Bacterial lipopolysaccharide plus interferon-γ elicit a very fast inhibition of a Ca2+-dependent nitric-oxide synthase activity in human astrocytoma cells

Marco Colasanti, Elisabetta Cavalieri, Tiziana Persichini, Vincenzo Mollace, Sofia Mariotto, Hisanori Suzuki, Giuliana M. Lauro

Research output: Contribution to journalArticle

Abstract

Previous results indicate that induction of inducible nitric-oxide synthase (iNOS) expression may be kept suppressed by the endogenous NO level as produced by a constitutive NOS (cNOS) enzyme. In cell types possessing both cNOS and iNOS, this may represent an evident paradox. Here, we report that lipopolysaccharide and interferon-γ, which are able to strongly induce iNOS in astrocytoma cells, can rapidly inhibit the NO production generated by the constitutive NOS isoform, thus obtaining the best conditions for iNOS induction and resolving the apparent paradox. In fact, a 30-rain treatment of T67 cells with the combination of lipopolysaccharide plus interferon-γ (MIX) strongly inhibits the cNOS activity, as determined by measuring [3H]citrulline production. In addition, the effect of MIX is also observed by measuring nitrite, the stable breakdown product of NO: a 30-min pretreatment of T67 cells with MIX is able to reduce significantly the N- methyl-D-aspartate-induced nitrite production. Finally, using reverse transcriptase-polymerase chain reaction, we have observed that a 30-min treatment of T67 cells with MIX does not affect expression of mRNA coding for the neuronal NOS-I isoform. These results suggest the novel concept of a possible role of a cNOS isoform in astrocytes as a control function on iNOS induction.

Original languageEnglish
Pages (from-to)7582-7585
Number of pages4
JournalJournal of Biological Chemistry
Volume272
Issue number12
DOIs
Publication statusPublished - 1997

ASJC Scopus subject areas

  • Biochemistry

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