Role of ductular bile water reabsorption in canine bile secretion

To quantitate ductular and/or ductal bile water reabsorption and to define its role in hepatic bile formation, spontaneous and taurocholate-stimulated bile flow and composition were measured in anesthetized dogs postprandially (n = 4) and after a 24-hour (n = 4) and a 48-hour (n = 4) fasting period. Spontaneous bile flow in fed dogs (0.344 +/- 0.116 microliter/min/gm liver tissue) was significantly higher than that in 24-hour (0.179 +/- 0.046 microliter/min/gm) and 48-hour fasted animals (0.096 +/- 0.085 microliter/min/gm). One 48-hour fasted dog was virtually cholestatic (spontaneous bile flow 0.004 microliter/min/gm), and another 48-hour fasted animal had a mean bile flow rate as low as 0.036 microliter/min/gm. Calculated ductular bile water reabsorption (carbon 14-erythritol biliary clearance minus bile flow) was the same in the three groups of animals (0.166 +/- 0.056, 0.208 +/- 0.033, and 0.204 +/- 0.055 microliter/min/gm, respectively). Administration of sodium taurocholate (0.5 to 2.0 mumol/min/kg) revealed intact canalicular secretory capacity in all dogs, and no change in bile water reabsorption rate was observed during choleresis. Bile chloride and bicarbonate concentrations during spontaneous secretion were significantly lower in 48-hour fasted dogs when compared with fed animals. In the two 48-hour fasted dogs with a very low rate of spontaneous bile flow, chloride and bicarbonate concentrations in hepatic bile were as low as those seen in gallbladder bile (5 to 10 mEq/L) and increased during taurocholate choleresis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Biliary excretion and choleretic effect of cefmetazole in rats.

The effect of cefmetazole, a broad-spectrum cephalosporin, on bile flow and composition in rats was studied. Intravenous injection of cefmetazole at doses ranging from 40 to 400 mumol/kg of body weight led to an increase in its biliary concentration and excretion rate, with a maximum at 30 min after injection. Excretion of cefmetazole into bile was associated with a marked choleresis. The magnitude of the increase in bile flow was dose dependent, with a maximal increase at a dose of 200 mumol/kg. Cefmetazole administration did not affect the secretion of bile acids or their osmotic activities, whereas the bile acid-independent bile flow increased by 49% at a dose of 200 mumol/kg. Cefmetazole administration at a dose of 200 mumol/kg significantly increased the biliary outputs of sodium, potassium, chloride, and bicarbonate (+36, +56, +28, and +31%, respectively) compared with outputs of controls. A linear relationship was observed between bile flow and cefmetazole excretion, 44 microliters of bile being produced per mumol of cefmetazole excreted into bile. Our results demonstrate that cefmetazole induces choleresis by stimulating bile acid-independent bile flow. This effect appears to be partly due to the osmotic properties of cefmetazole transported into bile.     

Prolactin regulates maternal bile secretory function post partum.

The single-pass isolated perfused rat liver was used to assess bile secretory function in pregnancy and the post partum period and to evaluate the regulatory role of prolactin. Nonpregnant controls, post partum rats (48 hr post partum), pregnant rats (19-20 days of pregnancy) and pregnant rats treated with bromocriptine (3 mg/kg/day, s.c.) from day 20 of pregnancy until 48 hr post partum to block prolactin secretion were examined. In a separate set of experiments, ovariectomized rats were implanted with osmotic minipumps containing solvent alone or solvent plus varying concentrations of ovine prolactin (oPRL). Livers were isolated, [3H]taurocholate (TC) was infused at increasing concentrations and bile flow, bile acid secretory rate and hepatic clearance of TC from the perfusate were determined. Maximal bile flow (microliters/min/g liver) in response to infusion of TC was significantly higher in post partum rats (3.4 +/- 0.3) than in pregnant rats (1.4 +/- 0.3) and bromocriptine-treated post partum rats (1.7 +/- 0.3). The maximal bile acid secretory rate (SRm, nmol/min/g liver) in post partum rats was 276 +/- 19 and was significantly greater than SRm in pregnant (103 +/- 26) and bromocriptine-treated post partum rats (142 +/- 25). Hepatic clearance (ml/min) was significantly greater in post partum rats than in pregnant, control and bromocriptine-treated post partum rats at the highest TC concentration. Infusion of oPRL at doses of 100, 250 and 500 micrograms/day significantly increased maximal bile flow and SRm in a dose-dependent manner; these measures were increased to 4.38 +/- 0.21 microliters/min/g liver and 338.2 +/- 16.3 nmol/min/g liver, respectively, in rats treated with 500 micrograms/day oPRL.(ABSTRACT TRUNCATED AT 250 WORDS)     

