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

Journal of Pharmacological Sciences

Tadashi Nishiya²  Takahiro Katayama¹  Koji Terada²  Hiroshi Asano²  Osamu Sawada²  Shunsuke Fujii²  Takahiro Horinouchi²  Toshihiko Iwanaga³  Masabumi Minami¹  Yosuke Mai²  Soichi Miwa² 

[1] Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Hokkaido University, Japan;Department of Cellular Pharmacology, Graduate School of Medicine, Hokkaido University, Japan;Laboratory of Histology and Cytology, Graduate School of Medicine, Hokkaido University, Japan

关键词: cigarette smoke extract (CSE);    reactive oxygen species (ROS);    NADPH oxidase (NOX);    apoptosis;    protein kinase C (PKC);   

DOI  :  10.1254/jphs.11166FP

学科分类：药学

来源: Nihon Yakuri Gakkai Henshuubu / Japanese Pharmacological Society

【 摘 要 】

References(44)Cited-By(27)Supplementary materials(1)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)-2H-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 N-acetyl-L-cysteine and reduced glutathione. The membrane damage was prevented by catalase and edaravone (a scavenger of •OH) but not by superoxide dismutase, indicating involvement of •OH. In contrast, the CSE-induced cell apoptosis was resistant to edaravone and induced by authentic H2O2 or O2− generated by the xanthine/xanthine oxidase system, indicating involvement of H2O2 or O2− 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.

【 授权许可】

Unknown