Effect of S-adenosyl-L-methionine on ethynylestradiol-induced impairment of bile flow in female rats

Summary Prevention by S-adenosyl-L-methionine (SAMe) of the bile flow reduction induced by ethynylestradiol (EE) is demonstrated by comparing the flow rate and the bile salt concentration of bile in EE-treated animals with that in animals given both EE and SAMe.SAMe was supplied by Bio Research Co., 20060 Liscate (Milano), Italy.     

Influence of phenylbutazone on bile flow in the rat

Summary Phenylbutazone, a well-known enzyme inducer, at a dose of 80 mg·kg^–1 once daily for 8 days increases liver weight and bile flow expressed per g of liver (p<0.01). The bile salt secretory rate is not increased.     

Wirkungen von Gallensäure und Vitamin D auf die Kalziumresorption bei der Ratte

Summary Comparison of the effects of vitamin D and the bile salt Taurochenodeoxycholate on calcium absorption in young rats suggests that bile salt enhancement of calcium absorption occurs mainly in the ileum in normal animals but occurs in both duodenum and ileum in rachitic rats. Also suggested is that bile salts mainly assist the entry of calcium into mucosal cells from the gut lumen while vitamin D chiefly aids the egress of calcium from the cells into the extracellular fluid.     

Choleretic and cholestatic effects of infused bile salts in the rat

Summary In rats, at low infusion rates taurocholate (TC), taurochenodeoxycholate (TCDC) and taurodeoxycholate (TCD) each produced an increase in bile flow of 20–50%. However, at high infusion rates (5–20 moles min^–1 kg^–1) the cholestatic effects of the bile salts were revealed and the relative toxicity of the bile salts was seen to be TDC>TCDC>TC.     

Expansion of the bile acid pool changes the biliary transport characteristics of centrizonal hepatocytes

We investigated whether acinar differences in taurocholate transport are responsible for the increased maximal secretory rate observed after expansion of the bile acid pool. The bile acid pool was expanded by cholate feeding for four days. Periportal and centrizonal hepatocytes were then probed by ante- and retrograde liver perfusion, respectively. In control animals, secretory rate constant (alpha 1) averaged 0.439 +/- 0.123 and 0.104 +/- 0.035 min-1 during ante- and retrograde perfusion, respectively, in the absence of exogenous taurocholate. These values did not significantly change when taurocholate was infused. In cholate-fed animals, alpha 1 was comparable during antegrade perfusion but was significantly reduced (0.038 +/- 0.035, p less than 0.05) during retrograde perfusion in the absence of exogenous taurocholate, presumably owing to induction of cytosolic bile acid binding proteins. During loading with exogenous taurocholate, by contrast, alpha 1 was significantly accelerated (0.252 +/- 0.026; p less than 0.01) in centrizonal hepatocytes from bile-acid fed rats. Expansion of the bile acid pool is able to change the bile salt secretory characteristics of centrizonal hepatocytes toward those of periportal ones.     

Bile Acids: Chemistry, Pathochemistry, Biology, Pathobiology, and Therapeutics

Bile acids and bile alcohols in the form of their conjugates are amphipathic end products of cholesterol metabolism with multiple physiological functions. The great variety of bile acids and bile alcohols that are present in vertebrates are tabulated. Bile salts have an enterohepatic circulation resulting from efficient vectorial transport of bile salts through the hepatocyte and the ileal enterocyte; such transport leads to the accumulation of a pool of bile salts that cycles between the liver and intestine. Bile salt anions promote lipid absorption, enhance tryptic cleavage of dietary proteins, and have antimicrobial effects. Bile salts are signaling molecules, activating nuclear receptors in the hepatocyte and ileal enterocyte, as well as an increasing number of G-protein coupled receptors. Bile acids are used therapeutically to correct deficiency states, to decrease the cholesterol saturation of bile, or to decrease the cytotoxicity of retained bile acids in cholestatic liver disease.     

Choleretic and cholestatic effects of infused bile salts in the rat.

In rats, at low infusion rates taurocholate (TC), taurochenodeoxycholate (TCDC) and taurodeoxycholate (TCD) each produced an increase in bile flow of 20-50%. However, at high infusion rates (5-20 mumoles min-1kg-1) the cholestatic effects of the bile salts were revealed and the relative toxicity of the bile salts was seen to be TDC greater than TCDC greater than TC.     

The inability of nuclear dehydrogenating clostridia to oxidize bile salt hydroxyl groups.

