title: Amiodarone induces apoptosis of human and rat alveolar epithelial cells in vitro.
abstract: The antiarrhythmic amiodarone (AM) and its metabolite desethylamiodarone (Des) are known to cause AM-induced pulmonary toxicity, but the mechanisms underlying this disorder remain unclear. We hypothesized that AM might cause AM-induced pulmonary toxicity in part through the induction of apoptosis or necrosis in alveolar epithelial cells (AECs). Two models of type II pneumocytes, the human AEC-derived A549 cell line and primary AECs isolated from adult Wistar rats, were incubated with AM or Des for 20 h. Apoptotic cells were determined by morphological assessment of nuclear fragmentation with propidium iodide on ethanol-fixed cells. Necrotic cells were quantitated by loss of dye exclusion. Both AM and Des caused dose-dependent necrosis starting at 2.5 and 0.1 microg/ml, respectively, in primary rat AECs and at 10 and 5 microg/ml in subconfluent A549 cells (P < 0.05 and P < 0.01, respectively). AM and Des also induced dose-dependent apoptosis beginning at 2.5 microg/ml in the primary AECs (P < 0.05 for both compounds) and at 10 and 5 microg/ml, respectively, in the A549 cell line (P < 0.01). The two compounds also caused significant net cell loss (up to 80% over 20 h of incubation) by either cell type at drug concentrations near or below the therapeutic serum concentration for AM. The cell loss was not due to detachment but was blocked by the broad-spectrum caspase inhibitor Z-Val-Ala-Asp-fluoromethylketone. Furthermore, the angiotensin-converting enzyme inhibitor captopril (500 ng/ml) and the angiotensin-receptor antagonist saralasin (50 microg/ml) significantly inhibited both the induction of apoptosis and net cell loss in response to AM. These results are consistent with recent work from this laboratory demonstrating potent inhibition of apoptosis in human AECs by captopril (Uhal BD, Gidea C, Bargout R, Bifero A, Ibarra-Sunga O, Papp M, Flynn K, and Filippatos G. Am J Physiol Lung Cell Mol Physiol 275: L1013-L1017, 1998). They also suggested that the accumulation of AM and/or its primary metabolite Des in lung tissue may induce cytotoxicity of AECs that might be inhibitable by angiotensin-converting enzyme inhibitors or other antagonists of the renin-angiotensin system.
authors.affiliation: Department of Medicine, Cook County Hospital, Chicago, Illinois 60612, USA.
authors.forename: R
authors.initials: R
authors.lastname: Bargout
authors.forename: A
authors.initials: A
authors.lastname: Jankov
authors.forename: E
authors.initials: E
authors.lastname: Dincer
authors.forename: R
authors.initials: R
authors.lastname: Wang
authors.forename: T
authors.initials: T
authors.lastname: Komodromos
authors.forename: O
authors.initials: O
authors.lastname: Ibarra-Sunga
authors.forename: G
authors.initials: G
authors.lastname: Filippatos
authors.forename: B D
authors.initials: BD
authors.lastname: Uhal
mesh_terms: D000230:Adenocarcinoma; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000638:Amiodarone / Q000031:analogs & derivatives* / Q000494:pharmacology; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002216:Captopril / Q000494:pharmacology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D003603:Cytotoxins / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D008175:Lung Neoplasms; D008297:Male; D011650:Pulmonary Alveoli / Q000166:cytology*; D051381:Rats; D017208:Rats, Wistar; D012084:Renin-Angiotensin System / Q000502:physiology; D014407:Tumor Cells, Cultured
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D000806:Angiotensin-Converting Enzyme Inhibitors; D015853:Cysteine Proteinase Inhibitors; D003603:Cytotoxins; D004791:Enzyme Inhibitors; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D002216:Captopril; C036116:desethylamiodarone; D000638:Amiodarone