Coagulative and Fibrinolytic Activities in Primary Cultured Rat Hepatocytes Treated with Carbon Tetrachloride
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Hepatocyte culture is useful for study of the secretory processes of plasma proteins such as coagulative and fibrinolytic factors, because most coagulative and fibrinolytic factors are synthesized by parenchymal cells in the liver. To investigate<I>in vitro</I>correlation between hepatotoxicity and coagulative and fibrinolytic activities, effects of carbon tetrachloride (CCl<SUB>4</SUB>) on primary cultured rat hepatocytes were studied. Hepatocytes (1 × 10<SUP>6</SUP> cells/ml) were obtained by collagenase perfusion from male Sprague-Dawley 6 week-old rats and suspended in William's E medium with fetal bovine serum. Treatment with CCl<SUB>4</SUB> increased leakage of enzyme activity dose dependently. Urea-nitrogen content and amylase activity decreased significantly with high-dose CCl<SUB>4</SUB> exposure. Coagulative activity in the medium, as expressed by the protein content of fibrin formed by incubation with Ca<SUP>++</SUP> and tranexamic acid (t-AMCHA), increased in 24 and 48 h incubation with 10 and 15 mM CCl<SUP>4</SUP>. Fibrinolytic activity, as the insoluble residual fibrin after incubation without t-AMCHA, also increased. Plasminogen (PLG) content in hepatocytes decreased, and PLG content released into the medium increased in 24 and 48 h incubation with the high-dose CCl<SUB>4</SUB>. These results indicate that the hepatocytes that survive after CCl<SUB>4</SUB> treatment retain coagulative and fibrinolytic activity, although the membrane integrity and capacity for synthesis of urea-nitrogen and amylase are severely limited.
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