Gastrin inhibits secretin-induced ductal secretion by interaction with specific receptors on rat cholangiocytes.

We assessed the effect of gastrin on ductal secretion in normal and bile duct-ligated (BDL) rats. The effect of gastrin on ductal secretion was examined in the presence of proglumide, a specific antagonist for gastrin receptor (GR). We isolated pure cholangiocytes from normal and BDL rats and assessed gastrin effects on secretin receptor (SR) gene expression and intracellular adenosine 3',5'-cyclic monophosphate (cAMP) levels. We examined the presence of GR mRNA in cholangiocytes by reverse transcription polymerase chain reaction (RT-PCR). In normal or BDL rats, gastrin produced no changes in spontaneous bile secretion. Simultaneous infusion of gastrin inhibited secretin-induced choleresis and bicarbonate output in BDL rats. In the presence of proglumide gastrin did not inhibit secretin-induced choleresis in BDL rats. Gastrin decreased in cholangiocytes from BDL rats 1) SR gene expression and 2) secretin-induced cAMP levels. With the use of RT-PCR, GR mRNA was detected in cholangiocytes. Similar to what is shown for secretin and somatostatin, we propose that the opposing effects of secretin and gastrin on cholangiocyte secretory activity regulate ductal secretion in rats.

Functional expression of the apical Na+-dependent bile acid transporter in large but not small rat cholangiocytes.

BACKGROUND & AIMS: Bile acids interact with cholangiocytes, resulting in cholangiocyte proliferation and increases in ductal bile secretion in large but not small cholangiocytes. It was proposed that for bile acids to exert these effects on cholangiocytes, a specific uptake mechanism must be present in cholangiocytes. The aim of this study was to show the expression of a bile acid transporter in cholangiocytes. METHODS: Small and large cholangiocytes or intrahepatic bile duct units (IBDUs) were isolated from normal rats, and gene expression for the apical Na+-dependent bile acid transporter (ABAT) and the 14-kilodalton ileal cytosolic binding protein (IBABP) was assessed by ribonuclease-protection assays. Tissue and subcellular distribution of bile acid transporters was also studied. [14C]-Taurocholate uptake into cholangiocytes was determined. RESULTS: Both ABAT and IBABP messenger RNAs were detected in large but not small cholangiocytes. By immunohistochemistry, ABAT was present in large but not small cholangiocytes. Immunofluorescence showed ABAT to be present in the apical membrane of large IBDUs. A Na+-dependent saturable uptake of taurocholate was present in large but not small cholangiocytes. CONCLUSIONS: These proteins may mediate bile acid uptake from the duct lumen in large ducts, resulting in modification of canalicular bile secretion and modulation of ductal bile secretion and growth.

gamma-Interferon inhibits secretin-induced choleresis and cholangiocyte proliferation in a murine model of cirrhosis.

BACKGROUND/AIMS: Cholangiocyte proliferation is associated with increased secretin receptor gene expression and secretin-induced choleresis. Since gamma-interferon has antiproliferative effects, we tested the hypothesis that gamma-interferon inhibits ductal proliferation and secretin-stimulated choleresis associated with cirrhosis. METHODS: Mice were treated with 0.1 ml of 25% carbon tetrachloride intraperitoneally twice weekly and 5% alcohol in drinking water for 12 weeks to induce cirrhosis and subsequently gamma-interferon 10(5) intramuscularly was administered daily for 10 weeks. We measured the effects of carbon tetrachloride and gamma-interferon on liver collagen content by morphometric analysis and hydroxyproline content. We measured the effects of gamma-interferon on ductal mass by morphometry and on ductal secretion by assessment of secretin receptor gene expression and secretin-induced choleresis. RESULTS: Compared to controls, there was an increase in liver hydroxyproline content of carbon tetrachloride-treated mice with histologic evidence of cirrhosis. Gamma-interferon treatment significantly decreased collagen liver content with loss of histologic features of cirrhosis. Morphometry revealed an increased number of bile ducts in cirrhotic mice as compared to controls or cirrhotics who received gamma-interferon. Secretin receptor mRNA levels were higher in cirrhotic mice compared to controls but this increase was inhibited by gamma-interferon. Secretin stimulated ductal secretion in cirrhotic mice but not control or cirrhotic mice who received gamma-interferon. CONCLUSIONS: We have established a murine model for cirrhosis and have shown, consistent with our hypothesis, that gamma-interferon decreases collagen content, ductal mass and secretin-induced choleresis incirrhotic mice.

