title: Hypoxia and reoxygenation of primary human hepatocytes induce proteome changes of glucose metabolism, oxidative protection and peroxisomal function.
abstract: Protective hepatocellular responses to a hypoxic challenge are crucial to preserve liver function. The knowledge of affected metabolic functions could help assess and enhance hepatic ischemic tolerance. Here we studied adaptive mechanisms in human hepatocytes after hypoxia and reoxygenation using a proteomic approach. Proteins from primary hepatocytes were extracted after 6 h of hypoxia and 24 h of reoxygenation. The proteome was analyzed by 2D-electrophoresis. Densitometry and mass spectrometry (MALDI-TOF-MS) were used for protein identification. Two hundred and sixty-two spots were differentially analyzed and 33 spots displayed significant differences between hypoxic and normoxic cells. Seventeen proteins were identified by mass spectrometry. After hypoxia and reoxygenation the UTP-glucose-1-phosphate uridyltransferase, phosphoglycerate kinase1, fructose-1,6-bisphosphate aldolase, glyceraldehyde-3-phosphate dehydrogenase, fructose-1,6-bisphosphatase, thiosulfat-sulfurtransferase, thioredoxin peroxidase, peroxiredoxin III, and annexin A2 proteins were down-regulated. An increased expression was found for carbamoyl phosphate synthetase I, heat shock 70 kDa protein5, phosphoenolpyruvate carboxy-kinase, catalase isoform2, peroxiredoxin II, glutathione S-transferase, hydroxyacid oxidase1, and F1-ATP synthase, alpha subunit1. Hepatocellular adaptation to hypoxia and reoxygenation involve glucose metabolism, peroxisomal functions, and oxidative stress protection. The identified proteins can serve as possible diagnostic targets to monitor hepatic hypoxic tolerance e.g. in the context of liver surgery and transplantation.
authors.affiliation: Department of General and Vascular Surgery, Johann Wolfgang Goethe-University Frankfurt, Medical School, Frankfurt, Germany. strey@gmx.de
authors.forename: Christoph W
authors.initials: CW
authors.lastname: Strey
authors.forename: Johannes
authors.initials: J
authors.lastname: Gestrich
authors.forename: Tobias
authors.initials: T
authors.lastname: Beckhaus
authors.forename: Rosa Maria
authors.initials: RM
authors.lastname: Marquez-Pinilla
authors.forename: Elsie
authors.initials: E
authors.lastname: Oppermann
authors.forename: Christian
authors.initials: C
authors.lastname: Mönch
authors.forename: John D
authors.initials: JD
authors.lastname: Lambris
authors.forename: Michael
authors.initials: M
authors.lastname: Karas
authors.forename: Wolf O
authors.initials: WO
authors.lastname: Bechstein
mesh_terms: D000368:Aged; D015687:Cell Hypoxia; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005260:Female; D005947:Glucose / Q000378:metabolism*; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D045022:Lactate Dehydrogenases / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D020675:Peroxisomes / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D020543:Proteome; D045022:Lactate Dehydrogenases; D005947:Glucose; D010100:Oxygen