Am J Physiol Cell Physiol AJP: Endocrinology and Metabolism
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Am J Physiol Cell Physiol 275: C208-C215, 1998;
0363-6143/98 $5.00
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Vol. 275, Issue 1, C208-C215, July 1998

Tyrosine kinase inhibitors and antioxidants modulate NF-kappa B and NOS-II induction in retinal epithelial cells

Violaine Faure, Yves Courtois, and Olivier Goureau

Développement, Vieillissement, et Pathologie de la Rétine, Unité 450 Institut National de la Santé et de la Recherche Médicale, Association Claude Bernard, 75016 Paris, France

Bovine retinal pigmented epithelial (RPE) cells express an inducible nitric oxide synthase (NOS-II) after activation with interferon-gamma (IFN-gamma ) and lipopolysaccharide (LPS). Experiments were performed to investigate the effects of tyrosine kinase inhibitors (genistein and herbimycin A) and antioxidants [pyrrolidine dithiocarbamate (PDTC) and butyl hydroxyanisol] on NOS-II induction. The LPS-IFN-gamma -induced nitrite release was inhibited in a concentration-dependent manner by these compounds. Analysis by Northern blot showed that this inhibitory effect correlated with a decrease in NOS-II mRNA accumulation. Analysis by electrophoretic mobility shift assay of the activation of the transcription factor nuclear factor-kappa B (NF-kappa B) involved in NOS-II induction demonstrated that LPS alone or combined with IFN-gamma induced NF-kappa B binding. NF-kappa B activation was not changed by the presence of tyrosine kinase inhibitors but was totally prevented by PDTC pretreatment. Immunocytochemistry experiments confirmed the reduction of the nuclear translocation of NF-kappa B only by PDTC. Our results demonstrated the existence in retinal pigmented epithelial cells of different intracellular signaling pathways in NOS-II induction, since tyrosine kinase inhibitors blocked NOS-II mRNA accumulation without inhibiting NF-kappa B activation. Furthermore, the LPS-IFN-gamma -induced NOS-II mRNA accumulation was sensitive to cycloheximide, suggesting that, in addition to NF-kappa B, transcriptional factors that require new protein synthesis are involved in NOS-II induction.

interferon-gamma ; lipopolysaccharide; nitric oxide; transcription factor


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