cGMP increases antioxidant function and attenuates oxidant cell death in mouse lung microvascular endothelial cells by a protein kinase G-dependent mechanism

R. Scott Stephens, Otgonchimeg Rentsendorj, Laura E. Servinsky, Aigul Moldobaeva, Rachel Damico, David B. Pearse

Research output: Contribution to journalArticle

Abstract

Increasing evidence suggests that endothelial cytotoxicity from reactive oxygen species (ROS) contributes to the pathogenesis of acute lung injury. Treatments designed to increase intracellular cGMP attenuate ROS-mediated apoptosis and necrosis in several cell types, but the mechanisms are not understood, and the effect of cGMP on pulmonary endothelial cell death remains controversial. In the current study, increasing intracellular cGMP by either 8pCPT-cGMP (50 μM) or atrial natriuretic peptide (10 nM) significantly attenuated cell death in H2O2-challenged mouse lung microvascular (MLMVEC) monolayers. 8pCPT-cGMP also decreased perfusate LDH release in isolated mouse lungs exposed to H2O2 or ischemia-reperfusion. The protective effect of increasing cGMP in MLMVECs was accompanied by enhanced endothelial H2O2 scavenging (measured by H2O2 electrode) and decreased intracellular ROS concentration (measured by 2′,7′-dichlorofluorescin fluorescence) as well as decreased phosphorylation of p38 MAPK and Akt. The cGMP-mediated cytoprotection and increased H2O2 scavenging required >2 h of 8pCPT-cGMP incubation in wild-type MLMVEC and were absent in MLMVEC from protein kinase G (PKGI)-/- mice suggesting a PKG I-mediated effect on gene regulation. Catalase and glutathione peroxidase 1 (Gpx-1) protein were increased by cGMP in wild-type but not PKGI-/-MLMVEC monolayers. Both the cGMP-mediated increases in antioxidant proteins and H2O2 scavenging were prevented by inhibition of translation with cycloheximide. 8pCPT-cGMP had minimal effects on catalase and Gpx-1 mRNA. We conclude that cGMP, through PKGI, attenuated H 2O2-induced cytotoxicity in MLMVEC by increasing catalase and Gpx-1 expression through an unknown posttranscriptional effect.

Original languageEnglish (US)
Pages (from-to)L323-L333
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume299
Issue number3
DOIs
StatePublished - Sep 2010

Keywords

  • Apoptosis
  • Catalase
  • Glutathione peroxidase
  • Hydrogen peroxide
  • Necrosis

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology

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