Carbocisteine inhibits oxidant-induced apoptosis in cultured human airway epithelial cells
Corresponding Author
Motoki YOSHIDA
Departments of Geriatrics and Gerontology and
Motoki Yoshida, Department of Geriatrics and Gerontology, Tohoku University School of Medicine, Seiryo-machi, Aoba-ku, Sendai, 980-8574 Japan. Email: [email protected]Search for more papers by this authorKatsutoshi NAKAYAMA
Division of Respiratory Diseases, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, and
Search for more papers by this authorHiroyasu YASUDA
Department of Translational Clinical Oncology, Kyoto University Graduate School of Medicine, Kyoto, Japan
Search for more papers by this authorHiroshi KUBO
Advanced Preventive Medicine for Infectious Disease, Tohoku University School of Medicine, Sendai,
Search for more papers by this authorKazuyoshi KUWANO
Division of Respiratory Diseases, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, and
Search for more papers by this authorMutsuo YAMAYA
Advanced Preventive Medicine for Infectious Disease, Tohoku University School of Medicine, Sendai,
Search for more papers by this authorCorresponding Author
Motoki YOSHIDA
Departments of Geriatrics and Gerontology and
Motoki Yoshida, Department of Geriatrics and Gerontology, Tohoku University School of Medicine, Seiryo-machi, Aoba-ku, Sendai, 980-8574 Japan. Email: [email protected]Search for more papers by this authorKatsutoshi NAKAYAMA
Division of Respiratory Diseases, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, and
Search for more papers by this authorHiroyasu YASUDA
Department of Translational Clinical Oncology, Kyoto University Graduate School of Medicine, Kyoto, Japan
Search for more papers by this authorHiroshi KUBO
Advanced Preventive Medicine for Infectious Disease, Tohoku University School of Medicine, Sendai,
Search for more papers by this authorKazuyoshi KUWANO
Division of Respiratory Diseases, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, and
Search for more papers by this authorMutsuo YAMAYA
Advanced Preventive Medicine for Infectious Disease, Tohoku University School of Medicine, Sendai,
Search for more papers by this authorConflict of interest statement: Matsuo Yamaya and Hiroshi Kubo have received funding from Kyorin Pharmaceutical Co., Ltd. for research carried out as part of this study.
ABSTRACT
Background and objective: Increased oxidant levels have been associated with exacerbations of COPD, and L-carbocisteine, a mucolytic agent, reduces the frequency of exacerbations. The mechanisms underlying the inhibitory effects of L-carbocisteine on oxidant-induced COPD exacerbations were examined in an in vitro study of human airway epithelial cells.
Methods: In order to examine the antioxidant effects of L-carbocisteine, human tracheal epithelial cells were treated with L-carbocisteine and exposed to hydrogen peroxide (H2O2). Cell apoptosis was assessed using a cell death detection ELISA, and the pathways leading to cell apoptosis were examined by measurement of caspase-3 and caspase-9 by western blot analysis with fluorescent detection.
Results: The proportion of apoptotic cells in human tracheal epithelium was increased in a concentration- and time-dependent manner, following exposure to H2O2. Treatment with L-carbocisteine reduced the proportion of apoptotic cells. In contrast, H2O2 did not increase the concentration of LDH in supernatants of epithelial cells. Exposure to H2O2 activated caspase-3 and caspase-9, and L-carbocisteine inhibited the H2O2-induced activation of these caspases. L-carbocisteine activated Akt phosphorylation, which modulates caspase activation, and the inhibitors of Akt, LY294002 and wortmannin, significantly reversed the inhibitory effects of L-carbocisteine on H2O2-induced cell apoptosis.
Conclusions: These findings suggest that in human airway epithelium, L-carbocisteine may inhibit cell damage induced by H2O2 through the activation of Akt phosphorylation. L-carbocisteine may have antioxidant effects, as well as mucolytic activity, in inflamed airways.
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