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Τίτλος – Title
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Ρόλος των Καλπαϊνών στο Θάνατο PC12 και PC12-NGF Κύτταρων μετά από Διεγερσιμοτοξικότητα Calpaine Mediated Cell Death in Naive and NGF Treated PC12 Cells after Glutamate Induced Exitotoxicity |
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Συγγραφέας – Author
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Χ. Πουρζιτάκη1, Ι. Κλάγκας1, Γ. Κανέλλος 2, Α. Κριτής2 1Εργαστήριο Φαρμακολογίας, 2Εργαστήριο Φυσιολογίας, Ιατρική Σχολή, Αριστοτέλειο Πανεπιστήμιο, Θεσσαλονίκη, Ελλάς C. Pourzitaki1, I. Klagas1, G. Kanellos2, A. Kritis2 1Laboratory of Pharmacology, 2Laboratory of Physiology, Medical School, P.O. Box 54124, Aristotle University, Thessaloniki, Greece |
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Παραπομπή – Citation
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Πουρζιτάκη,Χ., Κλάγκας,Ι., Κανέλλος,Γ., Κριτής,Α. : Ρόλος των Καλπαϊνών στο Θάνατο PC12 και PC12-NGF Κύτταρων μετά από Διεγερσιμοτοξικότητα, Επιθεώρηση Κλιν. Φαρμακολ. Φαρμακοκινητ. 27: 79-82 (2009)
Pourzitaki,C., Klagas,I., Kanellos,G., Kritis,A. : Calpaine Mediated Cell Death in Naive and NGF Treated PC12 Cells after Glutamate Induced Exitotoxicity, Epitheorese Klin. Farmakol. Farmakokinet. 27: 79-82 (2009)
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Ημερομηνία Δημοσιευσης – Publication Date
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8 Μαρτίου 2009 – 2009-03-08
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Γλώσσα Πλήρους Κειμένου –
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Ελληνικά – Greek |
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Λέξεις κλειδιά – Keywords
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Apoptosis, glutamate, toxicity, PC12, NGF, MDL28170
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Λοιποί Όροι – Other Terms
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Κλινική Μελέτη Clinical Trial |
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Περίληψη – Summary
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– Augmented calcium influx can cause biochemical alterations that involve degradative enzymes (proteases and endonucleases) leeding to neurodegeneration by excitotoxicity. PC12 cells (chromaffinergic rat pheochromocytoma cell line) are a useful model for investigating cell death by glutamate induced excitotoxicity in neurons. Upon treatment with nerve growth factor (NGF), PC12 cells culture shift their phenotype to a post-mitotic, differentiated, neurite-bearing NGF-dependent neuron. The purpose of the present study was to investigate the molecular response of central nervous system to glutamate induced excitotoxicity using PC12 cell cultures, after calpain inhibition treatment in different concentrations. These chromaffinergic transformed cells upon treatment with nerve growth factor (NGF), differentiate to a sympathetic phenotype expressing neurites and excitability. PC12-cells were cultured in full medium DMEM at 37° C with 5% CO2. To cause excitotoxicity from glutamate, undifferentiated and NGF-treated PC12 cultures in DMEM with low glucose concentrations have been exposed for 3h to glutamate in two concentrations (0,5 μM and 10 μΜ). We investigated the involvement of proteolysis in cell death induced by glutamate toxicity, using a calpain inhibitor MDL28170. Cell viability was estimated by a colorimetric method using MTT [3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide. According to our results MDL28170 appears to have neuroprotective effects in excitotoxic injury caused by glutamate treatment. However, calpain inhibition is effective only in low glutamate concentrations. Our results indicate that the mechanisms implicated in cell death by glutamate excitotoxicity seem to differentially engage calpain dependent and independent path-ways. |
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Αναφορές – References
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1. Choi, D.W.: Glutamate neurotoxicity and diseases of the nervous system. Trends Neurosci. 11: 465-469 (1988)
2. Saito K.I, Elce J.S., Hamos J.E., Nixon R.A.: (1993) Widespread activation of calcium-activated neutral proteinase (calpain) in the brain in Alzheimer disease: A potential molecular basis for neuronal degeneration. Proc. Natl. Acad. Sci. USA 90: 2628-2632 (1993) 3. Siesjo B.K.: Cell damage in the brain: a speculative synthesis. J. Cereb. Blood Flow Metab. 1: 155-185 (1981) 4. Greene L.A., Aletta J.M., Rukenstein A., Green S.H.: PC12 pheochromocytoma cells: culture, nerve growth factor treatment, and experimental exploitation. Methods Enzymol. 147: 207-216 (1987) 5. Vaux D.L., Korsmeyer S.J.: Cell death in development. Cell 96: 245-254 (1999) 6. Pourzitaki C., Klagas I., Sioga A., Tzimagiorgis G., Kritis A.: Aspartyl-protease inhibition rescues naive and NGF treated PC12 cells from death after oxygen and glucose deprivation. Epitheor. Klin. Farmakol. Farmakokinet. 25: 98-101 (2007) 7. Rami A., Ferger D., Krieglstein J.: Blockade of calpain proteolytic activity rescues neurons from glutamate excitotoxicity. Neurosci. Res. 27: 93-97 (1997) 8. Denizot F., Lang R.: Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J. Immun. Methods 89: 271-277 (1986) 9. Hansen M.B., Nielsen S.E., Berg K.: Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell killing. J. Immunol. Methods 119: 203-210 (1989) 10. Pourzitaki C., Klagas, I., Kaidoglou, A., Tzimagiorgis, G., Kritis, A.: Caspase dependent and independent cell death in naive and NGF treated PC12 cells after glutamate induced exitotoxicity. Epitheor. Klin. Farmakol. Farmakokinet. 25: 94-97 (2007) 11. Pourzitaki C., Kanellos G., Klagas I., Kritis A.: Combined treatment of aspartyl protease inhibitor and nmda antagonist in pc12 cells after glutamate excitotoxicity. Rev. Clin. Pharmacol. Pharmacokinet. 22: 304-307 (2008) |
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Online ISSN 1011-6575
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