Neurochemical Research

, Volume 35, Issue 12, pp 2107–2116 | Cite as

Thiamine and Oxidants Interact to Modify Cellular Calcium Stores

  • Hsueh-Meei Huang
  • Huan-Lian Chen
  • Gary E. Gibson


Diminished thiamine (vitamin B1) dependent processes and oxidative stress accompany Alzheimer’s disease (AD). Thiamine deficiency in animals leads to oxidative stress. These observations suggest that thiamin may act as an antioxidant. The current experiments first tested directly whether thiamin could act as an antioxidant, and then examined the physiological relevance of the antioxidant properties on oxidant sensitive, calcium dependent processes that are altered in AD. The first group of experiments examined whether thiamin could diminish reactive oxygen species (ROS) or reactive nitrogen species (RNS) produced by two very divergent paradigms. Dose response curves determined the concentrations of t-butyl-hydroperoxide (t-BHP) (ROS production) or 3-morpholinosydnonimine ((SIN-1) (RNS production) to induce oxidative stress within cells. Concentrations of thiamine that reduced the RNS in cells did not diminish the ROS. The second group of experiments tested whether thiamine alters oxidant sensitive aspects of calcium regulation including endoplasmic reticulum (ER) calcium stores and capacitative calcium entry (CCE). Thiamin diminished ER calcium considerably, but did not alter CCE. Thiamine did not alter the actions of ROS on ER calcium or CCE. On the other hand, thiamine diminished the effect of RNS on CCE. These data are consistent with thiamine diminishing the actions of the RNS, but not ROS, on physiological targets. Thus, both experimental approaches suggest that thiamine selectively alters RNS. Additional experiments are required to determine whether diminished thiamine availability promotes oxidative stress in AD or whether the oxidative stress in AD brain diminishes thiamine availability to thiamine dependent processes.


Thiamine Oxidative stress Neurodegenerative disease Calcium 



Bombesin-releasable calcium stores


Balanced salt solution


Capacitative calcium entry


Cyclopiazonic acid


Cytosolic free calcium concentration


6-Carboxy-2′,7′-dichlorodihydro-fluorescein diacetate (acetoxymethyl ester)


Diacetate (4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate)


