CYP3A-mediated apoptosis of dauricine in cultured human bronchial epithelial cells and in lungs of CD-1 mice

• Published in: Toxicology and applied pharmacology 2012-06, Vol.261 (3), p.248-254
• Main Authors: Jin, Hua, Shen, Shuijie, Chen, Xiaoyan, Zhong, Dafang, Zheng, Jiang
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; Animals; APOPTOSIS; Apoptosis - drug effects; bcl-Associated Death Protein - metabolism; BENZOQUINONES; Benzylisoquinolines - toxicity; Biological and medical sciences; Blotting, Western; Bronchi - cytology; Bronchi - drug effects; Buthionine Sulfoximine - pharmacology; Caspase 3 - metabolism; CELL CULTURES; Cell Survival - drug effects; Cells, Cultured; CYP3A; Cytochrome P-450 CYP3A - physiology; Cytochrome P-450 CYP3A Inhibitors; Cytochromes c - metabolism; Cytotoxicity; Dauricine; Enzyme Inhibitors - pharmacology; Epithelial cells; Epithelial Cells - drug effects; Flow Cytometry; GLUTATHIONE; Glutathione - metabolism; GSH depletion; Humans; In Situ Nick-End Labeling; INJURIES; Ketoconazole; Ketoconazole - pharmacology; L-Buthionine sulfoximine; Lung; Lung - cytology; Lung - drug effects; LUNGS; Male; Medical sciences; Menispermum; METABOLIC ACTIVATION; MICE; Quinone; Roots; Tetrahydroisoquinolines - toxicity; TOXICITY; Toxicology
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1016/j.taap.2012.03.025

Dauricine is the major bioactive component isolated from the root of Menispermum dauricum DC and has shown promising pharmacologic activities with a great potential for clinical use. Recently, we found that intraperitoneal exposure of dauricine produced selective pulmonary injury in mice. A quinone methide metabolite of dauricine was identified and is suggested to be associated with the pulmonary toxicity of dauricine. The present study evaluated the apoptotic effect of dauricine in cultured cells and mice, determined the change in cellular glutathione (GSH) contents after exposure to dauricine, investigated the role of GSH depletion in dauricine-induced cytotoxicity and apoptosis, and examined the role of CYP3A in dauricine-induced GSH depletion and apoptosis. Dauricine was found to induce apoptosis in NL-20 cells. Additionally, intraperitoneal administration of dauricine caused GSH depletion and apoptosis in lungs of mice. Treatment with ketoconazole, an inhibitor of CYP3A, reversed cellular GSH depletion in lungs of mice given dauricine and showed protective effect on dauricine-induced apoptosis in lungs of mice. This indicates that metabolic activation is involved in dauricine-induced GSH-depletion, cytotoxicity and apoptosis. The glutathione depletor l-buthionine sulfoximine showed potentiating effect on cytotoxicity and apoptosis induced by dauricine. We propose that dauricine is metabolized to a quinone methide intermediate which depletes cellular GSH, and the depletion of GSH may trigger and/or intensify the cytotoxicity and apoptosis induced by dauricine. ► Dauricine induced apoptosis in lungs in mice and in cultured human pulmonary cells. ► Dauricine depleted cellular GSH in lungs of mice and in the human pulmonary cells. ► CYP3A subfamily mediated GSH depletion and apoptosis induced by dauricine. ► l-Buthionine sulfoximine potentiated dauricine-induced GSH depletion and apoptosis.