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.
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.
Bilbault
Pascal
P
Service de Réanimation Médicale, Hôpitaux Universitaires, and School of Medicine, University L. Pasteur, Strasbourg, France. pascal.bilbault@chru-strasbourg.fr
Lavaux
Thomas
T
Launoy
Anne
A
Gaub
Marie P
MP
Meyer
Nicolas
N
Oudet
Pierre
P
Pottecher
Thierry
T
Jaeger
Albert
A
Schneider
Francis
F
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
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