Dinitrogen trioxide


Dulbecco’s modified Eagle’s medium


Endoplasmic reticulum


Fura-2-acetoxymethyl ester






Glutathione disulfide


Glutathione peroxidase


Glutathione reductase


Nitric oxide




Phosphate-buffered saline


Reactive nitrogen species


Reactive oxygen species










Thiamine deficiency


Thiamine thiol


  1. 1.
    Bettendorff L, Wirtzfeld B, Makarchikov AF, Mazzucchelli G, Frederich M et al (2007) Discovery of a natural thiamine adenine nucleotide. Nat Chem Biol 3:211–212CrossRefPubMedGoogle Scholar
  2. 2.
    Meador K, Loring D, Nichols M, Zamrini E, Rivner M et al (1993) Preliminary findings of high-dose thiamine in dementia of Alzheimer’s type. J Geriatr Psychiatry Neurol 6:222–229PubMedGoogle Scholar
  3. 3.
    Gibson GE, Sheu KF, Blass JP, Baker A, Carlson KC et al (1988) Reduced activities of thiamine-dependent enzymes in the brains and peripheral tissues of patients with Alzheimer’s disease. Arch Neurol 45:836–840PubMedGoogle Scholar
  4. 4.
    Sheu KF, Calingasan NY, Lindsay JG, Gibson GE (1998) Immunochemical characterization of the deficiency of the alpha-ketoglutarate dehydrogenase complex in thiamine-deficient rat brain. J Neurochem 70:1143–1150CrossRefPubMedGoogle Scholar
  5. 5.
    Calingasan NY, Gibson GE (2000) Vascular endothelium is a site of free radical production and inflammation in areas of neuronal loss in thiamine-deficient brain. Ann N Y Acad Sci 903:353–356CrossRefPubMedGoogle Scholar
  6. 6.
    Gibson GE, Zhang H (2002) Interactions of oxidative stress with thiamine homeostasis promote neurodegeneration. Neurochem Int 40:493–504CrossRefPubMedGoogle Scholar
  7. 7.
    Langlais PJ, Anderson G, Guo SX, Bondy SC (1997) Increased cerebral free radical production during thiamine deficiency. Metab Brain Dis 12:137–143PubMedGoogle Scholar
  8. 8.
    Pannunzio P, Hazell AS, Pannunzio M, Rao KV, Butterworth RF (2000) Thiamine deficiency results in metabolic acidosis and energy failure in cerebellar granule cells: an in vitro model for the study of cell death mechanisms in Wernicke’s encephalopathy. J Neurosci Res 62:286–292CrossRefPubMedGoogle Scholar
  9. 9.
    Tolstykh OI, Khmelevskii IuV (1991) The role of alpha-tocopherol and thiamine in the correction of lipid peroxidation in compensatory myocardial hypertrophy. Vopr Pitan 3:38–42Google Scholar
  10. 10.
    Ding Q, Dimayuga E, Keller JN (2007) Oxidative damage, protein synthesis, and protein degradation in Alzheimer’s disease. Curr Alzheimer Res 4:73–79CrossRefPubMedGoogle Scholar
  11. 11.
    Gibson GE, Sheu KF, Blass JP (1998) Abnormalities of mitochondrial enzymes in Alzheimer disease. J Neural Transm 105:855–870CrossRefPubMedGoogle Scholar
  12. 12.
    Mastrogiacoma F, Bettendorff L, Grisar T, Kish SJ (1996) Brain thiamine, its phosphate esters, and its metabolizing enzymes in Alzheimer’s disease. Ann Neurol 39:585–591CrossRefPubMedGoogle Scholar
  13. 13.
    Thyagarajan B, Malli R, Schmidt K, Graier WF, Groschner K (2002) Nitric oxide inhibits capacitative Ca2+ entry by suppression of mitochondrial Ca2+ handling. Br J Pharmacol 137:821–830CrossRefPubMedGoogle Scholar
  14. 14.
    Huang HM, Chen HL, Xu H, Gibson GE (2005) Modification of endoplasmic reticulum Ca2+ stores by select oxidants produces changes reminiscent of those in cells from patients with Alzheimer disease. Free Radic Biol Med 39:979–989CrossRefPubMedGoogle Scholar
  15. 15.
    Huang HM, Zhang H, Ou HC, Chen HL, Gibson GE (2004) Alpha-keto-beta-methyl-n-valeric acid diminishes reactive oxygen species and alters endoplasmic reticulum Ca(2+) stores. Free Radic Biol Med 37:1779–1789CrossRefPubMedGoogle Scholar
  16. 16.
    Putney JW Jr, Ribeiro CM (2000) Signaling pathways between the plasma membrane and endoplasmic reticulum calcium stores. Cell Mol Life Sci 57:1272–1286CrossRefPubMedGoogle Scholar
  17. 17.
    Gibson G, Tofel-Grehl B, Scheffold K, Cristofalo V, Blass J (1998) A reproducible procedure for primary culture and subsequent maintenance of multiple lines of human skin fibroblasts. Age 21Google Scholar
  18. 18.
    Barton D, Le Gloahec V, Patin H, Launay F (1998) Radical chemistry of tert-butyl hydroperoxide (tBHP). Part 1. Studies of the Fe3+-tBHP mechanism. New J Chem 22:559–563Google Scholar
  19. 19.
    Lin WL, Wang CJ, Tsai YY, Liu CL, Hwang JM et al (2000) Inhibitory effect of esculetin on oxidative damage induced by t-butyl hydroperoxide in rat liver. Arch Toxicol 74:467–472CrossRefPubMedGoogle Scholar
  20. 20.
    Ochi T, Miyaura S (1989) Cytotoxicity of an organic hydroperoxide and cellular antioxidant defense system against hydroperoxides in cultured mammalian cells. Toxicology 55:69–82CrossRefPubMedGoogle Scholar
  21. 21.
    Bird GS, Burgess GM, Putney JW Jr (1993) Sulfhydryl reagents and cAMP-dependent kinase increase the sensitivity of the inositol 1, 4, 5-trisphosphate receptor in hepatocytes. J Biol Chem 268:17917–17923PubMedGoogle Scholar