Rat model of mild stenosis of the common bile duct

Several techniques for developing incomplete obstruction of the common bile duct have been described but none of them properly represents a compression or constriction of the bile duct. In this study, a mild stenosis of the common bile duct was achieved in the rat by means of a double ligature including a cannula that could be easily slipped out of the ligatures. Sham-operated rats were used as controls. The studies, performed 7–10 days postoperatively, indicated that in ligated rats a duct constriction was produced, made evident by an increase of the biliary pressure, an upstream dilatation of the bile duct, an increase of the liver volume constituted by portal tracts, and ductular proliferation. Serum parameters were practically similar in ligated and control rats, except for a slight increase in serum bilirubin. Following intravenous injection of sodium taurocholate there were rapid increases of bile flow and bile salt output in both groups, but choleresis induced by sodium taurocholate was higher in ligated rats than in controls. The clearances of [¹⁴C]erythritol and [¹⁴C]sucrose suggested that ductular water contributing to bile flow and changes in biliary permeability were not involved in ligated rats. The limited repercussion of humoral effects and hepatic behaviour seen in ligated rats despite the morphological alterations induced make the mild stenosis of the bile duct a good model for the study of early stages of compression or constriction of the biliary tract.     

The effect of sodium taurocholate on the hepatic metabolism of sulfobromophthalein sodium (BSP). The role of bile flow

The influence of bile salts on the hepatic metabolism of sulfobromophthalein sodium (BSP) was studied in the perfused rat liver. During sodium taurocholate infusions, hepatic uptake of BSP from plasma was increased and appeared to be related to an enhanced transit of BSP from liver into bile. BSP-glutathione conjugation was not affected by the bile salt infusions, although bile salts inhibited the enzyme system in vitro.The major effect of bile salts was to increase the BSP transport maximum (Tm). When sodium taurocholate was infused in saline at a rate of 30 mumoles/hr, both bile flow and the BSP Tm increased, and remained at peak levels of 1.5 +/-0.12 mul/min per g liver and 21 +/-3.0 mug/min per g liver, respectively. In contrast, during saline infusion alone both levels were significantly lower (1.06 +/-0.17 mul/min per g liver and 15.8 +/-4.16 mug/min per g liver, respectively), and both fell progressively after the 2nd hr of perfusion. This decline in bile flow and BSP Tm was associated with a decrease in biliary bile salt excretion and was reversed by adding bile salts to the perfusate. Since the biliary concentration of BSP remained within a narrow range in all experiments, the BSP Tm was primarily determined by the rate of bile flow.Dependence of BSP Tm on the rate of bile production was further confirmed by changing the temperature of the perfusate during a constant infusion of taurocholate. BSP Tm paralleled temperature-induced changes in bile flow irrespective of changes in the level of bile salt excretion.Since the biliary concentration of BSP remained within a narrow range in all experiments, the concentrating capacity for BSP in bile may be the major limiting factor in BSP transport. Thus two independent factors appear to determine the BSP Tm: the bile BSP concentration, and the rate of bile production.Because taurocholate enhanced BSP transport only when it increased bile production, its effect on the BSP Tm appears to be attributable to its choleretic properties.     

The effect of complete biliary obstruction on bile flow and bile acid excretion: postcholestatic choleresis in the rat.

