Inhibition of Pyk2 blocks lung inflammation and injury in a mouse model of acute lung injury

Duan Yingli, Learoyd Jonathan, Meliton, Leff, Zhu, Zhu Xiangdong

Description

Proline-rich tyrosine kinase 2 (Pyk2) is essential in neutrophil degranulation and chemotaxis in vitro. However, its effect on the process of lung inflammation and edema formation during LPS induced acute lung injury (ALI) remains unknown. The goal of the present study was to determine the effect of inhibiting Pyk2 on LPS-induced acute lung inflammation and injury in vivo. Methods C57BL6 mice were given either 10 mg/kg LPS or saline intratracheally. Inhibition of Pyk2 was effected by intraperitoneal administration TAT-Pyk2-CT 1 h before challenge. Bronchoalveolar lavage analysis of cell counts, lung histology and protein concentration in BAL were analyzed at 18 h after LPS treatment. KC and MIP-2 concentrations in BAL were measured by a mouse cytokine multiplex kit. The static lung compliance was determined by pressure-volume curve using a computer-controlled small animal ventilator. The extravasated Evans blue concentration in lung homogenate was determined spectrophotometrically. Results Intratracheal instillation of LPS induced significant neutrophil infiltration into the lung interstitium and alveolar space, which was attenuated by pre-treatment with TAT-Pyk2-CT. TAT-Pyk2-CT pretreatment also attenuated 1) myeloperoxidase content in lung tissues, 2) vascular leakage as measured by Evans blue dye extravasation in the lungs and the increase in protein concentration in bronchoalveolar lavage, and 3) the decrease in lung compliance. In each paradigm, treatment with control protein TAT-GFP had no blocking effect. By contrast, production of neutrophil chemokines MIP-2 and keratinocyte-derived chemokine in the bronchoalveolar lavage was not reduced by TAT-Pyk2-CT. Western blot analysis confirmed that tyrosine phosphorylation of Pyk2 in LPS-challenged lungs was reduced to control levels by TAT-Pyk2-CT pretreatment. Conclusions These results suggest that Pyk2 plays an important role in the development of acute lung injury in mice and that pharmacological inhibition of Pyk2 might provide a potential therapeutic strategy in the pretreatment for patients at imminent risk of developing acute lung injury.

Sujets

• Inflammation
• Lipopolysaccharide
• Lung
• Neutrophil granulocyte
• PTK2B

Informations

• Publié par: biomed
• Publié le: 01 janvier 2012
• Nombre de lectures: 7
• Langue: English
• Poids de l'ouvrage: 1 Mo

Extrait

Duanet al.Respiratory Research2012,13:4 http://respiratoryresearch.com/content/13/1/4

R E S E A R C HOpen Access Inhibition of Pyk2 blocks lung inflammation and injury in a mouse model of acute lung injury 1 11 1,21* Yingli Duan , Jonathan Learoyd , Angelo Y Meliton , Alan R Leffand Xiangdong Zhu

Abstract Background:Prolinerich tyrosine kinase 2 (Pyk2) is essential in neutrophil degranulation and chemotaxis in vitro. However, its effect on the process of lung inflammation and edema formation during LPS induced acute lung injury (ALI) remains unknown. The goal of the present study was to determine the effect of inhibiting Pyk2 on LPS induced acute lung inflammation and injury in vivo. Methods:C57BL6 mice were given either 10 mg/kg LPS or saline intratracheally. Inhibition of Pyk2 was effected by intraperitoneal administration TATPyk2CT 1 h before challenge. Bronchoalveolar lavage analysis of cell counts, lung histology and protein concentration in BAL were analyzed at 18 h after LPS treatment. KC and MIP2 concentrations in BAL were measured by a mouse cytokine multiplex kit. The static lung compliance was determined by pressurevolume curve using a computercontrolled small animal ventilator. The extravasated Evans blue concentration in lung homogenate was determined spectrophotometrically. Results:Intratracheal instillation of LPS induced significant neutrophil infiltration into the lung interstitium and alveolar space, which was attenuated by pretreatment with TATPyk2CT. TATPyk2CT pretreatment also attenuated 1) myeloperoxidase content in lung tissues, 2) vascular leakage as measured by Evans blue dye extravasation in the lungs and the increase in protein concentration in bronchoalveolar lavage, and 3) the decrease in lung compliance. In each paradigm, treatment with control protein TATGFP had no blocking effect. By contrast, production of neutrophil chemokines MIP2 and keratinocytederived chemokine in the bronchoalveolar lavage was not reduced by TATPyk2CT. Western blot analysis confirmed that tyrosine phosphorylation of Pyk2 in LPS challenged lungs was reduced to control levels by TATPyk2CT pretreatment. Conclusions:These results suggest that Pyk2 plays an important role in the development of acute lung injury in mice and that pharmacological inhibition of Pyk2 might provide a potential therapeutic strategy in the pretreatment for patients at imminent risk of developing acute lung injury. Keywords:inflammation, lipopolysaccharide, lung, neutrophils, Pyk2

Background Acute lung injury (ALI), which may progress to Acute Respiratory Distress Syndrome (ARDS), is associated with high morbidity and mortality in critically ill patients [1,2]. Despite intense research and multiple diverse therapeutic trials, there still are few effective measures for prevention or treatment of ARDS. ARDS is a frequent complication that emerges in patients hav ing sepsis. Lipopolysaccharides (LPS) components of

* Correspondence: [e-mail] 1 Section of Pulmonary and Critical Care Medicine, Department of Medicine, The University of Chicago, Chicago, IL 60637 Full list of author information is available at the end of the article

endotoxin are responsible for the enhanced inflamma tory response of ALI and ARDS [3]. The LPS induced mouse model of ALI is associated with increased neu trophilic lung inflammation and endothelial barrier dys function [46]. Intranasal instillation of LPS stimulates airway epithelial cells to release proinflammatory cyto kines and chemotactic factors, which causes subsequent neutrophilic infiltration and ultimately results in lung tissue injury [7]. This study was designed to determine whether inhibition of the protein tyrosine kinase Pyk2, which mediates a wide variety of cellular activities including cell migration [8], blocks neutrophil infiltra tion and lung injury induced by LPS in mice.

© 2012 Duan et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.