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Volume 272, Number 12, Issue of March 21, 1997 pp. 7582-7585
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

COMMUNICATION:
Bacterial Lipopolysaccharide Plus Interferon-gamma Elicit a Very Fast Inhibition of a Ca2+-dependent Nitric-oxide Synthase Activity in Human Astrocytoma Cells

(Received for publication, November 22, 1996, and in revised form, January 27, 1997)

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

From the ‡ Department of Biology, III University of Rome, 00146 Rome, Italy, the § Institute of Biological Chemistry, University of Verona, 37134 Verona, Italy, and  Department of Biology, University of Rome "Tor Vergata," 00173 Rome, Italy

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-gamma , 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-min treatment of T67 cells with the combination of lipopolysaccharide plus interferon-gamma (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.


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