title: Impairment of autophagosome-lysosome fusion contributes to chronic ethanol-induced liver injury.
abstract: The pathogenic mechanism underlying alcoholic fatty liver (AFL) is not clear. Autophagy is a self-digestion process that is critical for the maintenance of cellular homeostasis and regulation of lipid metabolism. We investigated the role of autophagy and autophagic flux in hepatic injury induced by chronic ethanol feeding in mice. C57BL/6 mice were fed a Lieber-DeCarli ethanol diet (ED) to induce AFL or an isocaloric control diet for 6 weeks. Chloroquine (CQ, 10 mg/kg, intra-peritoneally [i.p.]) or rapamycin (Rapa, 5 mg/kg, i.p.) were administered during the last 2 weeks of the experimental period. Chronic ethanol feeding induced AFL with focal necrosis associated with increased levels of hepatic triglyceride. This phenomenon was aggravated by CQ, an inhibitor of autophagy, and attenuated by Rapa, an inducer of autophagy. Expression of microtubule-associated protein 1 light chain 3 (LC3)-II and sequestosome1/p62 significantly increased in the ED group. Moreover, accumulation of autophagosomes was observed by transmission electron microscopy in chronic ethanol-treated mice. Chronic ethanol consumption decreased protein expression of LC3 lipidation-related proteins Atg3 and Atg7, and the lysosomal proteins lysosome-associated membrane protein-2 and Rab7, and increased the protein expression of calpain 1 and phosphorylated mammalian target of rapamycin. Taken together, these findings suggest that chronic ethanol consumption leads to impairment of autophagic flux, which contributes to ethanol-induced liver injury.
authors.affiliation: School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Hong-Ik
authors.initials: HI
authors.lastname: Cho
authors.affiliation: School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Joo-Wan
authors.initials: JW
authors.lastname: Choi
authors.affiliation: School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea. Electronic address: sunmee@skku.edu.
authors.forename: Sun-Mee
authors.initials: SM
authors.lastname: Lee
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001343:Autophagy / Q000187:drug effects* / Q000502:physiology; D002738:Chloroquine / Q000494:pharmacology; D000431:Ethanol / Q000009:adverse effects; D008099:Liver / Q000187:drug effects / Q000648:ultrastructure; D008108:Liver Diseases, Alcoholic / Q000209:etiology*; D008247:Lysosomes / Q000187:drug effects* / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission; D020123:Sirolimus / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D000431:Ethanol; D002738:Chloroquine; D020123:Sirolimus
keywords: Alcoholic fatty liver; Autolysosome; Autophagic flux; Autophagosome; Chronic ethanol consumption; LC3 lipidation