Neoechinulin A Imparts Resistance to Acute Nitrosative Stress in PC12 Cells: A Potential Link of an Elevated Cellular Reserve Capacity for Pyridine Nucleotide Redox Turnover with Cytoprotection
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概要
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Treatment of PC12 cells with fungus-derived alkaloid neoechinulin A for more than 12 h renders the cells resistant to subsequent superoxide (O2−)/nitric oxide (NO) insults derived from 3-morpholinosydnonimine (SIN-1). However, the underlying mechanism(s) remains largely unclear. To elucidate the mechanism(s), we assessed the specificity of the cytoprotection afforded by neoechinulin A treatment using other cytocidal stressors and also clarified the resulting cellular alterations, focusing on the antioxidant and metabolic enzymes systems. Neoechinulin A treatment for more than 12 h endowed PC12 cells with significant resistance to transient NO toxicity, but not persistent NO toxicity, bolus H2O2 toxicity, or oxidative insult from the redox cycling quinone menadione. Cellular antioxidant system profiling revealed no substantial potentiation of the activity of any antioxidant enzyme in lysate from the neoechinulin A-treated cells excluding glutathione (GSH) content, which was significantly decreased (>50%), resulting in a proportional compromise in the thiol-reducing activity of the intact cells. In addition, no differences were observed in the activity for any nicotinamide adenine dinucleotide (phosphate) reduced form (NAD(P)H)-generating enzyme, steady-state NAD(P)H/nicotinamide adenine dinucleotide (phosphate) oxidized form (NAD(P)+) ratios, or the levels of total NAD(P)H. Nevertheless, the neoechinulin A-treated intact cells exhibited increased NAD(P)H redox turnover when driven by extracellular tetrazolium. The structurally inactive analog preechinulin failed to protect cells against NO toxicity or induce these alterations, suggesting their link with the cytoprotective mechanism. These results suggest that neoechinulin A, despite disabling the GSH defense system, confers cytoprotection against nitrosative stresses by elevating the cellular reserve capacity for NAD(P)H generation, which could offset crippling of energy-supplying systems due to nitrosative stress.
著者
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Takeuchi Toshifumi
Department of Applied Biological Science, Tokyo University of Science
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Akashi Soichiro
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Sci
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Watanabe Nobuo
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Sci
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Shibata Yasushi
Department Of Agricultural Chemistry Faculty Of Agriculture Yamaguchi University
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Sugawara Fumio
Department Of Agricultural Chemistry Faculty Of Agriculture Tohoku University
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Fujieda Satoshi
Department Of Applied Biological Science Tokyo University Of Science
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Kuramochi Kouji
Graduate School Of Life And Environmental Sci. Kyoto Prefectural Univ.
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Arai Takao
Department Of Applied Biological Science Faculty Of Science And Technology Science University Of Tok
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Kobayashi Susumu
Department Of Academic Surgery Chiba University Graduate School Of Medicine
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Konishi Kanako
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science
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Shirai Kyo
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science
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Okada Tatsumi
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science
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Takahashi Muneaki
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science
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Nakagawa Tomoya
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science
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Ogura Yuki
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science
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Takeuchi Toshifumi
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science
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Fujieda Satoshi
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science
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Arai Takao
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science
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Kuramochi Kouji
Graduate School of Life and Environmental Sciences, Kyoto Prefectural University
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