Lead stimulates intercellular signalling between hepatocytes and Kupffer cells.
The role of intercellular signalling between liver cells in lead (Pb)(1)-induced liver toxicity was investigated in cocultures of freshly isolated and cultured rat hepatocytes and Kupffer cells. The Kupffer cells (seeded onto culture dish inserts), the hepatocytes or the two in cocultures were exposed to Pb acetate (2-50 microM) in combination with lipopolysaccharide (0.1-1000 ng/ml). In hepatocyte cultures, the combined Pb/lipopolysaccharide treatment induced no significant increase in the release of the proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) whereas in Kupffer cell cultures and in cocultures, at low lipopolysaccharide levels (0.1 and 1 ng/ml), TNF-alpha release was synergistically increased (up to 30-fold) when compared to lipopolysaccharide exposure alone. This stimulation of Kupffer cell-derived TNF-alpha release was specific for Pb or not detectable with mercury and cadmium. As a response to the Pb/lipopolysaccharide induced release of TNF-alpha, the cocultured hepatocytes increased their nitric oxide (NO) content sixfold when compared with lipopolysaccharide alone and downregulated the negatively regulated acute phase protein albumin. This downregulation was also detectable without lipopolysaccharide and without TNF-alpha release, indicating that Pb induces additional thus far unidentified Kupffer cell-derived factors, which interact with the cocultured hepatocytes. At the time of TNF-alpha release, the viability of the hepatocytes and the Kupffer cells was not affected. However, after a 48-h treatment period, Pb induced a Kupffer cell specific toxicity without affecting the hepatocytes. Loss of hepatocyte viability after lipopolysaccharide/Pb stimulation was only detectable in the presence of cocultured Kupffer cells together with human-derived granulocytes. It is concluded that Pb stimulates intercellular signalling between Kupffer cells and hepatocytes which is synergistically enhanced in the presence of low lipopolysaccharide levels. The released Kupffer cell-derived signals (e.g. cytokines) promotes most likely proteolytic hepatocyte killing in combination with a direct cellular interaction between the granulocytes and the hepatocytes.
Institute of Toxicology, Swiss Federal Institute of Technology Zürich, Schorenstr. 16, CH-8603, Schwerzenbach, Switzerland.
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N
Milosevic
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P
Maier
D000085:Acetates / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D007611:Aprotinin / Q000494:pharmacology; D002104:Cadmium / Q000494:pharmacology; D002450:Cell Communication / Q000187:drug effects* / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D006098:Granulocytes / Q000166:cytology / Q000187:drug effects; D007728:Kupffer Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007854:Lead / Q000378:metabolism / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008628:Mercury / Q000494:pharmacology; D009569:Nitric Oxide / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000187:drug effects / Q000378:metabolism / Q000494:pharmacology
D000085:Acetates; D017077:Culture Media, Conditioned; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D002104:Cadmium; D007854:Lead; D009569:Nitric Oxide; D007611:Aprotinin; C028031:cadmium acetate; D008628:Mercury; C029823:mercuric acetate