Inflammatory and apoptotic alterations in serum and injured tissue after experimental polytrauma in mice: distinct early response compared with single trauma or "double-hit" injury.
BACKGROUND
The exact alterations of the immune system after polytrauma leading to sepsis and multiple-organ failure are poorly understood. Thus, the early local and systemic inflammatory and apoptotic response was characterized in a new polytrauma model and compared with the alterations seen after single or combined injuries.


METHODS
Anesthetized C57BL/6 mice were subjected to either blunt bilateral chest trauma (Tx), closed head injury, right femur fracture including contralateral soft tissue injury, or a combination of injuries (PTx). After 2 hours or 6 hours, animals were sacrificed, and the systemic as well as the local pulmonary immune response (bronchoalveolar lavage [BAL]/plasma cytokines, lung myeloperoxidase [MPO] activity, and alveolocapillary barrier dysfunction) were evaluated along with lung/brain apoptosis (lung caspase 3 Western blotting, immunohistochemistry, and polymorphonuclear leukocytes [PMN] Annexin V).


RESULTS
Hemoglobin, PO2 saturation, and pH did not differ between the experimental groups. Local BAL cytokines/chemokines were significantly increased in almost all groups, which included Tx. There was no further enhancement of this local inflammatory response in the lungs in case of PTx. At 2 hours, all groups except sham and closed head injury alone revealed an increased activity of lung MPO. However, 6 hours after injury, lung MPO remained increased only in the PTx group. Increased BAL protein levels were found, reflecting enhanced lung leakage in all groups with Tx 6 hours after trauma. Only after PTx was neutrophil apoptosis significantly decreased, whereas lung caspase 3 and plasma interleukin 6/keratinocyte chemoattractant (KC) were substantially increased.


CONCLUSIONS
The combination of different injuries leads to an earlier systemic inflammatory response when compared with the single insults. Interestingly, only after PTx but not after single or double hits was lung apoptosis increased, and PMN apoptosis was decreased along with a prolonged presence of neutrophils in the lungs, which may therefore represent a possible pathomechanism for lung injury after polytrauma.
Department of Trauma-, Hand-, Plastic- and Reconstructive Surgery, University of Ulm, 89075 Ulm, Germany.
Sebastian
S
Weckbach
Christoph
C
Hohmann
Sonja
S
Braumueller
Stephanie
S
Denk
Bettina
B
Klohs
Philip F
PF
Stahel
Florian
F
Gebhard
Markus S
MS
Huber-Lang
Mario
M
Perl
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D015153:Blotting, Western; D001921:Brain / Q000276:immunology / Q000503:physiopathology; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D053148:Caspase 3 / Q000378:metabolism; D018925:Chemokines / Q000032:analysis / Q000097:blood; D016207:Cytokines / Q000032:analysis / Q000097:blood; D005434:Flow Cytometry; D006454:Hemoglobins / Q000032:analysis; D007249:Inflammation / Q000097:blood / Q000209:etiology* / Q000276:immunology / Q000503:physiopathology; D008168:Lung / Q000737:chemistry / Q000276:immunology / Q000503:physiopathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009104:Multiple Trauma / Q000097:blood / Q000150:complications* / Q000276:immunology / Q000503:physiopathology; D009195:Peroxidase / Q000378:metabolism; D014947:Wounds and Injuries / Q000097:blood / Q000150:complications* / Q000276:immunology / Q000503:physiopathology
D018925:Chemokines; D016207:Cytokines; D006454:Hemoglobins; D009195:Peroxidase; D053148:Caspase 3