In a survey of the intracellular bile salt oxidoreductase activity in fecal bacteria, 16 strains of nuclear dehydrogenating clostridia and 2 strains of non-nuclear dehydrogenating C. paraputrificum were demonstrated unable to oxidize cholate at any of the 3 OH groups. Since nuclear dehydrogenation at the delta-1 and delta-4 position requires a 3-oxo precursor steroid, it appears that these organisms require the presence of a 3 alpha-hydroxysteroid dehydrogenating organism for nuclear dehydrogenation.     

Solubilization of unconjugated bilirubin by bile salts.

Freshly precipitated unconjugated bilirubin (UCB) is solubilized rapidly and to a large extent by the sodium salts of di- and trihydroxy bile acids. The solubilization effect depending on bile salt concentration, pH and ionic strength is based on micellar mechanisms.     

Solubilization of unconjugated bilirubin by bile salts

Summary Freshly precipitated uncojugated bilirubin (UCB) is solubilized rapidly and to a large extent by the sodium salts of di- and trihydroxy bile acids. The solubilization effect depending on bile salt concentration, pH and ionic strength is based on micellar mechanisms.     

The inability of nuclear dehydrogenating clostridia to oxidize bile salt hydroxyl groups

Summary In a survey of the intracellular bile salt oxidoreductase activity in fecal bacteria, 16 strains of nuclear dehydrogenating clostridia and 2 strains of non-nuclear dehydrogenatingC. paraputrificum were demonstrated unable to oxidize cholate at any of the 3 OH groups. Since nuclear dehydrogenation at the -1 and -4 position requires a 3-oxo precursor steroid, it appears that these organisms require the presence of a 3-hydroxysteroid dehydrogenating organism for nuclear dehydrogenation.This work is supported by the National Cancer Institute of Canada (IAM) and by the Cancer Research Campaign (MJH).     

Intraluminal bile salt increases rate of firing in afferent fibers from the small intestine of the rat

Perfusion of a rat intestinal segment with a solution containing sodium deoxycholate (8 mM) increases the rate of firing in periarterial afferent nerves from the gut. This observation indirectly supports our earlier proposal that bile salt evokes a net fluid secretion in the small intestine via an activation of the enteric nervous system.     

Gallen und Gallensäuren Papierchromatographische Untersuchungen

Summary In cases of bile-scretion disturbances substitution-therapy can be performed using suitable animal biles equivalent to human bile. Comparative investigations have been made to establish which animal bile is in practice the most suitable for bile substitutiontherapy based on its bile acid components.A new paper partition chromatographic method has been applied for separation and identification of free bile acids. Ascending development was used on Schleicher & Schüll 2043b Mgl paper impregnated with 20 v/v% propylene glycol in chloroform. The xylene-methylethylketone 1:1 solvent system gave good separation. The bile acids can be detected by immersing the chromatograms in 20 w/v% SbCl₃ in chloroform followed by drying and heating for 5–10 min at 100–110°C. The spots show intense reddish-violet or blue fluorescence in filtered UV-light (see Table 1).These investigations have shown that human and ox biles are, in contrast to pig bile, physiologically related biological substances regarding their bile acid components.     

Circadian rhythm of bile secretion in the rat.

In rats the bile flow and the estimated bile acid independant flow (BAIF) were significantly lower at 17.00 h than at 08.00 and 24.00 h. The decrease in BAIF paralleled the decrease in liver weight. Bile acid excretion was not different.     

Dietary vitamin E does not protect from endotoxin-induced hepatic microvascular dysfunction

Starting from the concept that lipopolysaccharide (LPS)-associated hepatotoxicity involves the action of reactive oxygen species, the present study was conducted to test whether vitamin E, a lipophilic antioxidant, prevents LPS-induced hepatic microvascular dysfunction and liver injury. Fifty-two rats were divided into three groups and fed diets containing 0 (n=16), 75 (n=18) or 8000 mg (n=18) α-tocopherol acetate/kg food for four weeks. At 1 h and 6 h after intravenous LPS-exposure (10 mg/kg E. coli LPS) hepatic microvascular response and liver injury were assessed by the analysis of Kupffer cell phagocytic activity, leukocyte-endothelial cell interaction and nutritive sinusoidal perfusion (intravital fluorescence epi- illumination technique) as well as bile flow, serum liver enzyme activities and tissue histomorphology. In animals fed with 75 mg vitamin E/kg (standard diet), LPS caused hepatic Kupffer cell activation (increased phagocytic activity) and hepatic microvascular leukocyte activation, with stasis in sinusoids and adherence in postsinusoidal venules (1 h) followed by leukocytic infiltration into tissue (6 h) and progredient sinusoidal perfusion failure (6 h). Hepatic microvascular injury was accompanied by reduced bile flow and enhanced liver enzyme release. Vitamin E-enriched diet (8000 mg/kg) and even vitamin E-deficient diet did not significantly affect LPS-induced hepatic microvascular cell activation and perfusion failure. Thus, we conclude, that vitamin E is not effective to protect from endotoxin-induced hepatic microvascular dysfunction. Received 7 November 1996; received after revision 30 December 1996; accepted 20 January 1997     

