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Human Neuroblastoma SH-SY5Y Cells (human + neuroblastoma_sh-sy5y_cell)
Selected AbstractsOxidative modification of mitochondrial proteins and cell death in Parkinson's diseaseJOURNAL OF NEUROCHEMISTRY, Issue 2002W. Maruyama Oxidative stress is one of the cell death mechanisms in neurodegenerative disorders, such as Parkinson's disease (PD) and Alzheimer's disease. Most of reactive oxygen species (ROS) generate in mitochondria through oxidative phosphorylation, and a part of them are not scavenged by antioxidative system and react with bioactive molecules. Recently, alpha-synuclein containing nitrotyrosine, a marker for oxidative modification by peroxynitrite, was identified in Lewy body. In addition, inhibitors of mitochondrial respiratory chain were reported to induce formation of Lewy body-like inclusion in vivo and in vitro. In this paper it was examined whether ROS and reactive nitrogen species (RNS) generated in mitochondria oxidize mitochondrial respiratory enzymes and induce the formation of inclusion body and cell death in PD. Human neuroblastoma SH-SY5Y cells were treated with a peroxynitrite donor, SIN-1, or an inhibitor of complex I, rotenone. After the treatment, proteins modified with toxic aldehydes, 4-hydroxynonenal and acrolein, and containing nitrotyrosine were analyzed by immunoblotting. Particularly in mitochondrial fraction, the oxidized protein was characterized by two-dimensional immunoblotting. Most of the oxidized proteins were detected in subunits proteins of complex I. These results indicate that mitochondrial complex I is a main target of oxidative stress in dopamine neurons and its dysfunction may be involved in the death mechanism in neurodegenerative disorders. [source] Opposing Actions of Phosphatidylinositol 3-Kinase and Glycogen Synthase Kinase-3, in the Regulation of HSF-1 ActivityJOURNAL OF NEUROCHEMISTRY, Issue 6 2000Gautam N. Bijur Abstract: Elevated temperatures activate the survival promoters Aktand heat shock factor-1 (HSF-1), a transcription factor that induces theexpression of heat shock proteins (HSPs), such as HSP-70. Because neuronalmechanisms controlling these responses are not known, these were investigatedin human neuroblastoma SH-SY5Y cells. Heat shock (45°C) rapidly activatedAkt, extracellular signal-regulated kinases 1 and 2 (ERK1/2), and p38, butonly Akt was activated in a phosphatidylinositol 3-kinase (PI-3K)-dependentmanner, as the PI-3K inhibitors LY294002 and wortmannin blocked Aktactivation, but not ERK1/2 or p38 activation. Akt activation was not blockedby inhibition of p38 or ERK1/2, indicating the independence of these signalingsystems. Heat shock treatment also caused a rapid increase in HSF-1 DNAbinding activity that was partially dependent on PI-3K activity, as both thePI-3K inhibitors attenuated this response. Because Akt inhibits glycogensynthase kinase-3, (GSK-3,), an enzyme that facilitates cell death,we tested if GSK-3, is a negative regulator of HSF-1 activation.Overexpression of GSK-3, impaired heat shock-induced activation of HSF-1,and also reduced HSP-70 production, which was partially restored by theGSK-3, inhibitor lithium. Thus, heat shock-induced activation of PI-3Kand the inhibitory effect of GSK-3, on HSF-1 activation and HSP-70expression imply that Akt-induced inhibition of GSK-3, contributes to theactivation of HSF-1. [source] Isoflurane attenuates dynorphin-induced cytotoxicity and downregulation of Bcl-2 expression in differentiated neuroblastoma SH-SY5Y cellsACTA ANAESTHESIOLOGICA SCANDINAVICA, Issue 1 2009G.-J. WU Background: It has been proposed that the volatile anesthetic isoflurane induces neuroprotection and that the endogenous opioid peptide dynorphin induces neurocytotoxicity in cells. The levels of dynorphin are often significantly elevated in neuropathophysiological conditions, and dynorphin can directly induce toxicity. However, the neuroprotective effects of isoflurane on dynorphin-induced cytotoxicity are still unclear. Methods: In order to determine the effect of isoflurane on dynorphin-induced cytotoxicity in neuronal cells, we have designed a device wherein cultured human neuroblastoma SH-SY5Y cells can be exposed to isoflurane. Fully differentiated SH-SY5Y cells were obtained by treating the cells with retinoic acid for 6 days. We examined SH-SY5Y cell survival, apoptosis, and antiapoptotic protein expression by cell viability, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling stain, and Western blot analysis, respectively. Results: After 16 h of dynorphin (10 ,M) treatment, the SH-SY5Y cells showed significant cytotoxicity, apoptosis, and downregulation of the antiapoptotic Bcl-2 protein expression. These effects of dynorphin were significantly inhibited by isoflurane exposure for 32 h [pretreatment for 16 h and posttreatment (after dynorphin treatment) for 16 h]. Conclusion: Thus, our results suggest that isoflurane exerts neuroprotective effects in the case of dynorphin-induced pathophysiological disruption. [source] Effect of chronic Li+ treatment on free intracellular Mg2+ in human neuroblastoma SH-SY5Y cellsBIPOLAR DISORDERS, Issue 1 2003Abde M Abukhdeir Objectives:, Previous findings have demonstrated Li+/Mg2+ competition at therapeutic intracellular Li+ levels after acute Li+ treatment in human neuroblastoma SH-SY5Y cells. In the current study, we examined whether Li+/Mg2+ competition exists at therapeutically relevant extra- and intracellular [Li+] after chronic Li+ loading times. Methods:, In human neuroblastoma cells, intracellular free Mg2+ was determined by fluorescence spectroscopy with the fluorophore furaptra. Intracellular Li+ and Mg2+ were measured by atomic absorption spectrophotometry. Results:, After loading of the neuroblastoma cells with 1,2 mM extracellular Li+ for 24,72 h, the observed, increased intracellular free [Mg2+] levels were significantly higher (p <,0.03) than those in matched Li+ free cells, and intracellular [Li+] was found to be at therapeutic intracellular levels (0.7,1.5 mM). Conclusions:, The results demonstrate that Li+/Mg2+ competition exists after chronic treatment with Li+ at therapeutically relevant intracellular Li+ levels in neuroblastoma cells. We found differences between acute and chronic Li+ treatment effects on the extent of Li+/Mg2+ competition. Possible reasons for these differences are discussed. [source] | ||||||||||