{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T16:21:13Z","timestamp":1780590073461,"version":"3.54.1"},"reference-count":36,"publisher":"Oxford University Press (OUP)","issue":"6","license":[{"start":{"date-parts":[[2020,11,25]],"date-time":"2020-11-25T00:00:00Z","timestamp":1606262400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"name":"Department of Pharmacy and Master Program"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,1,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Theophylline is a methylxanthine drug used in therapy for respiratory diseases. However, the impact of theophylline on Ca2+ signaling has not been explored in liver cells. This study examined whether theophylline affected Ca2+ homeostasis and its related cytotoxicity in AML12 mouse hepatocytes. Cell viability was measured by the cell viability reagent (WST-1). Cytosolic Ca2+ concentration ([Ca2+]i) was measured by the Ca2+-sensitive fluorescent dye fura-2. Theophylline (25\u2013125\u00a0\u03bcM) induced [Ca2+]i rises and cause cytotoxicity in AML12 cells. This cytotoxic response was reversed by chelation of cytosolic Ca2+ with BAPTA\/AM. In Ca2+-free medium, treatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin abolished theophylline-induced [Ca2+]i rises. Conversely, treatment with theophylline also abolished thapsigargin-induced [Ca2+]i rises. However, inhibition of PLC failed to alter theophylline-evoked [Ca2+]i rises. In Ca2+-containing medium, modulators of store-operated Ca2+ channels inhibited 30% of the [Ca2+]i rises, whereas the PKC modulators had no effect. Furthermore, theophylline-induced Ca2+ influx was confirmed by Mn2+-induced quench of fura-2 fluorescence. Together, in AML12 cells, theophylline caused Ca2+-associated cytotoxicity and induced Ca2+ entry through PLC-independent Ca2+ release from the endoplasmic reticulum and PKC-insensitive store-operated Ca2+ channels. BAPTA-AM with its protective effects may be a potential compound for prevention of theophylline-induced cytotoxicity.<\/jats:p>","DOI":"10.1093\/toxres\/tfaa084","type":"journal-article","created":{"date-parts":[[2020,10,19]],"date-time":"2020-10-19T19:15:04Z","timestamp":1603134904000},"page":"790-797","source":"Crossref","is-referenced-by-count":2,"title":["Mechanism of a methylxanthine drug theophylline-induced Ca2+ signaling and cytotoxicity in AML12 mouse hepatocytes"],"prefix":"10.1093","volume":"9","author":[{"given":"Gwo-Ching","family":"Sun","sequence":"first","affiliation":[{"name":"Department of Anesthesiology, Kaohsiung Medical University Hospital, Kaohsiung, 80708, Taiwan"},{"name":"Department of Anesthesiology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, 80756, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei-Zhe","family":"Liang","sequence":"additional","affiliation":[{"name":"Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan"},{"name":"Department of Pharmacy and Master Program, College of Pharmacy and Health Care, Tajen University, No.20, Weixin Rd., Yanpu Township, Pingtung County 90741, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2020,11,25]]},"reference":[{"key":"2021010603301629200_ref1","doi-asserted-by":"crossref","first-page":"901","DOI":"10.1164\/rccm.201302-0388PP","article-title":"Theophylline","volume":"188","author":"Barnes","year":"2013","journal-title":"Am J Respir Crit Care Med"},{"key":"2021010603301629200_ref2","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0050019","article-title":"Spectral analysis of naturally occurring methylxanthines (theophylline, theobromine and caffeine) binding with DNA","volume":"7","author":"Johnson","year":"2012","journal-title":"PLoS One"},{"key":"2021010603301629200_ref3","doi-asserted-by":"crossref","first-page":"1504","DOI":"10.1183\/09031936.00131513","article-title":"Impact of theophylline\/corticosteroid combination therapy on sputum hydrogen sulfide levels in patients with COPD","volume":"43","author":"Kirkham","year":"2014","journal-title":"Eur Respir J"},{"key":"2021010603301629200_ref4","doi-asserted-by":"crossref","first-page":"231","DOI":"10.5415\/apallergy.2013.3.4.231","article-title":"CYP1A2 polymorphism and theophylline clearance in Korean non-smoking asthmatics","volume":"3","author":"Yim","year":"2013","journal-title":"Asia Pac Allergy"},{"key":"2021010603301629200_ref5","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1023\/A:1026508605951","article-title":"Antimitotic and cytotoxic effects of theophylline in MDA-MB-231 human breast cancer cells","volume":"64","author":"Slotkin","year":"2000","journal-title":"Breast Cancer Res Treat"},{"key":"2021010603301629200_ref6","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1093\/oxfordjournals.jbchem.a003197","article-title":"Theophylline-induced apoptosis is paralleled by protein kinase A-dependent tissue transglutaminase activation in cancer cells","volume":"132","author":"Caraglia","year":"2002","journal-title":"J Biochem"},{"key":"2021010603301629200_ref7","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1186\/1743-7075-9-76","article-title":"Theophylline is able to partially revert cachexia in tumour-bearing rats","volume":"9","author":"Olivan","year":"2012","journal-title":"Nutr Metab (Lond)"},{"key":"2021010603301629200_ref8","doi-asserted-by":"crossref","first-page":"1261","DOI":"10.1152\/physrev.00006.2016","article-title":"The inositol trisphosphate\/calcium signaling pathway in health and disease","volume":"96","author":"Berridge","year":"2016","journal-title":"Physiol Rev"},{"key":"2021010603301629200_ref9","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1113\/jphysiol.2013.257527","article-title":"Calcium regulation of neural rhythms, memory and Alzheimer's disease","volume":"592","author":"Berridge","year":"2014","journal-title":"J Physiol"},{"key":"2021010603301629200_ref10","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1042\/bst0310930","article-title":"Cardiac calcium signalling","volume":"31","author":"Berridge","year":"2003","journal-title":"Biochem Soc Trans"},{"key":"2021010603301629200_ref11","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/S0960-9822(94)00041-2","article-title":"Intracellular calcium. 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