Activation of caspase 3 (CPP32)-like proteases is essential for TNF-alpha-induced hepatic parenchymal cell apoptosis and neutrophil-mediated necrosis in a murine endotoxin shock model.
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 microg/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-1beta-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-alpha but not IL-1alphabeta 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 (< or = 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.
Jaeschke
H
H
Department of Pharmacology, Pharmacia & Upjohn, Inc., Kalamazoo, MI 49007, USA. hartmut.w.jaeschke@am.pnu.com
Fisher
M A
MA
Lawson
J A
JA
Simmons
C A
CA
Farhood
A
A
Jones
D A
DA
D000818:Animals; D017209:Apoptosis / Q000276:immunology*; D001427:Bacterial Toxins / Q000008:administration & dosage; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004195:Disease Models, Animal; D004731:Endotoxins / Q000008:administration & dosage; D004789:Enzyme Activation / Q000276:immunology; D005688:Galactosamine / Q000008:administration & dosage; D008099:Liver / Q000201:enzymology / Q000276:immunology / Q000473:pathology*; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D009336:Necrosis; D009504:Neutrophils / Q000276:immunology*; D012475:Salmonella; D012772:Shock, Septic / Q000201:enzymology / Q000276:immunology / Q000473:pathology*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
D001427:Bacterial Toxins; D004731:Endotoxins; D014409:Tumor Necrosis Factor-alpha; C027952:salmonella toxin; D005688:Galactosamine; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases