Respiratory syncytial virus inhibits lung epithelial Na+ channels by up-regulating inducible nitric-oxide synthase

Academic Article


  • Respiratory syncytial virus (RSV) infection has been shown to reduce Na+-driven alveolar fluid clearance in BALB/c mice in vivo. To investigate the cellular mechanisms by which RSV inhibits amiloride-sensitive epithelial Na+ channels (ENaC), the main pathways through which Na+ ions enter lung epithelial cells, we infected human Clara-like lung (H441) cells with RSV that expresses green fluorescent protein (rRA2). 3-6 days later patch clamp recordings showed that infected cells (i.e. cells expressing green fluorescence; GFP(+)) had significantly lower whole-cell amiloride-sensitive currents and single channel activity (NPo) as compared with non-infected (GFP(-)), non-inoculated, or cells infected with UV-inactivated RSV. Both α and β ENaC mRNA levels were significantly reduced in GFP(+) cells as measured by real-time reverse transcription-PCR. Infection with RSV increased expression of the inducible nitric-oxide synthase (iNOS) and nitrite concentration in the culture medium; nuclear translocation of NF-κB p65 subunit and NF-κB activation were also up-regulated. iNOS up-regulation in GFP(+) cells was prevented by knocking down IκB kinase γ before infection. Furthermore, pretreatment of H441 cells with the specific iNOS inhibitor 1400W (1μM) resulted in a doubling of the amiloride-sensitive Na+ current in GFP(+) cells. Additionally, preincubation of H441 cells with A77-1726 (20μM), a de novo UTP synthesis inhibitor, and 1400W completely reversed the RSV inhibition of amiloride-sensitive currents in GFP(+) cells. Thus, both UTP- and iNOS-generated reactive species contribute to ENaC down-regulation in RSV-infected airway epithelial cells. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.
  • Published In

    Digital Object Identifier (doi)

    Author List

  • Song W; Liu G; Bosworth CA; Walker JR; Megaw GA; Lazrak A; Abraham E; Sullender WM; Matalon S
  • Start Page

  • 7294
  • End Page

  • 7306
  • Volume

  • 284
  • Issue

  • 11