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To investigate the cellular communication in the liver, nitric oxide (NO) production by sinusoidal cells and hepatocytes by stimulation with cytokines and Kupffer cell-conditioned medium was quantitatively analyzed. NO production by the cells was measured by the Griess reaction, and nitric oxide synthase (iNOS) transcription level by a competitive RT-PCR assay using mutant iNOS mRNA as a standard. NO production and iNOS mRNA transcriptional levels in Kupffer cells were markedly increased by stimulation with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma), and moderately by interleukin-1beta (IL-1beta). NO production by hepatocytes was not significantly enhanced by LPS, but was markedly enhanced by IL-1beta or the combination of tumor-necrosis factor-alpha (TNF-alpha) and IFN-gamma. Hepatocyte NO production and iNOS mRNA levels were markedly enhanced by the LPS-activated Kupffer cell conditioned medium, but these effects were reduced by heat treatment or anti-TNF antibody. Although NG-monomethyl-L-arginine acetate and dexamethasone reduced NO production by the cells, the iNOS mRNA level was reduced by dexamethasone only. Gel-shift assay showed NF-kappaB activation in hepatocytes during this activation. These data reinforce the importance of cellular communication between sinusoidal cells and hepatocytes.
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