Bile secretory function was studied in rats subjected to a 7-day obstructive cholestasis induced by complete common duct obstruction. Bile flow and bile acid excretion were examined during bile depletion, following the release of the biliary obstruction, and during the infusion of sodium taurocholate at submaximal and saturating rates. A highly significant increase, greater than 100%, in bile flow was evident in cholestatic rats at any bile acid excretory rate, when compared to control sham-operated rats. 14C-erythritol clearance measurements performed during bile depletion and during the infusion of taurocholate suggest that bile flow was mainly of canalicular origin in cholestatic rats. Estimated taurocholate transport maximum (mumol/min per rat) was not statistically different between cholestatic and control rats. However, significantly greater taurocholate plasma levels at Tm in cholestatic rats suggest a decreased efficiency of the bile acid transport process. In addition, the relationship between bile flow and bile acid excretion was found to be nonlinear at low bile acid excretory rates in cholestatic rats. Thus important changes in bile formation occurred in rats subjected to temporary obstructive cholestatis, which differ from those observed in other models of cholestasis that are associated to a reduction in bile flow and bile acid transport capacity.     

Abolition of valproate-derived choleresis in the Mrp2 transporter-deficient rat

Valproic acid (VPA) is a major therapeutic agent in the treatment of epilepsy and other neurological disorders. It is metabolized in humans and rats primarily along two pathways: direct glucuronidation to yield the acyl glucuronide (VPA-G) and beta-oxidation. We have shown much earlier in the Sprague-Dawley rat that i.v. administration of sodium valproate (NaVPA) caused a marked choleresis (mean of 3.3 times basal bile flow after doses of 150 mg/kg), ascribed to the passive osmotic flow of bile water following excretion of VPA-G across the canalicular membrane. Active biliary pumping of anionic drug conjugates across the canalicular membrane is now believed to be attributable to transporter proteins, in particular Mrp2, which is deficient in the TR(-) (a mutant Wistar) rat. In the present study, normal Wistar and Mrp2-deficient TR(-) rats were dosed i.v. with NaVPA at 150 mg/kg. In the Wistar rats, there was a peak choleretic effect of about 3.2 times basal bile flow, occurring at about 30 to 45 min postdose (as seen previously with Sprague-Dawley rats). In TR(-) rats given the same i.v. dose, there was no evidence of postdose choleresis. The choleresis was correlated with the excretion of VPA-G into bile. In Wistar rats, 62.8 +/- 7.7% of the NaVPA dose was excreted in bile as VPA-G, whereas in TR(-) rats, only 2.0 +/- 0.6% of the same dose was excreted as VPA-G in bile (with partial compensatory excretion of VPA-G in urine). This study underlines the functional (bile flow) consequences of biliary transport of xenobiotic conjugated metabolites.     

Effects of chlorpromazine hydrochloride on bile salt synthesis, bile formation and biliary lipid secretion in the Rhesus monkey: a model for chlorpromazine-induced cholestasis

We studied the acute effects of intravenous infusions of chlorpromazine hydrochloride on bile salt synthesis, bile formation and biliary lipid secretion in the alert female Rhesus monkey prepared with a total biliary fistula and in a steady bile salt secretory state. In twelve studies (three animals), five doses of radiolabelled chlorpromazine hydrochloride (1-10 mg identical to 2.8-28 mumol/kg) were infused intravenously for 1 h in random order. Cholestasis was induced within minutes in all experiments. The radiolabel appeared rapidly in bile, with similar recoveries in bile and urine and a 90% total cumulative output in 4 days. Both bile flow, bile salt and other biliary lipid outputs were inhibited in a dose related and reversible manner. The apparent bile salt independent bile flow was consistently abolished, and a prompt return to basal values occurred when biliary concentration of the drug and metabolities fell below 1-2 mM. When chlorpromazine hydrochloride was infused at three doses (2.5, 5.0 and 10.0 mg identical to 7-28 mumol/kg) during constant intravenous infusion of 14C sodium taurocholate (300 mumol/h), bile flow, total bile salt output and 14C taurocholate output decreased within minutes. This was accompanied by a progressive rise in the serum 14C taurocholate concentration. After 90 min the taurocholate specific activity in bile increased significantly indicating that bile salt synthesis was inhibited. Secretion of retained bile salts and reversal of inhibition of bile salt synthesis occurred with time: the course of both events was correlated with the dose of the drug. Thus, in monkeys, chlorpromazine hydrochloride induces reversible, dose related cholestasis suppression of the bile salt dependent and independent flow, inhibition of bile salt synthesis and impairment of biliary lipid secretion. We suggest that these effects are due to both bile salt-chlorpromazine interactions and the effect of the latter on canalicular and other membranes.     

