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Fas ligand expression and apoptosis in primary rat hepatocytes induced by lipopolysaccharide
1Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Objective:
To study hepatocyte apoptosis induced by lipopolysaccharide (LPS) directly and indirectly, and to elucidate the mechanisms of liver damage in endotoxemia.
Methods:
Rat hepatocytes were isolated using collagenase perfusion, and cultured in RPMI 1640 medium. After 24h or 48 h of LPS treatment at various concentrations (1, 5, 10 mug/ml), membrane-bound Fas ligand (mFasL) expression in hepatocytes was determined by immunocytochemistry, and apoptosis was detected by TUNEL. In another set of experiments it was examined whether LPS-treated hepatocytes and its supernatants can stimulate apoptosis in LPS-untreated hepatocytes.
Results:
LPS markedly stimulated mFasL expression and apoptosis in hepatocytes in a dose and time (24-48 h) dependent manner. In the co-culture system LPS-treated hepatocytes significantly induced LPS-untreated hepatocyte apoptosis. In contrast, there was no apoptotic cells observed in the supernatant stimulation system.
Conclusion:
LPS not only directly causes hepatocyte apoptosis, but also indirectly induces apoptosis of LPS-untreated hepatocytes by way of stimulating mFasL expression in hepatocytes.
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