Cytoprotective-selective activated protein C attenuates Pseudomonas aeruginosa-induced lung injury in mice.
Inhibition of the small GTPase RhoA attenuates the development of pulmonary edema and restores positive alveolar fluid clearance in a murine model of Pseudomonas aeruginosa pneumonia. Activated protein C (aPC) blocks the development of an unfavorably low ratio of small GTPase Rac1/RhoA activity in lung endothelium through endothelial protein C receptor (EPCR)/protease-activated receptor-1 (PAR-1)-dependent signaling mechanisms that include transactivating the sphingosine-1-phosphate (S1P) pathway. However, whether aPC's cytoprotective effects can attenuate the development of pulmonary edema and death associated with P. aeruginosa pneumonia in mice remains unknown. Thus, we determined whether the normalization of a depressed ratio of activated Rac1/RhoA by aPC would attenuate the P. aeruginosa-mediated increase in protein permeability across lung endothelial and alveolar epithelial barriers. Pretreatment with aPC significantly reduced P. aeruginosa-induced increases in paracellular permeability across pulmonary endothelial cell and alveolar epithelial monolayers via an inhibition of RhoA activation and a promotion of Rac1 activation that required the EPCR-PAR-1 and S1P pathways. Furthermore, pretreatment with aPC attenuated the development of pulmonary edema in a murine model of P. aeruginosa pneumonia. Finally, a cytoprotective-selective aPC mutant, aPC-5A, which lacks most of aPC's anticoagulant activity, reproduced the protective effect of wild-type aPC by attenuating the development of pulmonary edema and decreasing mortality in a murine model of P. aeruginosa pneumonia. Taken together, these results demonstrate a critical role for the cytoprotective activities of aPC in attenuating P. aeruginosa-induced lung vascular permeability and mortality, suggesting that cytoprotective-selective aPC-5A with diminished bleeding risks could attenuate the lung damage caused by P. aeruginosa in critically ill patients.
Cardiovascular Research Institute and Institute of Molecular Medicine, Department of Anesthesia, University of California at San Francisco, San Francisco, California, USA.
Nastasha
N
Bir
Mathieu
M
Lafargue
Marybeth
M
Howard
Arnaud
A
Goolaerts
Jeremie
J
Roux
Michel
M
Carles
Mitchell J
MJ
Cohen
Karen E
KE
Iles
José A
JA
Fernández
John H
JH
Griffin
Jean-Francois
JF
Pittet
D000818:Animals; D002417:Cattle; D002460:Cell Line; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000166:cytology; D006801:Humans; D008168:Lung / Q000382:microbiology*; D055370:Lung Injury / Q000382:microbiology*; D051379:Mice; D011486:Protein C / Q000378:metabolism*; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D011654:Pulmonary Edema / Q000378:metabolism; D051381:Rats; D020830:rac1 GTP-Binding Protein / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
D011486:Protein C; C107953:RAC1 protein, human; C066737:RHOA protein, human; C526380:RhoA protein, mouse; D020830:rac1 GTP-Binding Protein; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein
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