Dynamics of neutrophilic NF-kB translocation in relation to IL-8 mRNA expression after major trauma

Birkhuser Verlag, Basel, 2008

Inamm. res. 57 (2008) 5475541023-3830/08/110547-8DOI 10.1007/s00011-008-7207-6 Inammation Research

Dynamics of neutrophilic NF-kB translocation in relation to IL-8 mRNA expression after major trauma

J. C. Stegmaier1, C. Kirchhoff2, V. Bogner1, M. Matz1, K.-G. Kanz1, W. Mutschler1 and P. Biberthaler1

1 Department of Traumatology and Orthopedic Surgery, Ludwig-Maximilians-University Munich, Germany, Fax: [phone],

e-mail: [email protected]

2 Department of Orthopaedic Sports Surgery, Klinikum rechts der Isar, Technische Universitt, Ismaningerstrasse 22, 81675 Munich, Germany

Received 27 November 2007; returned for revision 10 May 2008; received from nal revision 15 July 2008; accepted by M. Parnham 16 July 2008

Published Online First 13 November 2008

Abstract. Objective: Systemic inammation 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.

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

Key words: NF-kB EMSA RT-PCR IL-8 PMN NISS major trauma MOF

Introduction

Mortality after major trauma is associated with insufciency of the innate immune system [36]. Early destabilisation of the innate immune system after trauma has been described as Systemic Inammatory Response Syndrome, SIRS...