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Research in immunology, v.149 no.2, 1998년, pp.139 - 150
Sinha, B. (Corresponding author.) , Eigler, A. , Baumann, K.H. , Greten, T.F. , Moeller, J. , Endres, S.
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SummaryNitric oxide (NO) and tumour necrosis factor (TNF) are essential mediators in a number of biological processes, including the immune response. TNF stimulates NO production via expression of inducible NO synthase (iNOS), with L-arginine being the only substrate. Previously, we demonstrated tha...
Sachant que la synthèse de TNF (facteur nécrotique tumoral), induite par le LPS (lipopolyoside), est inhibée par NO, nous avons cherché à savoir si une réduction de l'activité de TNF est reflétée au niveau de l'ARNm de TNF dans la lignée RAW 264.7 de macrophages murins. L'ARNm a été quantifié par la méthode Northern avec une sonde marquée par α 32P-dCTP. Les cellules stimulées par 10 μg de LPS/ml en l'absence de L-arginine (afin de prévenir la formation de NO endogène) contiennent plus d'ARNm que les cellules additionnées de 1 mM de L-arginine en contiennent, 14 et 20 h après stimulation; aucune différence n'est observée après 4 h. Une telle cinétique est compatible avec l'implication de la NO synthase inductible. La demi-vie de l'ARNm en présence de NO est approximativement la moitié de celle notée en l'absence de L-arginine (41 min au lieu de 77 min). La L-citrulline, à 1 mM, qui est recyclée en L-arginine dans les cellules, compense entièrement la réduction de l'activité du TNF, et ces résultats suggèrent que le NO endogène régule l'ARNm du TNF surtout en réduisant la demi-vie. En outre, une bande supplémentaire d'environ 1,4 kb (hybridation avec TNF) est toujours observé chez les cellules non stimulées. Cela peut correspondre à un ARNm spécifiquement hydrolysé dans la région riche en AU, reflétant possiblement un autre point de régulation de l'expression du TNF.
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