Roles of il-1 and tnf-[alpha] in endotoxin-induced activation of nitric oxide synthase in cultured rat brain cells. Romero, Luz I., Jeffrey B. Tatro, Jodie A. Field, and Seymour Reichlin. Division of Endocrinology, Diabetes, Metabolism and Molecular Medicine, Tufts University School of Medicine and New England Medical Center Hospitals, Boston, MA 02111
APStracts 2:0267R, 1995.
In astrocytes and microglia, bacterial lipopolysaccharide (LPS) stimulates production and release of interleukin-1[beta] (IL -1[beta]), tumor necrosis factor-[alpha] (TNF[alpha]) and nitric oxide (NO). Although IL-1[beta] and TNF[alpha] are themselves capable of inducing NO synthase (NOS) in glia, the specific factors mediating LPS induction of NOS in brain have not been identified. To determine whether LPS induction of NOS in brain cells is mediated by IL-1 or TNF[alpha], acting alone or in concert, the effects of IL-1 receptor antagonist (IL-1Ra) and of TNF soluble receptor (TNFsRp55), presented individually and in combination, on LPS-induced NOS activity were tested. In glia-enriched mixed primary cultures of neonatal rat telencephalic cells, LPS (0.1-100 ng/ml), IL-1[beta] (0.01-10 nM), and TNF[alpha] (0.1-100 nM) each concentration-dependently stimulated accumulation of nitrite, an indicator of NO production. Induction of nitrite accumulation by LPS and by IL1 was blocked by Nw-nitro-L -arginine methyl ester (NAME) and Nw-monomethyl-L-arginine (LNMA), indicating that it was mediated by nitric oxide synthase. TNF[alpha] alone induced NO production weakly as compared with IL-1, but combined submaximal concentrations of IL-1[beta] (1 nM) and TNF[alpha] (10 nM) induced NOS synergistically. Furthermore, TNFsRp55 and IL-1Ra each produced a dose-dependent partial inhibition of the NO response to LPS, and the effect of TNFsRp55 was equal to or greater than that of IL-1Ra. TNFsRp55 and IL1-Ra in combination were not significantly more effective than TNFsRp55 alone. The results indicate that LPS induction of NOS activity in brain cells is mediated in part by both IL-1[beta] and TNF[alpha].

Received 6 March 1995; accepted in final form 16 July 1995.
APS Manuscript Number R145-5.
Article publication pending Am. J. Physiol. (Regulatory Integrative
Comp. Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 31 October 95