title: Ropivacaine attenuates endotoxin plus hyperinflation-mediated acute lung injury via inhibition of early-onset Src-dependent signaling.
abstract: BACKGROUND
  Acute lung injury (ALI) is associated with high mortality due to the lack of effective therapeutic strategies. Mechanical ventilation itself can cause ventilator-induced lung injury. Pulmonary vascular barrier function, regulated in part by Src kinase-dependent phosphorylation of caveolin-1 and intercellular adhesion molecule-1 (ICAM-1), plays a crucial role in the development of protein-/neutrophil-rich pulmonary edema, the hallmark of ALI. Amide-linked local anesthetics, such as ropivacaine, have anti-inflammatory properties in experimental ALI. We hypothesized ropivacaine may attenuate inflammation in a "double-hit" model of ALI triggered by bacterial endotoxin plus hyperinflation via inhibition of Src-dependent signaling.
  
  
  METHODS
  C57BL/6 (WT) and ICAM-1 (-/-) mice were exposed to either nebulized normal saline (NS) or lipopolysaccharide (LPS, 10 mg) for 1 hour. An intravenous bolus of 0.33 mg/kg ropivacaine or vehicle was followed by mechanical ventilation with normal (7 ml/kg, NTV) or high tidal volume (28 ml/kg, HTV) for 2 hours. Measures of ALI (excess lung water (ELW), extravascular plasma equivalents, permeability index, myeloperoxidase activity) were assessed and lungs were homogenized for Western blot analysis of phosphorylated and total Src, ICAM-1 and caveolin-1. Additional experiments evaluated effects of ropivacaine on LPS-induced phosphorylation/expression of Src, ICAM-1 and caveolin-1 in human lung microvascular endothelial cells (HLMVEC).
  
  
  RESULTS
  WT mice treated with LPS alone showed a 49% increase in ELW compared to control animals (p = 0.001), which was attenuated by ropivacaine (p = 0.001). HTV ventilation alone increased measures of ALI even more than LPS, an effect which was not altered by ropivacaine. LPS plus hyperinflation ("double-hit") increased all ALI parameters (ELW, EVPE, permeability index, MPO activity) by 3-4 fold compared to control, which were again decreased by ropivacaine. Western blot analyses of lung homogenates as well as HLMVEC treated in culture with LPS alone showed a reduction in Src activation/expression, as well as ICAM-1 expression and caveolin-1 phosphorylation. In ICAM-1 (-/-) mice, neither addition of LPS to HTV ventilation alone nor ropivacaine had an effect on the development of ALI.
  
  
  CONCLUSIONS
  Ropivacaine may be a promising therapeutic agent for treating the cause of pulmonary edema by blocking inflammatory Src signaling, ICAM-1 expression, leukocyte infiltration, and vascular hyperpermeability.
authors.affiliation: Department of Anesthesiology, University of Illinois Hospital > Health Sciences System, 835 S. Wolcott Ave (m/c 868), Chicago, IL 60612, USA ; Institute of Anesthesiology, University Hospital Zurich, Zurich, Switzerland.
authors.forename: Tobias
authors.initials: T
authors.lastname: Piegeler
authors.affiliation: Department of Anesthesiology, University of Illinois Hospital > Health Sciences System, 835 S. Wolcott Ave (m/c 868), Chicago, IL 60612, USA ; Department of Pharmacology, University of Illinois Hospital > Health Sciences System, Chicago, IL, USA ; Department of Bioengineering, University of Illinois Hospital > Health Sciences System, Chicago, IL, USA.
authors.forename: Randal O
authors.initials: RO
authors.lastname: Dull
authors.affiliation: Department of Anesthesiology, University of Illinois Hospital > Health Sciences System, 835 S. Wolcott Ave (m/c 868), Chicago, IL 60612, USA ; Department of Pharmacology, University of Illinois Hospital > Health Sciences System, Chicago, IL, USA.
authors.forename: Guochang
authors.initials: G
authors.lastname: Hu
authors.affiliation: Department of Anesthesiology, University of Illinois Hospital > Health Sciences System, 835 S. Wolcott Ave (m/c 868), Chicago, IL 60612, USA ; Department of Pharmacology, University of Illinois Hospital > Health Sciences System, Chicago, IL, USA.
authors.forename: Maricela
authors.initials: M
authors.lastname: Castellon
authors.affiliation: Department of Anesthesiology, University of Illinois Hospital > Health Sciences System, 835 S. Wolcott Ave (m/c 868), Chicago, IL 60612, USA.
authors.forename: Andreia Z
authors.initials: AZ
authors.lastname: Chignalia
authors.affiliation: Department of Anesthesiology, University of Illinois Hospital > Health Sciences System, 835 S. Wolcott Ave (m/c 868), Chicago, IL 60612, USA.
authors.forename: Ruben G
authors.initials: RG
authors.lastname: Koshy
authors.affiliation: Department of Anesthesiology, University of Illinois Hospital > Health Sciences System, 835 S. Wolcott Ave (m/c 868), Chicago, IL 60612, USA ; Department of Anesthesiology, Jesse Brown VA Medical Center, Chicago, IL, USA.
authors.forename: E Gina
authors.initials: EG
authors.lastname: Votta-Velis
authors.affiliation: Department of Anesthesiology, Balgrist University Hospital, Zurich, Switzerland.
authors.forename: Alain
authors.initials: A
authors.lastname: Borgeat
authors.affiliation: Department of Anesthesiology, University of Illinois Hospital > Health Sciences System, 835 S. Wolcott Ave (m/c 868), Chicago, IL 60612, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Schwartz
authors.affiliation: Institute of Anesthesiology, University Hospital Zurich, Zurich, Switzerland.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Beck-Schimmer
authors.affiliation: Department of Anesthesiology, University of Illinois Hospital > Health Sciences System, 835 S. Wolcott Ave (m/c 868), Chicago, IL 60612, USA ; Department of Pharmacology, University of Illinois Hospital > Health Sciences System, Chicago, IL, USA ; Department of Bioengineering, University of Illinois Hospital > Health Sciences System, Chicago, IL, USA ; Center for Lung and Vascular Biology, University of Illinois Hospital > Health Sciences System, Chicago, IL, USA.
authors.forename: Richard D
authors.initials: RD
authors.lastname: Minshall
mesh_terms: D055371:Acute Lung Injury / Q000188:drug therapy* / Q000209:etiology; D000577:Amides / Q000494:pharmacology*; D000779:Anesthetics, Local / Q000494:pharmacology*; D000818:Animals; D051242:Caveolin 1 / Q000235:genetics; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000188:drug therapy / Q000473:pathology; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D008070:Lipopolysaccharides / Q000633:toxicity; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010766:Phosphorylation / Q000187:drug effects; D011654:Pulmonary Edema / Q000517:prevention & control; D000077212:Ropivacaine; D015398:Signal Transduction / Q000187:drug effects; D055397:Ventilator-Induced Lung Injury / Q000517:prevention & control; D019061:src-Family Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D000577:Amides; D000779:Anesthetics, Local; D051242:Caveolin 1; D008070:Lipopolysaccharides; D018799:Intercellular Adhesion Molecule-1; D000077212:Ropivacaine; D019061:src-Family Kinases
keywords: ARDS; Acute lung injury; Caveolin-1; Endothelium; Local anesthetics; Src protein tyrosine kinase; Ventilator-induced lung injury
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