Enhanced biliary excretion of canalicular membrane enzymes in estrogen-induced and obstructive cholestasis, and effects of different bile acids in the isolated perfused rat liver.
BACKGROUNDS/AIMS: Canalicular membrane enzymes are normally released into bile by partially known processes. This study was undertaken to investigate whether hepatocellular cholestatis induced in rats by ethynylestradiol or obstructive cholestasis produced by complete biliary obstruction for 24 h is associated with an increased release of alkaline phosphatase and gamma-glutamyl transpeptidase into bile, and to clarify how this process is affected by different bile acids. METHODS: The studies were performed in the isolated perfused liver during infusion of sodium taurocholate, taurochenodeoxycholate and tauroursodeoxycholate at increasing rates. RESULTS: Maximum sodium taurocholate, taurochenodeoxycholate and tauroursodeoxycholate secretory rates were decreased in both cholestatic groups (complete biliary obstruction > ethynylestradiol) compared with controls. Maximum biliary outputs of alkaline phosphatase and gamma-glutamyl transpeptidase were significantly increased in the ethynylestradiol group during infusion of sodium taurocholate and taurochenodeoxycholate, but not of tauroursodeoxycholate, and were increased in the complete biliary obstruction group during the infusion of sodium taurocholate and tauroursodeoxycholate but not of taurochenodeoxycholate. The biliary outputs of alkaline phosphatase and gamma-glutamyl transpeptidase showed a significant and direct linear relationship with sodium taurocholate and taurochenodeoxycholate secretory rates in both cholestatic groups. However, only in the complete biliary obstruction group did alkaline phosphatase and gamma-glutamyl transpeptidase excretion show a significant correlation with tauroursodeoxycholate secretory rates. The slope of the line, which indicated the mU of enzyme activity secreted per nmol of sodium taurocholate or taurochenodeoxycholate, was greater for gamma-glutamyl transpeptidase and alkaline phosphatase in both cholestatic groups (ethynylestradiol > complete biliary obstruction) than in the control group. Alkaline phosphatase activity in purified isolated canalicular and sinusoidal membranes was significantly increased in both cholestatic groups (complete biliary obstruction > ethynylestradiol), while gamma-glutamyl transpeptidase activity was unchanged compared with controls. CONCLUSION: The marked increase in sodium taurocholate and taurochenodeoxycholate-mediated release of alkaline phosphatase and gamma-glutamyl transpeptidase into bile in cholestatic rats suggests an increased lability of these intrinsic membrane proteins to the detergent effects of secreted bile acids. It remains to be elucidated whether this phenomenon, which was particularly intense in ethynylestradiol induced cholestasis, is important in the pathogenesis and perpetuation of bile secretory failure. In contrast, tauroursodeoxycholate administration did not result in enhanced biliary excretion of these membrane enzymes, in either the control group or the ethynylestradiol group, supporting the concept that this bile salt lacks the membrane toxicity of common bile acids.