The effect of complete biliary obstruction on bile secretion. Studies on the mechanisms of postcholestatic choleresis in the rat

It has been previously shown that rats subjected to obstructive cholestasis demonstrate in the postcholestatic period, after release of common duct obstruction, a marked increase in canalicular bile flow relative to bile acid excretion. To characterize this phenomenon further, we investigated whether changes in canalicular permeability and in the activity of (Na+-K+)-ATPase in isolated liver surface membranes are associated with postcholestatic choleresis. With this purpose, the clearances of 14C-erythritol and 3H-insulin were simultaneously measured in rats subjected to a 3-day obstructive cholestasis and in controls, during spontaneous choleresis as well as during the intravenous infusion of sodium taurocholate at both submaximal and saturating rates. In additional groups of bile duct-ligated rats and controls, liver surface membrane fractions were isolated, and the activity of appropriate marker enzymes and (Na+-K+)-ATPase was determined. In both groups 14C-erythritol clearance closely approximated total bile flow, suggesting that bile flow was of canalicular origin. However, cholestatic rats showed a sixfold increase in 3H-inulin clearance compared to controls. These results suggest that canalicular permeability to inulin is markedly increased in cholestatic rats. On the other hand, (Na+-K+)-ATPase activity was significantly higher in cholestatic rats than in controls in both homogenate (1.27 +/- 0.07 and 0.89 +/- 0.07 U/mg of protein, respectively, p less than 0.001) and liver surface membranes (22.6 +/- 1.2 and 17.5 +/- 1.2 U/mg of protein, respectively, p less than 0.001). Thus enhanced choleretic response to bile acids in the postcholestatic period is associated with an increased permeability of canalicular structure to inulin and with a significant increase in both homogenate and surface membrane (Na+-K+)-ATPase activity. In addition, this study points out some important differences between bile secretory function of rats subjected to obstructive cholestasis and that described in models of bile secretory failure induced by drugs or monohydroxy-bile acids.     

Effects of troleandomycin administration on cholesterol 7 alpha-hydroxylase activity and bile secretion in rats

Repeated administration of troleandomycin increased bile flow but decreased the biliary secretion of bile acids in rats. The increased bile flow was associated with a parallel increase in the biliary clearance of [14C]erythritol. Analysis of the relationship between bile flow and bile acid secretion indicated that, for any given rate of bile acid secretin, bile flow was higher in troleandomycin-treated rats than in control rats. The increased bile flow was associated with an increased activity of Na+,K+-adenosine triphosphatase in liver plasma membranes. The decreased bile acid secretion into bile was associated with a similar decrease in the bile acid pool size, a decreased bile acid synthesis rate and a decreased activity of microsomal cholesterol 7 alpha-hydroxylase. The concentration of bile acids in serum, the hepatic extraction ratio of [3H]taurocholate and its biliary transport maximum were not modified. It is concluded that repeated administration of troleandomycin increases the canalicular bile acid-independent flow but decreases the activity of cholesterol 7 alpha-hydroxylase, the synthesis, the pool size and the biliary secretion rate of bile acid in rats.     

The effect of thyroid hormone on bile salt-independent bile flow and Na+, K+ -ATPase activity in liver plasma membranes enriched in bile canaliculi.

