Hemin causes lung microvascular endothelial barrier dysfunction by necroptotic cell death

Sunit Singla, Justin R. Sysol, Benjamin Dille, Nicole Jones, Jiwang Chen, Roberto F. Machado

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Hemin, the oxidized prosthetic moiety of hemoglobin, has been implicated in the pathogenesis of acute chest syndrome in patients with sickle cell disease by virtue of its endothelial-activating properties. In this study, we examined whether hemin can cause lung microvascular endothelial barrier dysfunction. By assessing transendothelial resistance using electrical cell impedance sensing, and by directly measuring trans-monolayer fluorescein isothiocyanate-dextran flux, we found that hemin does cause endothelial barrier dysfunction in a concentration-dependent manner. Pretreatment with either a Toll-like receptor 4 inhibitor, TAK-242, or an antioxidant, N-acetylcysteine, abrogated this effect. Increased monolayer permeability was found to be associated with programmed cell death by necroptosis, as evidenced by Trypan blue staining, terminal deoxynucleotidyl transferase dUTP nick-end labeling assay, Western blotting for activated forms of key effectors of cell death pathways, and studies utilizing specific inhibitors of necroptosis and apoptosis. Further studies examining the role of endothelial cell necroptosis in promoting noncardiogenic pulmonary edema during acute chest syndrome are warranted and may open a new avenue of potential treatments for this devastating disease.

Original languageEnglish (US)
Pages (from-to)307-314
Number of pages8
JournalAmerican journal of respiratory cell and molecular biology
Volume57
Issue number3
DOIs
StatePublished - Sep 2017
Externally publishedYes

Keywords

  • Acute chest syndrome
  • Endothelial cell
  • Hemin
  • Necroptosis
  • Sickle cell disease

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

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