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Regulation of ICAM‐1/CD54 expression on human endothelial cells by hydrogen peroxide involves inducible NO synthase
Author(s) -
Zadeh Majid Sadeghi,
Kolb JeanPierre,
Geromin Daniela,
D'Anna Rossella,
Boulmerka Amira,
Marconi Anthony,
Dugas Bernard,
Marsac Cécile,
D'Alessio Patrizia
Publication year - 2000
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.67.3.327
Subject(s) - nitric oxide synthase , biology , microbiology and biotechnology , nitric oxide , western blot , icam 1 , citrulline , cell adhesion molecule , hydrogen peroxide , vcam 1 , biochemistry , intracellular , arginine , endocrinology , amino acid , gene
Expression of the inducible isoform of nitric oxide synthase (iNOS) is stimulated by cytokines in human epithelial cells. This work indicates that incubation of human umbilical cord endothelial cells with combinations of interleukin‐1β, tumor necrosis factor α, and interferon‐γ stimulated the synthesis of iNOS mRNA, as detected by reverse transcriptase‐polymerase chain reaction. It is important to note that 50, 100, and 200 μM hydrogen peroxide was able to stimulate iNOS directly. Furthermore, 100 μM H 2 O 2 enhanced synthesis of the oxidation products, nitrite (NO) and nitrate (NO) at 12 and 36 h. iNOS protein, detected by Western blot analysis, as well as L‐citrulline levels, were also increased. When endothelial cell monolayers were incubated for 1 h with 100 μM H 2 O 2 and subsequently with cytokines, iNOS mRNA was further augmented. Under the same conditions, we regularly observed an inhibition (25%) of intercellular adhesion molecule‐1 (ICAM‐1/CD54) expression. The latter was reversed when the NOS inhibitor N G ‐monomethyl‐L‐arginine was added, as shown by flow cytometry. These data suggest a specific effect of endogenous hydroperoxides on the biosynthesis and processing of the human endothelial iNOS isoform. We propose that H 2 O 2 induces a temporary NO‐dependent modulation of adhesion molecule expression to limit the tissue destruction that accompanies the vascular recruitment of leukocytes. J. Leukoc. Biol. 67: 327–334; 2000.