The relationship between bile salt-independent canalicular flow and ATPase activity in liver plasma membranes (LPM) enriched in bile canaliculi, was studied in control, hyperthyroid, and hypothyroid rats. Canalicular bile production was significantly increased in hyperthyroid rats (3.19 +/- 0.23 mul/min per g liver) compared to controls (2.27 +/- 0.24 mul/min per g liver), while it diminished in hypothyroid animals (1.58 +/- 0.17 mul/min per g liver). Although bile salt excretion was also increased in hyperthyroid animals (62.4 +/- 13.3 vs. 41.2 +/- 8.4 nmol/min per g liver), the stimulation in canalicular secretion was primarily related to enhancement of the bile salt-independent fraction of flow (2.47 mul/min per g liver in hyperthyroid rats vs. 1.67 mul/min per g liver in controls). LPM Na+, K+-ATPase activity doubled in hyperthyroid animals (21.5 +/- 5.8 vs. 10.7 +/- 3.1 mumol Pi/mg protein per h) while Mg++-ATPase activity remained unchanged and 5'-nucleotidase activity increased to a small but significant extent. In hypothyroid rats, bile salt excretion remained unchanged from control values so that the reduced secretion was entirely secondary to an inhibition of bile salt-independent secretion (1.19 mul/min per g liver). Na+, K+-ATPase activity in the LPMs from hypothyroid animals decreased by nearly 50% (5.4 +/- 1.6 mumol Pi/mg protein per h), although comparable reductions in the specific activity of Mg++-ATPase and 5'-nucleotidase were also observed. Administration of L-thyroxine to hypothyroid animals restored both bile salt-independent canalicular secretion and membrane enzymes to control values within 2 and 4 days, respectively. Sodium dodecyl sulfate gel electrophoresis demonstrated no significant changes in LPM protein fractions from any of the treatment groups. These studies indicate that thyroid hormone has a parallel effect on bile salt-independent canalicular secretion and LPM Na+, K+-ATPase activity, supporting the hypothesis that Na+ transport and Na+, K+-ATPase may be determinants of bile salt-independent canalicular flow.     

Bile Acid-Induced Increase in Bile Acid-Independent Flow and Plasma Membrane NaK-ATPase Activity in Rat Liver

Previous studies showed that in rats with obstruction of the bile ducts draining the median and left hepatic lobes, and in rats with normal bile ducts in which the bile acid pool size and secretion were augmented by 48-h intraduodenal infusion of taurocholate, bile acid flux through secreting hepatocytes was increased. Under these conditions, taurocholate transport maximum exhibited a time-dependent adaptation to increased secretory load.Unexpectedly, bile acid-independent canalicular flow in these experimental models also was found to be increased when measured at 48 h. Relative to controls, bile acid-independent flow per gram of nonobstructed liver was increased approximately threefold in selectively obstructed rats and 43% in bile acid-loaded rats with normal ducts. In rats infused with bile acids at similar rates for only 16 h, no increase was observed. Studies with [(14)C]erythritol suggested that the increased bile flow under these conditions was of canalicular origin.NaK-ATPase activity in canaliculi-enriched liver plasma membrane preparations from the nonobstructed lobes of selectively obstructed rats and from 48-h bile acid-loaded rats was increased by 47% and 52%, respectively, relative to controls, but was not increased in membranes from 16-h bile acid-loaded rats. Canalicular membrane 5'-nucleotidase and Mg ATPase also were increased.These studies show that augmented bile acid flux through secreting liver causes an adaptive increase in bile acid-"independent" flow and in the activity of canalicular membrane enzymes. The mechanism by which bile acids modulate this and previously reported aspects of bile secretion remains to be elucidated.     

Reconstruction of intrahepatic bile ducts in congenital biliary atresia.

Macroserial reconstruction of the main intralobular bile ducts was made in 7 cases of biliary atresia; 2 cases of type I, 1 case of type II and 4 cases of type III according to Kasai's classification. From the results of these reconstruction studies, it was confirmed that the main interlobular bile ducts are usually patent through the liver regardless of the type of atresia of the extrahepatic bile ducts. A microserial reconstruction of bile ducts and ductules of a small portal tract performed in one case disclosed that a number of ductules make a network around the main duct and have some communications with the main duct. These results were compatible with the fact that active excretion of bile was obtained in many postoperative patients with biliary atresia. As observed in one case of the present series, postoperative complication of severe ascending cholangitis seemed to be an important cause of destruction or disappearance of intrahepatic bile ducts, which has also seen in older infants with this disease without complicaion of cholangitis. In view of these facts the operation of an early stage of life is recommended in biliary atresia.     

Relationship between bile flow and Na+, K+-adenosinetriphosphatase in liver plasma membranes enriched in bile canaliculi.