Regrowth of the rat biliary tree after 70% partial hepatectomy is coupled to increased secretin-induced ductal secretion.

BACKGROUND & AIMS: After partial hepatectomy, liver regeneration occurs with the return of hepatocyte mass to normal, Limited data exist regarding the renewal of the biliary tree after partial hepatectomy. This study tested the hypothesis that, after partial hepatectomy, the biliary tree regenerates by proliferation of the remaining cholangiocytes, leading to an increase in secretin-induced ductal bile secretion. METHODS: After 70% partial hepatectomy, cholangiocyte proliferation was assessed in situ by morphometric analysis and In vitro by measurement of 3H-thymidine incorporation. Ductal secretion was estimated by measurement of secretin receptor gene expression and adenosine 3',5'-cyclic monophosphate (cAMP) levels in vitro and by the effect of secretin on ductal bile secretion in vivo. RESULTS: DNA synthesis was undetectable in control cholangiocytes, increased and peaked at day 3 after partial hepatectomy, and returned to normal by day 28. Morphometric analysis showed regrowth of the biliary tree beginning at day 1 with restoration by day 10. The expression of secretin receptor gene and secretin-induced cAMP levels and secretin-induced bicarbonate-rich choleresis increased during the period of bile duct renewal. CONCLUSIONS: After partial hepatectomy, the increase in secretin-induced ductal bile secretion observed during bile duct renewal results from proliferation of remaining cholangiocytes.

Bile acid-dependent vesicular transport of lysosomal enzymes into bile in the rat.

BACKGROUND: Bile acids may stimulate the movement of hepatocyte vesicles and enhance their fusion with the biliary canaliculus. The present study examined the effects of various bile acids on the exocytosis of the contents of hepatocyte lysosomes into the biliary canaliculus. METHODS: The effects of various bile acids on hepatocyte lysosome movement and on exocytosis of the contents of hepatocyte lysosomes into the biliary canaliculus were determined from the distribution of fluorescein isothiocyanate-dextran--labeled lysosomes in hepatocyte couplets and by quantitating biliary lysosomal enzyme output in rats. RESULTS: Hydrophobic as well as hydrophilic and nonmicellar bile acids were found to stimulate to a similar degree the output of lysosomal enzymes into bile, indicating that bile acid-induced change of canalicular or lysosomal membrane fluidity is not responsible for enhanced exocytosis. The taurocholate-dependent increase in lysosomal enzyme excretion was completely blocked by either microtubule or microfilament inhibition, suggesting that these subcellular structures are involved in bile acid-dependent vesicular transport. Fluorescent microscopy studies showed that taurocholate causes a microtubule-dependent translocation of lysosomes towards the canaliculus in hepatocyte couplets, which occurred at the same time as increased output of lysosomal enzymes into bile. CONCLUSIONS: The results suggest that bile acids modulate vesicle traffic towards the canaliculus by a mechanism unrelated to bile acid interaction with the vesicle membrane.

Demonstration of vesicular-dependent bile flow in the sucrose-loaded rat.

The contribution of the hepatocyte vacuolar apparatus to bile fluid formation was assessed by studying the transcellular transport and biliary excretion of the fluid-phase marker sucrose. In rats sucrose-loaded by IP administration of sucrose, electron microscopy showed expansion of the vacuolar apparatus and numerous large lysosomelike structures in hepatocytes. Subcellular distribution studies showed that sucrose was sequestered in lysosomes. Compared with controls, sucrose-loaded rats had a 30% higher (P less than 0.01) bile flow with no change in biliary bile acid or electrolyte concentrations. Administration of colchicine ablated the sucrose-induced choleresis and resulted in parallel changes in biliary secretions of sucrose and lysosomal enzymes. Our data suggest that in the sucrose-loaded rat, the hepatocyte vacuolar apparatus may contribute significantly to bile formation by microtubule-dependent release of fluid into bile by exocytosis.