{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T06:59:54Z","timestamp":1789628394051,"version":"build-2803163510"},"reference-count":48,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2018,7,2]],"date-time":"2018-07-02T00:00:00Z","timestamp":1530489600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004663","name":"Ministry of Science and Technology, Taiwan","doi-asserted-by":"publisher","award":["MOST 104\u20102320\u2010B\u2010040\u2010013"],"award-info":[{"award-number":["MOST 104\u20102320\u2010B\u2010040\u2010013"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Environmental Toxicology"],"published-print":{"date-parts":[[2018,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>IARC has classified the betel nut as a human environmental carcinogen. Previous studies have found that arecoline (AR) is the major alkaloid present in the saliva of betel quid chewers. Saliva contains a large content of AR which has been further shown to cause mutation of oral mucosa cells, resulting in oral cancer. Whereas, to date, there are only few studies reported the hepatotoxicity associated with arecoline and betel nut chewing. Therefore, the main purpose of this study was to determine the toxic effects of AR and its oxidative metabolite, arecoline <jats:italic>N<\/jats:italic>\u2010oxide (ARNO), in normal liver cell lines.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The cytotoxic, genotoxic, and mutagenic effects were detected by crystal violet staining, alkaline comet assay, and Salmonella mutagenicity test, respectively. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2\u2010DCFDA assay.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Our results demonstrated that ARNO exerted higher cytotoxicity, DNA damage, and mutagenicity than its parent compound arecoline in liver cells. Antioxidants, such as <jats:italic>N\u2010<\/jats:italic>acetylcysteine, Trolox, and penicillamine, strongly protected liver cells from ARNO\u2010induced DNA damage and ROS production. Furthermore, co\u2010treatment with Mito\u2010TEMPO also effectively blocked ARNO\u2010induced ROS production in liver cells. Besides antioxidants, co\u2010treatment with 1\u2010aminobenzotriazole and methimazole nearly completely suppressed ARNO\u2010induced ROS production in liver cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Our data suggest that arecoline ingested from the habit of chewing betel quid can be primarily oxidized to ARNO, thereby enhancing its toxicity through increased ROS production. Considering the excellent protective effects of both mitochondria\u2010targeted antioxidant and CYP450 inhibitor on ARNO\u2010induced ROS production in liver cells, mitochondria CYP450\u2010mediated metabolism of ARNO may be a key mechanism. Collectively, our results provide novel cellular evidence for the positive connection between habitual betel quid chewing and the risk for liver damage.<\/jats:p><\/jats:sec>","DOI":"10.1002\/tox.22588","type":"journal-article","created":{"date-parts":[[2018,7,2]],"date-time":"2018-07-02T10:48:49Z","timestamp":1530528529000},"page":"1029-1038","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":71,"title":["CYP450\u2010mediated mitochondrial ROS production involved in arecoline <i>N<\/i>\u2010oxide\u2010induced oxidative damage in liver cell lines"],"prefix":"10.1002","volume":"33","author":[{"given":"Tsu\u2010Shing","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Biomedical Sciences Chung Shan Medical University  Taichung Taiwan"},{"name":"Department of Medical Research Chung Shan Medical University Hospital  Taichung Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheng\u2010Ping","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences Chung Shan Medical University  Taichung Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu\u2010Pong","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences Chung Shan Medical University  Taichung Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mu\u2010Rong","family":"Chao","sequence":"additional","affiliation":[{"name":"Department of Occupational Safety and Health Chung Shan Medical University  Taichung Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chien\u2010Chun","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Nutrition Chung Shan Medical University  Taichung Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai\u2010Li","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Nutrition Chung Shan Medical University  Taichung Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,7,2]]},"reference":[{"key":"e_1_2_8_2_1","volume-title":"International Agency for Research on Cancer. 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