Biopolym. Cell. 2006; 22(4):290-298.
http://dx.doi.org/10.7124/bc.00073B
Cell Biology
Apoptosis activation in rat astrocyte cells caused by antagonists of thiamine
1Chornyy S. A., 1Parkhomenko Yu. M.
  1. Palladin Institute of Biochemistry, NAS of Ukraine
    9, Leontovycha Str., Kyiv, Ukraine, 01601

Abstract

The experimental results of in vitro investigation of rat astrocytes (DITNC cells) reaction to the addition of well-known thiamine (vitamin B1) antagonists into the incubation medium are represented in the work. The following antagonists of thiamine - amprolium, pyrithiamine, and oxythiamine were used. Confocal laser scanning fluorescent microscopy has revealed that 72 hours incubation of cells with oxythiamine or pyrithiamine results in cell membrane vacuolization, externalization of membrane phospholipids and translocation of cleaved caspase 3 to the nucleus. Cleavage of caspase 3 in the cells that incubated with 100 µM concentration of oxythiamine and pyrithiamine was determined by Western blot and imunofluorescence methods. These data prove that incubation of DITNC cells with oxythiamine and pyrithiamine caused the activation of apoptosis processes. At the same conditions amprolium did not influence DITNC cells significantly.
Keywords: thiamine antagonists, oxythiamine, pyrithiamine, amorolium, apoptosis
Full text: (PDF, in Ukrainian)

References

[1] Frederikse PH, Farnsworth P, Zigler JS Jr. Thiamine deficiency in vivo produces fiber cell degeneration in mouse lenses. Biochem Biophys Res Commun. 1999;258(3):703-7.
[2] Stagg AR, Fleming JC, Baker MA, Sakamoto M, Cohen N, Neufeld EJ. Defective high-affinity thiamine transporter leads to cell death in thiamine-responsive megaloblastic anemia syndrome fibroblasts. J Clin Invest. 1999;103(5):723-9.
[3] Park LC, Calingasan NY, Uchida K, Zhang H, Gibson GE. Metabolic impairment elicits brain cell type-selective changes in oxidative stress and cell death in culture. J Neurochem. 2000;74(1):114-24.
[4] Pannunzio P, Hazell AS, Pannunzio M, Rao KV, Butterworth RF. 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. 2000; 62(2): 286-92.
[5] Calingasan NY, Gibson GE. Dietary restriction attenuates the neuronal loss, induction of heme oxygenase-1 and blood-brain barrier breakdown induced by impaired oxidative metabolism. Brain Res. 2000;885(1):62-9.
[6] Kruse M, Navarro D, Desjardins P, Butterworth RF. Increased brain endothelial nitric oxide synthase expression in thiamine deficiency: relationship to selective vulnerability. Neurochem Int. 2004;45(1):49-56.
[7] Park LC, Calingasan NY, Uchida K, Zhang H, Gibson GE. Metabolic impairment elicits brain cell type-selective changes in oxidative stress and cell death in culture. J Neurochem. 2000;74(1):114-24.
[8] Ra?s B, Comin B, Puigjaner J, Brandes JL, Creppy E, Saboureau D, Ennamany R, Lee WN, Boros LG, Cascante M. Oxythiamine and dehydroepiandrosterone induce a G1 phase cycle arrest in Ehrlich's tumor cells through inhibition of the pentose cycle. FEBS Lett. 1999;456(1):113-8.
[9] Wang JJ, Hua Z, Fentress HM, Singleton CK. JNK1 is inactivated during thiamine deficiency-induced apoptosis in human neuroblastoma cells. J Nutr Biochem. 2000;11(4):208-15.
[10] Ostrovsky Y. On the mechanisms of coenzymic and noncoenzymic action of thiamine. J Vitaminol (Kyoto). 1968;14:Suppl:98-102.
[11] Parkhomenko IuM, Donchenko GV, Protasova ZS. [The neural activity of thiamine: facts and hypotheses]. Ukr Biokhim Zh. 1996;68(2):3-14.
[12] Parkhomenko IuM, Chernysh IIu, Churilova TIa, Khalmuradov AG. [Effect of thiamine phosphates on the activity of regulatory enzymes of the pyruvate dehydrogenase complex]. Ukr Biokhim Zh. 1987;59(5):49-54.
[13] Nghi?m HO, Bettendorff L, Changeux JP. Specific phosphorylation of Torpedo 43K rapsyn by endogenous kinase(s) with thiamine triphosphate as the phosphate donor. FASEB J. 2000;14(3):543-54.
[14] Meador K, Loring D, Nichols M, Zamrini E, Rivner M, Posas H, Thompson E, Moore E. Preliminary findings of high-dose thiamine in dementia of Alzheimer's type. J Geriatr Psychiatry Neurol. 1993;6(4):222-9.
[15] H?roux M, Raghavendra Rao VL, Lavoie J, Richardson JS, Butterworth RF. Alterations of thiamine phosphorylation and of thiamine-dependent enzymes in Alzheimer's disease. Metab Brain Dis. 1996;11(1):81-8.
[16] Butterworth RF. Effects of thiamine deficiency on brain metabolism: implications for the pathogenesis of the Wernicke-Korsakoff syndrome. Alcohol Alcohol. 1989;24(4):271-9.
[17] Hazell AS, Todd KG, Butterworth RF. Mechanisms of neuronal cell death in Wernicke's encephalopathy. Metab Brain Dis. 1998;13(2):97-122.
[18] Tong W, Sun GY. Phosphorylation of lipids in rat primary glial cells and immortalized astrocytes (DITNC). Lipids. 1994;29(6):385-90.
[19] R?cker-Martin C, H?naff M, Hatem SN, Delpy E, Mercadier JJ. Early redistribution of plasma membrane phosphatidylserine during apoptosis of adult rat ventricular myocytes in vitro. Basic Res Cardiol. 1999;94(3):171-9.
[20] Godard T, Deslandes E, Lebailly P, Vigreux C, Sichel F, Poul J-M, et al. Early detection of staurosporine-induced apoptosis by comet and annexin V assays. Histochem Cell Biol. 1999;112(2):155–61.
[21] Nagarajah NS, Vigneswaran N, Zacharias W. Hypoxia-mediated apoptosis in oral carcinoma cells occurs via two independent pathways. Mol Cancer. 2004;3(1):38.
[22] Zhang GS, Zhou GB, Dai CW. Upregulation and activation of caspase-3 or caspase-8 and elevation of intracellular free calcium mediated apoptosis of indomethacin-induced K562 cells. Chin Med J (Engl). 2004;117(7):978-84.
[23] Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-54.
[24] Ostrovskiy YuM. Thiamin. Minsk, 1971; 144 p.