title: Influence of drotrecogin alpha (activated) infusion on the variation of Bax/Bcl-2 and Bax/Bcl-xl ratios in circulating mononuclear cells: a cohort study in septic shock patients.
abstract: OBJECTIVE
  Drotrecogin alpha (activated) (DAA), or recombinant human activated protein C, is a new treatment in sepsis-induced multiple organ failure, leading to significant reduction in the mortality rate, thanks to its anticoagulant properties. It has been suggested that DAA has anti-inflammatory and antiapoptotic effects in sepsis animal models. This study investigates the potential actions of DAA on circulating mononuclear cells apoptosis in human septic shock.
  
  
  METHODS
  Prospective, cohort study.
  
  
  METHODS
  Two intensive care wards and two research laboratories in a university hospital.
  
  
  METHODS
  Twenty-two septic shock patients with DAA treatment (DAA+), 19 septic shock patients without DAA treatment (DAA-), and 14 healthy controls were successively enrolled, but only 20 DAA+ and 16 DAA- patients fulfilled criteria for statistical analysis.
  
  
  METHODS
  Blood samples were collected at inclusion and 24 hrs later.
  
  
  RESULTS
  Circulating mononuclear cell apoptosis levels were assessed by flow cytometry with annexin V, and variations of the apoptotic rheostats (Bax/Bcl-2 and Bax/Bcl-xl ratios) were analyzed by real-time reverse transcription-polymerase chain reaction. Apoptosis was significantly increased in septic shock patients (DAA+, 12 +/- 6.4%; DAA-, 10.4 +/- 5%) vs. healthy patients (3.4 +/- 2.1%, p < .001). Twenty-four hours after DAA infusion, apoptosis was significantly lower in the DAA+ group compared with DAA- ones (respectively, 11.7 +/- 5.3% and 16.2 +/- 7.6%, p < .001). At inclusion, DAA+ and DAA- groups showed comparable Bax/Bcl-2 ratio (DAA+, 0.92 +/- 0.9; DAA-, 1.32 +/- 0.87) and Bax/Bcl-xl ratio (DAA+, 2 +/- 1.04; DAA-, 1.31 +/- 0.93). In contrast, 24 hrs later we observed a significant decrease in these ratios, indicating an antiapoptotic effect in the DAA+ group (Bax/Bcl-2, 0.39 +/- 0.27; Bax/Bcl-xl, 0.68 +/- 0.35) compared with the DAA- group (Bax/Bcl-2, 1.81 +/- 1.1; Bax/Bcl-xl, 1.22 +/- 0.92, p = .001 and p = .039, respectively).
  
  
  CONCLUSIONS
  In vivo, in human septic shock, DAA has antiapoptotic effects on circulating mononuclear cells, assessed by a significant decrease of both the Bax/Bcl-2 and Bax/Bcl-xl ratios.
authors.affiliation: Service de Réanimation Médicale, Hôpitaux Universitaires, and School of Medicine, University L. Pasteur, Strasbourg, France. pascal.bilbault@chru-strasbourg.fr
authors.forename: Pascal
authors.initials: P
authors.lastname: Bilbault
authors.forename: Thomas
authors.initials: T
authors.lastname: Lavaux
authors.forename: Anne
authors.initials: A
authors.lastname: Launoy
authors.forename: Marie P
authors.initials: MP
authors.lastname: Gaub
authors.forename: Nicolas
authors.initials: N
authors.lastname: Meyer
authors.forename: Pierre
authors.initials: P
authors.lastname: Oudet
authors.forename: Thierry
authors.initials: T
authors.lastname: Pottecher
authors.forename: Albert
authors.initials: A
authors.lastname: Jaeger
authors.forename: Francis
authors.initials: F
authors.lastname: Schneider
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000704:Analysis of Variance; D000890:Anti-Infective Agents / Q000494:pharmacology / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007262:Infusions, Intravenous; D007958:Leukocyte Count; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D008126:Lod Score; D008297:Male; D008875:Middle Aged; D011446:Prospective Studies; D011486:Protein C / Q000494:pharmacology / Q000627:therapeutic use*; D011994:Recombinant Proteins / Q000494:pharmacology / Q000627:therapeutic use; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012772:Shock, Septic / Q000097:blood / Q000188:drug therapy; D051028:bcl-2-Associated X Protein / Q000032:analysis / Q000187:drug effects* / Q000235:genetics; D051020:bcl-X Protein / Q000032:analysis / Q000187:drug effects* / Q000235:genetics
chemical_list: D000890:Anti-Infective Agents; C494811:BCL2L1 protein, human; D011486:Protein C; D011994:Recombinant Proteins; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; C421124:drotrecogin alfa activated