Dynamics of neutrophilic NF-kB translocation in relation to IL-8 mRNA expression after major trauma.
OBJECTIVE
Systemic inflammation subsequent to polytrauma is connected to neutrophil (PMN) dysregulation characterized by reduced NF-kB-translocation and cytokine expression. The dynamics of NF-kB-activation as well as its down-stream regulation of IL-8-expression in PMN following major trauma remain unclear. The aim of this pilot study was to analyse NF-kB nuclear translocation in relation to IL- 8-mRNA-expression in PMN after major trauma.


METHODS
PMN were isolated from blood samples of 15 major trauma patients (New Injury Severity Score, NISS > 16) drawn within 90 min and subsequently 6, 12, 24, 48, 72 h after trauma. NF-kB-translocation was analysed by Electrophoretic Mobility Shift Assay, EMSA and quantified by densitometry [arbitrary units], IL-8-mRNA-expression by RT-PCR, [copies/50 ng RNA]. Additionally, NF-kB-translocation and IL-8-expression in PMN of healthy volunteers were analysed natively (-control) and after LPS stimulation (+control).


RESULTS
NF-kB-translocation and IL-8-mRNA-expression was significantly increased in polytrauma patients (n=15; NISS: 34 +/- 8 [mean +/- SEM]) initially. In non-survivors, NFkB- translocation was significantly increased on admission and subsequently reduced within 6 h, while it increased in the survivors group. After 24 h, a second significant increase in NF-kB-activity and IL-8-expression was found in survivors that was subsequently reduced in both groups.


CONCLUSIONS
This pilot study has shown that a concomitant initial increase in transcriptional NF-kB-activity and IL-8 mRNA expression was observed in the early posttraumatic period which preceded the down-regulation of the innate immune system.
Stegmaier
J C
JC
Department of Traumatology and Orthopedic Surgery, Ludwig-Maximilians-University Munich, Munich, Germany. julia.stegmaier@med.uni-muenchen.de
Kirchhoff
C
C
Bogner
V
V
Matz
M
M
Kanz
K-G
KG
Mutschler
W
W
Biberthaler
P
P
D000293:Adolescent; D000328:Adult; D000368:Aged; D005260:Female; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009102:Multiple Organ Failure / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D009504:Neutrophils / Q000378:metabolism*; D021381:Protein Transport; D012333:RNA, Messenger / Q000032:analysis*; D014947:Wounds and Injuries / Q000378:metabolism*
D016209:Interleukin-8; D016328:NF-kappa B; D012333:RNA, Messenger