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Am J Physiol Renal Physiol 280: F466-F473, 2001;
0363-6127/01 $5.00
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Vol. 280, Issue 3, F466-F473, March 2001

Posttranscriptional regulation of human iNOS by the NO/cGMP pathway

Dolores Pérez-Sala, Eva Cernuda-Morollón, Manuela Díaz-Cazorla, Fernando Rodríguez-Pascual, and Santiago Lamas

Departamento de Estructura y Función de Proteínas, Centro de Investigaciones Biológicas, Madrid, and Instituto "Reina Sofía" de Investigaciones Nefrológicas, CSIC, 28006 Madrid, Spain

Nitric oxide (NO) and cGMP may exert positive or negative effects on inducible NO synthase (iNOS) expression. We have explored the influence of the NO/cGMP pathway on iNOS levels in human mesangial cells. Inhibition of NOS activity during an 8-h stimulation with IL-1beta plus tumor necrosis factor (TNF)-alpha reduced iNOS levels, while NO donors amplified iNOS induction threefold. However, time-course studies revealed a subsequent inhibitory effect of NO donors on iNOS protein and mRNA levels. This suggests that NO may contribute both to iNOS induction and downregulation. Soluble guanylyl cyclase (sGC) activation may be involved in these effects. Inhibition of sGC attenuated IL-1beta /TNF-alpha -elicited iNOS induction and reduced NO-driven amplification. Interestingly, cGMP analogs also modulated iNOS protein and mRNA levels in a biphasic manner. Inhibition of transcription unveiled a negative posttranscriptional modulation of the iNOS transcript by NO and cGMP at late times of induction. Supplementation with 8-bromo-cGMP (8-BrcGMP) reduced iNOS mRNA stability by 50%. These observations evidence a complex feedback regulation of iNOS expression, in which posttranscriptional mechanisms may play an important role.

inflammatory mediators; human mesangial cells; mRNA stability; nitric oxide; inducible nitric oxide synthase; guanosine 3',5'-cyclic monophosphate


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