JL, Li YJ, Huang KL (2009) All-trans retinoic acid inhibits cobalt chloride-induced apoptosis in PC12 cells: role of the dimethylarginine dimethylaminohydrolase/asymmetric dimethylarginine pathway. J Neurosci Res 87:1938–1946PubMedCrossRefGoogle Scholar
  39. White AR, Huang X, Jobling MF, Barrow CJ, Beyreuther K, Masters CL, Bush AI, Cappai R (2001) Homocysteine potentiates copper- and amyloid beta peptide-mediated toxicity in primary neuronal cultures: possible risk factors in the Alzheimer's-type neurodegenerative pathways. J Neurochem 76:1509–1520PubMedCrossRefGoogle Scholar
  40. Whiteman M, Armstrong JS, Chu SH, Jia-Ling S, Wong BS, Cheung NS, Halliwell B, Moore PK (2004) The novel neuromodulator hydrogen sulfide: an endogenous peroxynitrite ‘scavenger’? J Neurochem 90:765–768PubMedCrossRefGoogle Scholar
  41. Whiteman M, Cheung NS, Zhu YZ, Chu SH, Siau JL, Wong BS, Armstrong JS, Moore PK (2005) Hydrogen sulphide: a novel inhibitor of hypochlorous acid-mediated oxidative damage in the brain? Biochem Biophys Res Commun 326:794–798PubMedCrossRefGoogle Scholar
  42. Yoon S, Seger R (2006) The extracellular signal-regulated kinase: multiple substrates regulate diverse cellular functions. Growth Factors 24:21–44PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Xiao-Qing Tang
    • 1
  • Xin-Tian Shen
    • 1
    • 2
  • Yi-E Huang
    • 2
  • Rong-Qian Chen
    • 1
  • Yan-Kai Ren
    • 1
  • Heng-Rong Fang
    • 1
  • Yuan-Yuan Zhuang
    • 1
  • Chun-Yan Wang
    • 1
  1. 1.Department of Physiology, Medical CollegeUniversity of South ChinaHengyangPeople’s Republic of China
  2. 2.Department of PhysiologyHuaihua Medical CollegeHuaihuaPeople’s Republic of China

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