The relationship between bile flow and Na+,K+-ATPase activity in liver plasma membranes enriched in bile canaliculi was studied in rats treated with ethinyl estradiol, phenobarbital, or 20-methyl cholanthrene. In comparison with controls (1.49+/-0.12 microliter/min per g liver), bile flow was significantly diminished by ethinyl estradiol, increased by phenobarbital, and unchanged by 20-methyl cholanthrene or the solvent, propanediol (0.92+/-0.31, 2.50+/-0.21, 1.62+/-0.18, and 1.64+/-0.30 microliter/min per g liver, respectively). The corresponding values for canalicular Na+,K+-ATPase activity were 80.7+/-19.2, 50.0+/-18.4, 231.7+/-42.6, 82.7+/-30.7, and 143.6+/-55.3 micronmol Pi/h per g liver. Canalicular Na+,K+-ATPase activity was significantly correlated (r=0.785, n=31) with bile flow. These findings support the hypothesis that a fraction of bile flow is related to Na+,K+-ATPase activity and canalicular Na+ transport.     

Separate transport systems for biliary secretion of sulfated and unsulfated bile acids in the rat.

Biliary secretion of 3 alpha-sulfated bile acids has been studied in Wistar rats with an autosomal recessive defect in the hepatic transport of bilirubin. Liver function, established by measurement of various enzymes in plasma, by enzyme histochemical methods, and by electron microscopy, appeared to be normal in these rats. Serum levels of unconjugated, monoglucuronidated, and diglucuronidated bilirubin were 0.62, 1.62, and 6.16 mumol/liter, respectively, compared with 0.17, 0.08, and 0.02 mumol/liter in control rats. Biliary bilirubin secretion was strongly reduced in the mutant animals: 0.21 +/- 0.03 vs. 0.39 +/- 0.03 nmol/min per 100 g body wt in control rats. Despite normal biliary bile acid output, bile flow was markedly impaired in the mutant animals, due to a 53% reduction of the bile acid-independent fraction of bile flow. The transport maximum for biliary secretion of dibromosulphthalein (DBSP) was also drastically reduced (-53%). Biliary secretion of intravenously administered trace amounts of the 3 alpha-sulfate esters of 14C-labeled taurocholic acid (-14%), taurochenodeoxycholic acid (-39%), taurolithocholic acid (-73%), and glycolithocholic acid (-91%) was impaired in the jaundiced rats compared with controls, in contrast to the biliary secretion of the unsulfated parent compounds. Hepatic uptake of sulfated glycolithocholic acid was not affected in the jaundiced animals. Preadministration of DBSP (15 mumol/100 g body wt) to normal Wistar rats significantly impaired the biliary secretion of sulfated glycolithocholic acid, but did not affect taurocholic acid secretion. We conclude that separate transport systems in the rat liver exist for biliary secretion of sulfated and unsulfated bile acids; the sulfates probably share secretory pathways with the organic anions bilirubin and DBSP. The described genetic defect in hepatic transport function is associated with a reduced capacity to secrete sulfated bile acids into bile; this becomes more pronounced with a decreasing number of hydroxyl groups on the sulfated bile acid's molecule.     

Characterization of SC2644-induced choleresis in the dog. Evidence for canalicular bicarbonate secretion

The biliary clearances of [14C]erythritol (Cery) and [3H]mannitol (Cmann) were measured simultaneously in dogs, first during choleresis induced by varying doses of sodium taurocholate and then by SC2644. Cery increased equally with the increases in bile flow induced by both compounds. Mannitol entry into bile, however, was partially restricted; deltaCmann/deltabile flow averaged 0.66 and 0.68 for taurocholate- and SC2644-induced flows, respectively. These findings suggest a common canalicular site of origin of the increased bile flow. Electrolyte composition was quite different in the increments, however. The bicarbonate concentration in the SC2644-induced increment of bile (65.8 microEq/ml) was three times higher than that associated with bile stimulated by taurocholate. SC2644- and taurocholate-induced biles were virtually isosmotic. These results in concert with other observations suggest a canalicular mechanism for bicarbonate entry into the biliary tree. Stimulation by SC2644 of ductal chloride bicarbonate exchange cannot be excluded, however.     

