beta2 Adrenoreceptor blockade attenuates the hyperinflammatory response induced by traumatic injury.
BACKGROUND
Major operative injury initiates immunologic changes that may result in a systemic inflammatory response, leading to organ dysfunction/sepsis. Catecholamines seem to be key mediators. The effects of beta(2) receptor blockade in vitro and in vivo before and after operative injury were studied to clarify their role.


METHODS
In vitro studies were done in RAW 264.7 cells using epinephrine (50 micromol/L) with or without alpha(2)- and beta(2)-receptor blockade. Comparative gene expression analysis on the Toll-like receptor (TLR)-4 receptor signaling pathway was performed between RAW cells pretreated with epinephrine with subsequent lipopolysaccharide (LPS) stimulation versus LPS alone. Confirmatory studies were performed by real-time reverse transcription polymerase chain reaction (RT-PCR). Tumor necrosis factor (TNF)-alpha gene and protein expression were determined via real time RT-PCR and enzyme-linked immunosorbent assay, respectively. In vivo, Balb/C mice received no treatment nor injury (group I). Group II received operative injury and vehicle injection, group III received ICI 118,551 (beta(2)-receptor antagonist) 30 minutes before or in separate studies after operative injury. At 7 days, splenic macrophages were harvested and cytokine production was measured with or without LPS. In separate experiments, cecal ligation and puncture (CLP) was done 7 days after operation and survival determined.


RESULTS
In vitro studies demonstrated that epinephrine pretreatment significantly increased TNF-alpha production with LPS stimulation (P < .05). beta(2)-Receptor blockade significantly attenuated (P < .05) LPS stimulated TNF-alpha production. MD-2, an essential coactivator of TLR-4 signaling, gene expression was significantly elevated when cells were pretreated with epinephrine before LPS exposure (P < .001). In vivo studies demonstrated a significant decrease (P < .05) in TNF-alpha and interleukin-6 production in the ICI 118,551 group. Similar findings were demonstrated measuring monocyte chemoattractant protein-1 and interferon-gamma cytokine levels (P < .05) versus no treatment. ICI 118,551 treatment 30 minutes before operation demonstrated a 31% reduction in mortality after CLP (P < .05).


CONCLUSIONS
This study demonstrates that beta(2)-receptor blockade reduces macrophage cytokine production and improves survival showing the critical importance of catecholamines to the immunologic response in surgery.
Rough
James
J
Department of Surgery, Temple University School of Medicine, Philadelphia, PA 19140, USA.
Engdahl
Ryan
R
Opperman
Kay
K
Yerrum
Smitha
S
Monroy
M Alexandra
MA
Daly
John M
JM
D058672:Adrenergic beta-2 Receptor Antagonists; D000818:Animals; D045744:Cell Line, Tumor; D004837:Epinephrine / Q000008:administration & dosage / Q000378:metabolism*; D005260:Female; D006470:Hemorrhage / Q000150:complications; D008070:Lipopolysaccharides; D051218:Lymphocyte Antigen 96 / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D009104:Multiple Trauma / Q000150:complications / Q000188:drug therapy / Q000276:immunology*; D011412:Propanolamines / Q000627:therapeutic use*; D018746:Systemic Inflammatory Response Syndrome / Q000209:etiology / Q000276:immunology* / Q000517:prevention & control; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D015854:Up-Regulation
D058672:Adrenergic beta-2 Receptor Antagonists; D008070:Lipopolysaccharides; C411728:Ly96 protein, mouse; D051218:Lymphocyte Antigen 96; D011412:Propanolamines; D014409:Tumor Necrosis Factor-alpha; C026777:ICI 118551; D004837:Epinephrine