  22. 22.
    Malli R, Frieden M, Trenker M, Graier WF (2005) The role of mitochondria for Ca2+ refilling of the endoplasmic reticulum. J Biol Chem 280:12114–12122CrossRefPubMedGoogle Scholar
  23. 23.
    Poirier SN, Poitras M, Laflamme K, Guillemette G (2001) Thiol-reactive agents biphasically regulate inositol 1, 4, 5-trisphosphate binding and Ca(2 +) release activities in bovine adrenal cortex microsomes. Endocrinology 142:2614–2621CrossRefPubMedGoogle Scholar
  24. 24.
    Anzai K, Ogawa K, Kuniyasu A, Ozawa T, Yamamoto H et al (1998) Effects of hydroxyl radical and sulfhydryl reagents on the open probability of the purified cardiac ryanodine receptor channel incorporated into planar lipid bilayers. Biochem Biophys Res Commun 249:938–942CrossRefPubMedGoogle Scholar
  25. 25.
    Beckman JS, Koppenol WH (1996) Nitric oxide, superoxide, and peroxynitrite: the good, the bad, and ugly. Am J Physiol 271:C1424–C1437PubMedGoogle Scholar
  26. 26.
    Jourd’heuil D, Jourd’heuil FL, Feelisch M (2003) Oxidation and nitrosation of thiols at low micromolar exposure to nitric oxide. Evidence for a free radical mechanism. J Biol Chem 278:15720–15726CrossRefPubMedGoogle Scholar
  27. 27.
    Kharitonov VG, Sundquist AR, Sharma VS (1995) Kinetics of nitrosation of thiols by nitric oxide in the presence of oxygen. J Biol Chem 270:28158–28164CrossRefPubMedGoogle Scholar
  28. 28.
    Kirsch M, Fuchs A, de Groot H (2003) Regiospecific nitrosation of N-terminal-blocked tryptophan derivatives by N2O3 at physiological pH. J Biol Chem 278:11931–11936CrossRefPubMedGoogle Scholar
  29. 29.
    Stepuro AI, Piletskaya TP, Stepuro II (2005) Role of thiamine thiol form in nitric oxide metabolism. Biochemistry (Mosc) 70:339–349CrossRefGoogle Scholar
  30. 30.
    Bischof G, Brenman J, Bredt DS, Machen TE (1995) Possible regulation of capacitative Ca2+ entry into colonic epithelial cells by NO and cGMP. Cell Calcium 17:250–262CrossRefPubMedGoogle Scholar
  31. 31.
    Trepakova ES, Cohen RA, Bolotina VM (1999) Nitric oxide inhibits capacitative cation influx in human platelets by promoting sarcoplasmic/endoplasmic reticulum Ca2+-ATPase-dependent refilling of Ca2+ stores. Circ Res 84:201–209PubMedGoogle Scholar
  32. 32.
    Cohen RA, Weisbrod RM, Gericke M, Yaghoubi M, Bierl C et al (1999) Mechanism of nitric oxide-induced vasodilatation: refilling of intracellular stores by sarcoplasmic reticulum Ca2+ ATPase and inhibition of store-operated Ca2+ influx. Circ Res 84:210–219PubMedGoogle Scholar
  33. 33.
    Doutheil J, Althausen S, Treiman M, Paschen W (2000) Effect of nitric oxide on endoplasmic reticulum calcium homeostasis, protein synthesis and energy metabolism. Cell Calcium 27:107–115CrossRefPubMedGoogle Scholar
  34. 34.
    Viner RI, Williams TD, Schoneich C (1999) Peroxynitrite modification of protein thiols: oxidation, nitrosylation, and S-glutathiolation of functionally important cysteine residue(s) in the sarcoplasmic reticulum Ca-ATPase. Biochemistry 38:12408–12415CrossRefPubMedGoogle Scholar
  35. 35.
    Xu L, Eu JP, Meissner G, Stamler JS (1998) Activation of the cardiac calcium release channel (ryanodine receptor) by poly-S-nitrosylation. Science 279:234–237CrossRefPubMedGoogle Scholar
  36. 36.
    Okai Y, Higashi-Okai K, FS E, Konaka R, Inoue M (2007) Potent radical-scavenging activities of thiamin and thiamin diphosphate. J Clin Biochem Nutr 40:42–48CrossRefPubMedGoogle Scholar
  37. 37.
    Batifoulier F, Verny MA, Besson C, Demigné C, Rémésy C (2005) Determination of thiamine and its phosphate esters in rat tissues analyzed as thiochromes on a RP-amide C16 column. J Chromatogr B 816:67–72CrossRefGoogle Scholar
  38. 38.
    He J, Kang H, Yan F, Chen C (2004) The endoplasmic reticulum-related events in S-nitrosoglutathione-induced neurotoxicity in cerebellar granule cells. Brain Res 1015:25–33CrossRefPubMedGoogle Scholar
  39. 39.
    Calingasan NY, Huang PL, Chun HS, Fabian A, Gibson GE (2000) Vascular factors are critical in selective neuronal loss in an animal model of impaired oxidative metabolism. J Neuropathol Exp Neurol 59:207–217PubMedGoogle Scholar
  40. 40.
    Galdhar NR, Pawar SS (1976) Hepatic drug metabolism and lipid peroxidation in thiamine deficient rats. Int J Vitam Nutr Res 46:14–23PubMedGoogle Scholar
  41. 41.
    Kruse M, Navarro D, Desjardins P, Butterworth RF (2004) Increased brain endothelial nitric oxide synthase expression in thiamine deficiency: relationship to selective vulnerability. Neurochem Int 45:49–56CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Hsueh-Meei Huang
    • 1
  • Huan-Lian Chen
    • 1
  • Gary E. Gibson
    • 1
  1. 1.Burke Medical Research InstituteWeill Medical College of Cornell UniversityWhite PlainsUSA

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