Prostacyclin post-treatment improves LPS-induced acute lung injury and endothelial barrier recovery via Rap1

Anna A. Birukova, Fanyong Meng, Yufeng Tian, Angelo Meliton, Nicolene Sarich, Lawrence A. Quilliam, Konstantin G. Birukov

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

Protective effects of prostacyclin (PC) or its stable analog beraprost against agonist-induced lung vascular inflammation have been associated with elevation of intracellular cAMP and Rac GTPase signaling which inhibited the RhoA GTPase-dependent pathway of endothelial barrier dysfunction. This study investigated a distinct mechanism of PC-stimulated lung vascular endothelial (EC) barrier recovery and resolution of LPS-induced inflammation mediated by small GTPase Rap1. Efficient barrier recovery was observed in LPS-challenged pulmonary EC after prostacyclin administration even after 15h of initial inflammatory insult and was accompanied by the significant attenuation of p38 MAP kinase and NFκB signaling and decreased production of IL-8 and soluble ICAM1. These effects were reproduced in cells post-treated with 8CPT, a small molecule activator of Rap1-specific nucleotide exchange factor Epac. By contrast, pharmacologic Epac inhibitor, Rap1 knockdown, or knockdown of cell junction-associated Rap1 effector afadin attenuated EC recovery caused by PC or 8CPT post-treatment. The key role of Rap1 in lung barrier restoration was further confirmed in the murine model of LPS-induced acute lung injury. Lung injury was monitored by measurements of bronchoalveolar lavage protein content, cell count, and Evans blue extravasation and live imaging of vascular leak over 6days using a fluorescent tracer. The data showed significant acceleration of lung recovery by PC and 8CPT post-treatment, which was abrogated in Rap1a-/- mice. These results suggest that post-treatment with PC triggers the Epac/Rap1/afadin-dependent mechanism of endothelial barrier restoration and downregulation of p38MAPK and NFκB inflammatory cascades, altogether leading to accelerated lung recovery.

Original languageEnglish (US)
Pages (from-to)778-791
Number of pages14
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Volume1852
Issue number5
DOIs
StatePublished - May 1 2015

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Keywords

  • Cytoskeleton
  • Endothelium
  • Inflammation
  • Lung
  • Permeability

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine

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