Nicotine- and Tar-Free Cigarette Smoke Induces Cell Damage Through Reactive Oxygen Species Newly Generated by PKC-Dependent Activation of NADPH Oxidase

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Author(s)

    • SAWADA Osamu
    • Department of Cellular Pharmacology, Graduate School of Medicine, Hokkaido University
    • FUJII Shunsuke
    • Department of Cellular Pharmacology, Graduate School of Medicine, Hokkaido University
    • NISHIYA Tadashi
    • Department of Cellular Pharmacology, Graduate School of Medicine, Hokkaido University
    • MINAMI Masabumi
    • Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Hokkaido University
    • KATAYAMA Takahiro
    • Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Hokkaido University
    • IWANAGA Toshihiko
    • Laboratory of Histology and Cytology, Graduate School of Medicine, Hokkaido University
    • TERADA Koji
    • Department of Cellular Pharmacology, Graduate School of Medicine, Hokkaido University
    • MIWA Soichi
    • Department of Cellular Pharmacology, Graduate School of Medicine, Hokkaido University

Abstract

We examined cytotoxic effects of nicotine/tar-free cigarette smoke extract (CSE) on C6 glioma cells. The CSE induced plasma membrane damage (determined by lactate dehydrogenase leakage and propidium iodide uptake) and cell apoptosis {determined by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2<I>H</I>-tetrazolium] reduction activity and DNA fragmentation}. The cytotoxic activity decayed with a half-life of approximately 2 h at 37°C, and it was abolished by <I>N</I>-acetyl-L-cysteine and reduced glutathione. The membrane damage was prevented by catalase and edaravone (a scavenger of <SUP>•</SUP>OH) but not by superoxide dismutase, indicating involvement of <SUP>•</SUP>OH. In contrast, the CSE-induced cell apoptosis was resistant to edaravone and induced by authentic H<SUB>2</SUB>O<SUB>2</SUB> or O<SUB>2</SUB><SUP>−</SUP> generated by the xanthine/xanthine oxidase system, indicating involvement of H<SUB>2</SUB>O<SUB>2</SUB> or O<SUB>2</SUB><SUP>−</SUP> in cell apoptosis. Diphenyleneiodonium [NADPH oxidase (NOX) inhibitor] and bisindolylmaleimide I [BIS I, protein kinase C (PKC) inhibitor] abolished membrane damage, whereas they partially inhibited apoptosis. These results demonstrate that 1) a stable component(s) in the CSE activates PKC, which stimulates NOX to generate reactive oxygen species (ROS), causing membrane damage and apoptosis; 2) different ROS are responsible for membrane damage and apoptosis; and 3) part of the apoptosis is caused by oxidants independently of PKC and NOX.

Journal

  • Journal of Pharmacological Sciences

    Journal of Pharmacological Sciences 118(2), 275-287, 2012-02-20

    The Japanese Pharmacological Society

References:  44

Cited by:  1

Codes

  • NII Article ID (NAID)
    10030455160
  • NII NACSIS-CAT ID (NCID)
    AA11806667
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    13478613
  • NDL Article ID
    023439514
  • NDL Call No.
    Z53-D199
  • Data Source
    CJP  CJPref  NDL  J-STAGE 
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