Effect of Endotoxin on Primarily Cultured Rat Hepatocyte and Kupffer Cell Mediated Hepatotoxicity

Norio Inoue, Seiji Endo, Rikiya Abe

The Second Department of Surgery, Fukushima Medical School

We studied the role of Kupffer cells in the endotoxin-induced liver injury. Rat hepatocytes and Kupffer cells were primarily cultured, and viability of the hepatocytes were evaluated by MTT assay in vitro. Isolated hepatocytes were cultured in a medium containing different concentrations of lipopolysaccharide (LPS) (0,25, 50, 100 μg/ml) for 24 hours. Viability of hepatocytes was not affected by LPS at any concentration. Isolated Kupffer cells were cultured for 24 hours in medium containing the same concentrations of LPS and no fetal bovine serum, and the medium was harvested. Isolated hepatocytes were added to these media replaced with these medium and cultured for 24 hours. Viability of the hepatocytes cultured in the medium in whick Kupffer cells had been cultured for 24 hours with LPS decreased significantly with increasing LPS concentration. We then performed the same experiment using Kupffer cells isolated from endotoxinemic rats which had been injected LPS (20 μg) 24 hours earlier. The same Kupffer cell-mediated hepatotoxicity was obtained and the toxicity was much stronger than in the case of Kupffer cells isolated from normal rats. Hepatotoxicity of endotoxin in not a direct effect of endotoxin on hepatocytes, but is a Kupffer cell-mediated effect. It is suggested that Kupffer cells might be activated in endotoxinemia.

Key words
Kupffer cell mediated hepatotoxicity, endotoxin induced hepatotoxicity, Kupffer cell, primarily cultured rat hepatocyte, MTT assay

Reprint requests
Norio Inoue The Second Department of Surgery, Fukushima Medical School
1 Hikarigaoka, Fukushima, 960-12 JAPAN

Accepted
May 13, 1992

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