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-Induced Hepatic Parenchymal Cell Apoptosis and Neutrophil-Mediated Necrosis in a Murine Endotoxin Shock Model1

*
Department of Pharmacology, Pharmacia & Upjohn, Inc., Kalamazoo, MI 49007; and
Department of Pathology, University of Texas Health Science Center, Houston, TX 77030
Endotoxin (ET)-induced liver failure is characterized by
parenchymal cell apoptosis and inflammation leading to liver cell
necrosis. Members of the caspase family have been implicated in the
signal transduction pathway of apoptosis. The aim of this study was to
characterize ET-induced hepatic caspase activation and apoptosis and to
investigate their effect on neutrophil-mediated liver injury. Treatment
of C3Heb/FeJ mice with 700 mg/kg galactosamine (Gal) and 100 µg/kg
Salmonella abortus equi ET increased caspase 3-like
protease activity (Asp-Val-Glu-Asp-substrate) by 1730 ± 140% at
6 h. There was a parallel enhancement of apoptosis (assessed by
DNA fragmentation ELISA and terminal deoxynucleotidyl
transferase-mediated dUTP nick end labeling assay). In contrast,
activity of caspase 1 (IL-1ß-converting enzyme)-like proteases
(Tyr-Val-Ala-Asp-substrate) did not change throughout the experiment.
Caspase 3-like protease activity and apoptosis was not induced by
Gal/ET in ET-resistant mice (C3H/HeJ). Furthermore, only murine TNF-
but not IL-1
ß increased caspase activity and apoptosis. Gal/ET
caused neutrophil-dependent hepatocellular necrosis at 7 h (area
of necrosis, 45 ± 3%). Delayed treatment with the caspase 3-like
protease inhibitor Z-Val-Ala-Asp-CH2F (Z-VAD)
(10 mg/kg at 3 h) attenuated apoptosis by 81 to 88% and prevented
liver cell necrosis (
5%). Z-VAD had no effect on the initial
inflammatory response, including the sequestration of neutrophils in
sinusoids. However, Z-VAD prevented neutrophil transmigration and
necrosis. Our data indicate that activation of the caspase 3 subfamily
of cysteine proteases is critical for the development of parenchymal
cell apoptosis. In addition, excessive hepatocellular apoptosis can be
an important signal for transmigration of primed neutrophils
sequestered in sinusoids.
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