Murine lung injury caused by Leptospira interrogans glycolipoprotein, a specific Na/K-ATPase inhibitor

Respiratory Research

Mauro Velho de Castro Faria⁴  Hugo Caire de Castro Faria Neto¹  Mauricio Younes-Ibrahim⁴  Patrícia Torres Bozza¹  Verônica Morandi²  Camila Ignácio da Silva³  Emilson Domingos da Silva⁵  Gerson Silva de Lima⁵  Aline Soares Freire²  Ricardo Erthal Santelli²  Flora Magno de Jesus Oliveira¹  Isabel Matos Medeiros de Moraes¹  Adriana Ribeiro Silva¹  Patrícia Burth³  Cassiano Felippe Gonçalves-de-Albuquerque¹ 

[1] Fundação Oswaldo Cruz, Laboratório de Imunofarmacologia, Instituto Oswaldo Cruz, Pavilhão Ozório de Almeida, Av Brasil 4365, Fiocruz, Rio de Janeiro CEP 21040 - 900, RJ, Brazil;Departamento de Química Analítica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil;Departamento de Biologia Celular e Molecular, Instituto de Biologia, Universidade Federal Fluminense, Niterói, RJ, Brazil;Departamento de Medicina Interna, Faculdade de Ciências Médicas, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ, Brazil;Fundação Oswaldo Cruz, Instituto de Tecnologia em Imunobiológicos-BIO Manguinhos, Departamento de Reativos Para Diagnóstico, Av Brasil 4365 Fiocruz, Rio de Janeiro CEP 21040 - 900, RJ, Brazil

关键词: MAPK p38;    Ouabain;    Toll-like receptors;    Lung injury;    Na/K-ATPase;    Leptospiral GLP;   

Others  :  1146491
DOI  :  10.1186/s12931-014-0093-2

 received in 2014-03-19, accepted in 2014-07-31,  发布年份 2014

【 摘 要 】

Background

Leptospiral glycolipoprotein (GLP) is a potent and specific Na/K-ATPase inhibitor. Severe pulmonary form of leptospirosis is characterized by edema, inflammation and intra-alveolar hemorrhage having a dismal prognosis. Resolution of edema and inflammation determines the outcome of lung injury. Na/K-ATPase activity is responsible for edema clearance. This enzyme works as a cell receptor that triggers activation of mitogen-activated protein kinase (MAPK) intracellular signaling pathway. Therefore, injection of GLP into lungs induces injury by triggering inflammation.

Methods

We injected GLP and ouabain, into mice lungs and compared their effects. Bronchoalveolar lavage fluid (BALF) was collected for cell and lipid body counting and measurement of protein and lipid mediators (PGE₂ and LTB₄). The levels of the IL-6, TNFα, IL-1B and MIP-1α were also quantified. Lung images illustrate the injury and whole-body plethysmography was performed to assay lung function. We used Toll-like receptor 4 (TLR4) knockout mice to evaluate leptospiral GLP-induced lung injury. Na/K-ATPase activity was determined in lung cells by nonradioactive rubidium incorporation. We analyzed MAPK p38 activation in lung and in epithelial and endothelial cells.

Results

Leptospiral GLP and ouabain induced lung edema, cell migration and activation, production of lipid mediators and cytokines and hemorrhage. They induced lung function alterations and inhibited rubidium incorporation. Using TLR4 knockout mice, we showed that the GLP action was not dependent on TLR4 activation. GLP activated of p38 and enhanced cytokine production in cell cultures which was reversed by a selective p38 inhibitor.

Conclusions

GLP and ouabain induced lung injury, as evidenced by increased lung inflammation and hemorrhage. To our knowledge, this is the first report showing GLP induces lung injury. GLP and ouabain are Na/K-ATPase targets, triggering intracellular signaling pathways. We showed p38 activation by GLP-induced lung injury, which was may be linked to Na/K-ATPase inhibition. Lung inflammation induced by GLP was not dependent on TLR4 activation.

【 授权许可】

   
2014 Gonçalves-de-Albuquerque et al.; licensee BioMed Central Ltd.

【 图 表 】

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