title: Pivotal role of glutathione depletion in plasma-induced endothelial oxidative stress during sepsis.
abstract: OBJECTIVE
  Plasma from septic shock patients can induce production of reactive oxygen species (ROS) by human umbilical vein endothelial cells (HUVEC) in vitro. How endothelial cells defend themselves against ROS under increased oxidative stress has not yet been examined. This study investigates the antioxidant defenses of HUVEC exposed to plasma obtained from either septic shock patients or healthy volunteers.
  
  
  METHODS
  Prospective, observational study.
  
  
  METHODS
  Medical intensive care unit in a university hospital.
  
  
  METHODS
  Twenty-five patients with septic shock and 10 healthy volunteers.
  
  
  METHODS
  Blood samples were collected within the first 24 hrs of septic shock. In vitro HUVEC production of ROS was studied by spectrofluorimetry using 2',7'-dichlorodihydrofluorescein diacetate fluorescent dye. Reactive nitrogen species were also assessed. Intracellular reduced glutathione (GSH) levels were measured using monochlorobimane fluorescent dye. Activity of catalase and superoxide dismutase in HUVEC were also measured. Cell death was assessed using YOPRO fluorescent dye and the MTT assay.
  
  
  RESULTS
  On admission, the septic shock population's mean age was 55 yrs old, the mean Sequential Organ Failure Assessment score was 12, mean simplified acute physiology score was 50, and intensive care unit mortality rate was 45%. Evaluation of HUVEC antioxidant defenses showed a significantly decreased GSH level, increased catalase activity, and unchanged superoxide dismutase activity. ROS levels and cell death were significantly reduced when cells were pretreated with N-acetylcysteine or GSH, but no changes in reactive nitrogen species were observed.
  
  
  CONCLUSIONS
  This study demonstrates that plasma-induced ROS production by HUVEC is associated with an intracellular decrease in reduced GSH. Both ROS levels and cell death decreased when N-acetylcysteine or GSH were added before exposing the cells to plasma. These data suggest a pivotal role of alterations in GSH in damage caused by sepsis-generated ROS in endothelial cell.
authors.affiliation: Département d'anesthésie réanimation, Hôpital Bicêtre, Assistance Publique des Hopitaux de Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Huet
authors.forename: Christaine
authors.initials: C
authors.lastname: Cherreau
authors.forename: Carole
authors.initials: C
authors.lastname: Nicco
authors.forename: Laurent
authors.initials: L
authors.lastname: Dupic
authors.forename: Marc
authors.initials: M
authors.lastname: Conti
authors.forename: Didier
authors.initials: D
authors.lastname: Borderie
authors.forename: Frédéric
authors.initials: F
authors.lastname: Pene
authors.forename: Eric
authors.initials: E
authors.lastname: Vicaut
authors.forename: Dan
authors.initials: D
authors.lastname: Benhamou
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Mira
authors.forename: Jacques
authors.initials: J
authors.lastname: Duranteau
authors.forename: Frédéric
authors.initials: F
authors.lastname: Batteux
mesh_terms: D018806:APACHE; D000111:Acetylcysteine / Q000494:pharmacology; D000328:Adult; D000368:Aged; D016923:Cell Death / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D016166:Free Radical Scavengers / Q000494:pharmacology; D005978:Glutathione / Q000172:deficiency* / Q000502:physiology*; D006801:Humans; D007362:Intensive Care Units; D008297:Male; D008875:Middle Aged; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012772:Shock, Septic / Q000097:blood* / Q000378:metabolism / Q000401:mortality
chemical_list: D016166:Free Radical Scavengers; D017382:Reactive Oxygen Species; D005978:Glutathione; D000111:Acetylcysteine