Rho inhibition decreases TNF-induced endothelial MAPK activation and monolayer permeability

Fiemu E. Nwariaku, Patricia Rothenbach, Zijuan Liu, Xudong Zhu, Richard H. Turnage, Lance S. Terada

Research output: Contribution to journalArticle

65 Scopus citations

Abstract

Our laboratory previously demonstrated that MAPK activation is an important signal during cytokine-induced endothelial permeability (Nwariaku FE, Liu Z, Terada L, Duffy S, Sarosi G, and Turnage R. Shock 18: 82-85, 2002). Because GTP-binding proteins have been implicated in MAPK activation, we now hypothesize that the GTP-binding protein Rho is a mediator of TNF-induced MAPK activation and increased endothelial permeability. Transmonolayer permeability was assessed in human lung microvascular cells by measuring transmonolayer electrical resistance. MAPK activity was assessed by using a phospho-specific immunoprecipitation kinase assay and by comparing Western blots for phospho-MAPK with total MAPK. MAPK inhibitors used were SB-202190 and PD-098059, whereas Clostridium botulinum C3 transferase was used as a Rho inactivator. Rho-associated coiled-coil kinase was inhibited with Y-27632. TNF increased pulmonary endothelial permeability in vitro and caused a rapid, sustained increase in endothelial p38 and extracellular signal-regulated kinase MAPK activity. Inhibition of p38 and extracellular signal-regulated kinase MAPK with SB-202190 and PD-098059, respectively, decreased TNF-induced endothelial permeability. C3 transferase attenuated TNF-induced MAPK activation and blocked TNF-induced endothelial permeability. Finally, inhibition of Rho-associated coiled-coil kinase with Y-27632 prevented both MAPK activation and TNF-induced decreases in transmonolayer resistance. Rho acts upstream of mitogen-activated protein kinases in mediating TNF-induced pulmonary endothelial leak.

Original languageEnglish (US)
Pages (from-to)1889-1895
Number of pages7
JournalJournal of applied physiology
Volume95
Issue number5
DOIs
StatePublished - Nov 2003

Keywords

  • Cell signaling
  • Endothelium
  • Extracellular signal-regulated kinase
  • Guanosine 5′-triphosphate-binding proteins; cytokines
  • Mitogen-activated protein kinase
  • P38
  • Tumor necrosis factor

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Fingerprint Dive into the research topics of 'Rho inhibition decreases TNF-induced endothelial MAPK activation and monolayer permeability'. Together they form a unique fingerprint.

  • Cite this