Autophagy: a cyto-protective mechanism which prevents primary human hepatocyte apoptosis during oxidative stress.
The role of autophagy in the response of human hepatocytes to oxidative stress remains unknown. Understanding this process may have important implications for the understanding of basic liver epithelial cell biology and the responses of hepatocytes during liver disease. To address this we isolated primary hepatocytes from human liver tissue and exposed them ex vivo to hypoxia and hypoxia-reoxygenation (H-R). We showed that oxidative stress increased hepatocyte autophagy in a reactive oxygen species (ROS) and class III PtdIns3K-dependent manner. Specifically, mitochondrial ROS and NADPH oxidase were found to be key regulators of autophagy. Autophagy involved the upregulation of BECN1, LC3A, Atg7, Atg5 and Atg 12 during hypoxia and H-R. Autophagy was seen to occur within the mitochondria of the hepatocyte and inhibition of autophagy resulted in the lowering a mitochondrial membrane potential and onset of cell death. Autophagic responses were primarily observed in the large peri-venular (PV) hepatocyte subpopulation. Inhibition of autophagy, using 3-methyladenine, increased apoptosis during H-R. Specifically, PV human hepatocytes were more susceptible to apoptosis after inhibition of autophagy. These findings show for the first time that during oxidative stress autophagy serves as a cell survival mechanism for primary human hepatocytes.
Centre for Liver Research, The Institute for Biomedical Research, The Medical School, University of Birmingham, Birmingham, West Midlands UK. balsin@hotmail.com
Ricky H
RH
Bhogal
Christopher J
CJ
Weston
Stuart M
SM
Curbishley
David H
DH
Adams
Simon C
SC
Afford
D017209:Apoptosis; D001343:Autophagy; D015687:Cell Hypoxia; D002469:Cell Separation; D048429:Cell Size; D002478:Cells, Cultured; D019610:Cytoprotection; D022781:Hepatocytes / Q000201:enzymology / Q000473:pathology*; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D008107:Liver Diseases / Q000473:pathology; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D008954:Models, Biological; D019255:NADPH Oxidases / Q000378:metabolism; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
D008869:Microtubule-Associated Proteins; D017382:Reactive Oxygen Species; D019255:NADPH Oxidases; D010100:Oxygen