Influence of common bile duct cannula size on maximal secretory rate of taurocholate in the rat

Summary The common bile duct of male Sprague-Dawley rats was cannulated with either PE 10 or PE 50 tubing. Maximal secretory rate of taurocholate averaged 389±67 (SD) and 657±115 nmoles·min^–1·g liver^–1 in the PE 10 and PE 50 group, respectively (p<0.005). Maximal bile secretory pressure was significantly higher in the PE 10 group (240±28 vs 174±8 mm H20; p<0.005). When the maximal secretory rate was exceeded, bile flow decreased in both groups but this was accompanied with a decrease in maximal bile secretory pressure in the PE 10 group only. Maximal secretory rate of bile salts is markedly influenced by experimental technique. Use of small caliber common bile duct cannulae leads to partial obstruction and decreases the apparent maximal secretory rate for taurocholate.Acknowledgments. J. Reichen was the recipient of a Faculty Development Award in Clinical Pharmacology from The Pharmaceutical Manufacturer's Association Foundation, and is the recipient of a Research Career Development Award (KO4 AM 1189) from the National Institutes of Health. Supported by National Institutes of Health grant RO1 AM 27597.     

Effect of food and light schedule on bile flow in the rat

In the rat after food intake, whether during the dark or light period, bile flow increases. Food intake seems to be a major factor in the circadian rhythm of bile secretion.     

Effect of food and light schedule on bile flow in the rat

Summary In the rat after food intake, whether during the dark or light period, bile flow increases. Food intake seems to be a major factor in the circadian rhythm of bile secretion.     

ABCB4 missense mutations D243A,K435T,G535D,I490T,R545C,and S978P significantly impair the lipid floppase and likely predispose to secondary pathologies in the human population

Bile salts are natural detergents required to solubilise dietary fat and lipid soluble vitamins. They are synthesised in hepatocytes and secreted into the luminal space of the biliary tree by the bile salt export pump (BSEP), an ATP-binding cassette (ABC) transporter in the canalicular membrane. BSEP deficiency causes cytotoxic accumulation of bile salts in the hepatocyte that results in mild-to-severe forms of cholestasis. The resulting inflammation can also progress to hepatocellular cancer via a novel mechanism involving upregulation of proliferative signalling pathways. A second ABC transporter of the canalicular membrane is also critical for bile formation. ABCB4 flops phosphatidylcholine into the outer leaflet of the membrane to be extracted by bile salts in the canalicular space. These mixed micelles reduce the detergent action of the bile salts and protect the biliary tree from their cytotoxic activity. ABCB4 deficiency also causes cholestasis, and might be expected to cause cholangitis and predispose to liver cancer. Non-synonymous SNPs in ABCB4 have now been described in patients with liver cancer or with inflammatory liver diseases that are known to predispose to cancer, but data showing that the SNPs are sufficiently deleterious to be an etiological factor are lacking. Here, we report the first characterisation at the protein level of six ABCB4 variants (D243A, K435T, G535D, I490T, R545C, and S978P) previously found in patients with inflammatory liver diseases or liver cancer. All significantly impair the transporter with a range of phenotypes exhibited, including low abundance, intracellular retention, and reduced floppase activity, suggesting that ABCB4 deficiency is the root cause of the pathology in these cases.     

Intraluminal bile salt increases rate of firing in afferent fibers from the small intestine of the rat

Summary Perfusion of a rat intestinal segment with a solution containing sodium deoxycholate (8 mM) increases the rate of firing in periarterial afferent nerves from the gut. This observation indirectly supports our earlier proposal that bile salt evokes a net fluid secretion in the small intestine via an activation of the enteric nervous system.This study was supported by grants from the Swedish Medical Research Council (2855), The Medical Council of Swedish Life Insurance Companies and the Faculty of Medicine, University of Göteborg.