Peroxynitrite causes endothelial cell monolayer barrier dysfunction

James L. Knepler, Loui N. Taher, Mahesh P. Gupta, Carolyn Patterson, Fredrick Pavalko, Michael D. Ober, C. Michael Hart

Research output: Contribution to journalArticle

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Abstract

Nitric oxide (·NO) attenuates hydrogen peroxide (H2O2)-mediated barrier dysfunction in cultured porcine pulmonary artery endothelial cells (PAEC) (Gupta MP, Ober MD, Patterson C, Al-Hassani M, Natarajan V, and Hart, CM. Am J Physiol Lung Cell Mol Physiol 280: L116-L126, 2001). However, ·NO rapidly combines with superoxide (O2-) to form the powerful oxidant peroxynitrite (ONOO-), which we hypothesized would cause PAEC monolayer barrier dysfunction. To test this hypothesis, we treated PAEC with ONOO- (500 μM) or 3-morpholinosydnonimine hydrochloride (SIN-1; 1-500 μM). SIN-1-mediated ONOO- formation was confirmed by monitoring the oxidation of dihydrorhodamine 123 to rhodamine. Both ONOO- and SIN-1 increased albumin clearance (P < 0.05) in the absence of cytotoxicity and altered the architecture of the cytoskeletal proteins actin and β-catenin as detected by immunofluorescent confocal imaging. ONOO--induced barrier dysfunction was partially reversible and was attenuated by cysteine. Both ONOO- and SIN-1 nitrated tyrosine residues, including those on β-catenin and actin, and oxidized proteins in PAEC. The introduction of actin treated with ONOO- into PAEC monolayers via liposomes also resulted in barrier dysfunction. These results indicate that ONOO- directly alters endothelial cytoskeletal proteins, leading to barrier dysfunction.

Original languageEnglish
JournalAmerican Journal of Physiology - Cell Physiology
Volume281
Issue number3 50-3
StatePublished - 2001

Fingerprint

Peroxynitrous Acid
Endothelial cells
Pulmonary Artery
Monolayers
Endothelial Cells
Actins
Catenins
Cytoskeletal Proteins
1-(3'-methoxy-4'-hydroxy-phenyl)-6,7-dihydroxyisochroman
Nitric Oxide
Unilamellar Liposomes
Rhodamines
Cytotoxicity
Oxidants
Liposomes
Superoxides
Hydrogen Peroxide
Cysteine
Tyrosine
Albumins

Keywords

  • Actin
  • Catenin
  • Nitrotyrosine

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology
  • Physiology (medical)

Cite this

Knepler, J. L., Taher, L. N., Gupta, M. P., Patterson, C., Pavalko, F., Ober, M. D., & Hart, C. M. (2001). Peroxynitrite causes endothelial cell monolayer barrier dysfunction. American Journal of Physiology - Cell Physiology, 281(3 50-3).

Peroxynitrite causes endothelial cell monolayer barrier dysfunction. / Knepler, James L.; Taher, Loui N.; Gupta, Mahesh P.; Patterson, Carolyn; Pavalko, Fredrick; Ober, Michael D.; Hart, C. Michael.

In: American Journal of Physiology - Cell Physiology, Vol. 281, No. 3 50-3, 2001.

Research output: Contribution to journalArticle

Knepler, JL, Taher, LN, Gupta, MP, Patterson, C, Pavalko, F, Ober, MD & Hart, CM 2001, 'Peroxynitrite causes endothelial cell monolayer barrier dysfunction', American Journal of Physiology - Cell Physiology, vol. 281, no. 3 50-3.
Knepler, James L. ; Taher, Loui N. ; Gupta, Mahesh P. ; Patterson, Carolyn ; Pavalko, Fredrick ; Ober, Michael D. ; Hart, C. Michael. / Peroxynitrite causes endothelial cell monolayer barrier dysfunction. In: American Journal of Physiology - Cell Physiology. 2001 ; Vol. 281, No. 3 50-3.
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