Ropivacaine attenuates endotoxin plus hyperinflation-mediated acute lung injury via inhibition of early-onset Src-dependent signaling

Tobias Piegeler, Randal O. Dull, Guochang Hu, Maricela Castellon, Andreia Z. Chignalia, Ruben G. Koshy, E. G. Votta-Velis, Alain Borgeat, David E. Schwartz, Beatrice Beck-Schimmer, Richard D. Minshall

Research output: Contribution to journalArticle

25 Citations (Scopus)

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.

Original languageEnglish (US)
Article number57
JournalBMC Anesthesiology
Volume14
DOIs
StatePublished - Jul 19 2014
Externally publishedYes

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Acute Lung Injury
Endotoxins
Intercellular Adhesion Molecule-1
Caveolin 1
Lung
Phosphorylation
Pulmonary Edema
Artificial Respiration
Blood Vessels
Ventilation
Permeability
Endothelial Cells
Western Blotting
Ventilator-Induced Lung Injury
Extravascular Lung Water
ropivacaine
src-Family Kinases
Water
Tidal Volume
Local Anesthetics

Keywords

  • Acute lung injury
  • ARDS
  • Caveolin-1
  • Endothelium
  • Local anesthetics
  • Src protein tyrosine kinase
  • Ventilator-induced lung injury

ASJC Scopus subject areas

  • Anesthesiology and Pain Medicine

Cite this

Ropivacaine attenuates endotoxin plus hyperinflation-mediated acute lung injury via inhibition of early-onset Src-dependent signaling. / Piegeler, Tobias; Dull, Randal O.; Hu, Guochang; Castellon, Maricela; Chignalia, Andreia Z.; Koshy, Ruben G.; Votta-Velis, E. G.; Borgeat, Alain; Schwartz, David E.; Beck-Schimmer, Beatrice; Minshall, Richard D.

In: BMC Anesthesiology, Vol. 14, 57, 19.07.2014.

Research output: Contribution to journalArticle

Piegeler, T, Dull, RO, Hu, G, Castellon, M, Chignalia, AZ, Koshy, RG, Votta-Velis, EG, Borgeat, A, Schwartz, DE, Beck-Schimmer, B & Minshall, RD 2014, 'Ropivacaine attenuates endotoxin plus hyperinflation-mediated acute lung injury via inhibition of early-onset Src-dependent signaling', BMC Anesthesiology, vol. 14, 57. https://doi.org/10.1186/1471-2253-14-57
Piegeler, Tobias ; Dull, Randal O. ; Hu, Guochang ; Castellon, Maricela ; Chignalia, Andreia Z. ; Koshy, Ruben G. ; Votta-Velis, E. G. ; Borgeat, Alain ; Schwartz, David E. ; Beck-Schimmer, Beatrice ; Minshall, Richard D. / Ropivacaine attenuates endotoxin plus hyperinflation-mediated acute lung injury via inhibition of early-onset Src-dependent signaling. In: BMC Anesthesiology. 2014 ; Vol. 14.
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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.",
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TY - JOUR

T1 - Ropivacaine attenuates endotoxin plus hyperinflation-mediated acute lung injury via inhibition of early-onset Src-dependent signaling

AU - Piegeler, Tobias

AU - Dull, Randal O.

AU - Hu, Guochang

AU - Castellon, Maricela

AU - Chignalia, Andreia Z.

AU - Koshy, Ruben G.

AU - Votta-Velis, E. G.

AU - Borgeat, Alain

AU - Schwartz, David E.

AU - Beck-Schimmer, Beatrice

AU - Minshall, Richard D.

PY - 2014/7/19

Y1 - 2014/7/19

N2 - 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.

AB - 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.

KW - Acute lung injury

KW - ARDS

KW - Caveolin-1

KW - Endothelium

KW - Local anesthetics

KW - Src protein tyrosine kinase

KW - Ventilator-induced lung injury

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