2001Zhonghua chuanranbing zazhiRequires access

Expression of caspase-3 during hepatocyte apoptosis in the mice with TNF-α induced fulminant hepatic failure

Chaofu Wang

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Abstract

Objective To study the expression of protein and mRNA of caspase-3 during hepatocyte apoptosis in the mice with fulminant hepatic failure (FHF). Methods Liver damage was induced by tumor necrosis factor-α (TNF-α) in D-galactosamine (GalN) sensitized BALB/c mice. Hepatocyte apoptosis was examined by electric microscopy, in site end labeling (ISEL) and DNA electrophoresis on 1.5% agarose gel. The expression of protein and mRNA of caspase-3 was detected by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Results After 1.5 h, 3.5 h, 6 h, and 9 h of administration of GalN and TNF-α, rates of hepatocyte apoptosis were found to be 0.0%, 0.3%, 3.15% and 3.19% respectively. The rates of caspase-3 positive hepatocytes were 0.7%, 4.6% , 17.8% and 24.2% respectively. The relative expressive density of caspase-3 mRNA were 0.26%, 0.29%, 0.41% and 0.38% respectively. Conclusions The expression of caspase-3 is up regulated in the TNF-α induced FHF model and may trigger hepatocyte apoptosis and finally lead to FHF.

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Objective To study the expression of protein and mRNA of caspase-3 during hepatocyte apoptosis in the mice with fulminant hepatic failure (FHF). Methods Liver damage was induced by tumor necrosis factor-α (TNF-α) in D-galactosamine (GalN) sensitized BALB/c mice. Hepatocyte apoptosis was examined by electric microscopy, in site end labeling (ISEL) and DNA electrophoresis on 1.5% agarose gel. The expression of protein and mRNA of caspase-3 was detected by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Results After 1.5 h, 3.5 h, 6 h, and 9 h of administration of GalN and TNF-α, rates of hepatocyte apoptosis were found to be 0.0%, 0.3%, 3.15% and 3.19% respectively. The rates of caspase-3 positive hepatocytes were 0.7%, 4.6% , 17.8% and 24.2% respectively. The relative expressive density of caspase-3 mRNA were 0.26%, 0.29%, 0.41% and 0.38% respectively. Conclusions The expression of caspase-3 is up regulated in the TNF-α induced FHF model and may trigger hepatocyte apoptosis and finally lead to FHF.

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Available abstract

Objective To study the expression of protein and mRNA of caspase-3 during hepatocyte apoptosis in the mice with fulminant hepatic failure (FHF). Methods Liver damage was induced by tumor necrosis factor-α (TNF-α) in D-galactosamine (GalN) sensitized BALB/c mice. Hepatocyte apoptosis was examined by electric microscopy, in site end labeling (ISEL) and DNA electrophoresis on 1.5% agarose gel. The expression of protein and mRNA of caspase-3 was detected by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Results After 1.5 h, 3.5 h, 6 h, and 9 h of administration of GalN and TNF-α, rates of hepatocyte apoptosis were found to be 0.0%, 0.3%, 3.15% and 3.19% respectively. The rates of caspase-3 positive hepatocytes were 0.7%, 4.6% , 17.8% and 24.2% respectively. The relative expressive density of caspase-3 mRNA were 0.26%, 0.29%, 0.41% and 0.38% respectively. Conclusions The expression of caspase-3 is up regulated in the TNF-α induced FHF model and may trigger hepatocyte apoptosis and finally lead to FHF.

Key concepts: Apoptosis, Hepatocyte, Agarose gel electrophoresis, Molecular biology, Fulminant hepatic failure, Messenger RNA, Tumor necrosis factor alpha, Caspase 3

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