Biliary excretion of lecithin and cholesterol in the dog

The biliary excretion rates of bile acid, lecithin, and cholesterol were measured in unanesthetized dogs after interruption of enterohepatic circulation and during infusions of sodium taurocholate, sodium glycocholate, sodium dehydrocholate, SC2644 (a bicyclic organic acid with high choleretic potency), and secretin. Both lecithin output and cholesterol output were directly related to bile acid excretion rate. The curves describing these relationships were concave downward. Molar concentration ratios of lecithin-to-bile acid declined gradually from approximately 0.4 to 0.2 as bile acid output increased from approximately 1 to 70 mumoles/min. Cholesterol-to-lecithin molar ratios were highest (0.05-0.15) at very low rates of bile acid excretion, but descended rapidly to a plateau (0.03-0.04) which was constant over the entire range of bile acid excretion rates from 10 to 70 mumoles/min.Similar lipid excretion patterns were observed during glycocholate infusion, but secretin-induced choleresis and dehydrocholate-induced choleresis were unaccompanied by any increments in lecithin or cholesterol excretion and SC2644 (which caused a marked increase in canalicular bile production as measured by erythritol clearance) caused a depression of lipid excretion.The data are consistent with the view that lecithin moves passively from cell membranes to intracanalicular micelles, that transport of cholesterol is coupled to lecithin transport, and that there is also a small amount of independent passive transport of cholesterol from membranes to micelles. A model developed on these assumptions has been shown to behave in a fashion consistent with the entire range of these observations.     

Effects of ion substitution on bile acid-dependent and -independent bile formation by rat liver.

To characterize the transport mechanisms responsible for formation of canalicular bile, we have examined the effects of ion substitution on bile acid-dependent and bile acid-independent bile formation by the isolated perfused rat liver. Complete replacement of perfusate sodium with choline and lithium abolished taurocholate-induced choleresis and reduced biliary taurocholate output by greater than 70%. Partial replacement of perfusate sodium (25 of 128 mM) by choline reduced bile acid-independent bile formation by 30% and replacement of the remaining sodium (103 mM) by choline reduced bile acid-independent bile formation by an additional 64%. In contrast, replacement of the remaining sodium (103 mM) by lithium reduced bile acid-independent bile formation by only an additional 20%, while complete replacement of sodium (128 mM) by lithium reduced bile formation by only 17%, and lithium replaced sodium as the predominant biliary cation. Replacement of perfusate bicarbonate by Tricine, a zwitterionic amino acid buffer, decreased bile acid-independent bile formation by greater than or equal to 50% and decreased biliary bicarbonate output by approximately 60%, regardless of the accompanying cation. In separate experiments, replacement of sodium by lithium essentially abolished Na,K-ATPase activity measured either as ouabain-suppressible ATP hydrolysis in rat liver or kidney homogenates, or as ouabain-suppressible 86Rb uptake by cultured rat hepatocytes. These studies indicate that bile acid(taurocholate)-dependent bile formation by rat liver exhibits a specific requirement for sodium, a finding probably attributable to the role(s) of sodium in hepatic sodium-coupled taurocholate uptake and/or in maintenance of Na,K-ATPase activity. The surprising finding that bile acid-independent bile formation was substantially unaltered by complete replacement of sodium with the permeant cation lithium does not appear to be explained by Na,K-ATPase-mediated lithium transport. Although alternative interpretations exist, this observation is consistent with the hypothesis that much of basal bile acid-independent bile formation is attributable to an ion pump other than Na,K-ATPase, which directly or indirectly mediates bicarbonate transport.     

Hydrocortisone choleresis in the dog

Hydrocortisone sodium succinate (Solu-Cortef; Upjohn Co., Kalamazoo, Mich.) has been found to induce choleresis in unanesthetized fasting dogs fitted with Thomas duodenal cannulae for direct quantitative collection of bile. In all experiments, bile flow increased (average, 68%) 15-20 min after beginning hydrocortisone by infusion in association with an equivalent increment in the output of sodium, potassium, chloride, and bicarbonate. In five animals, the choleretic response occurred independently of, and apparently additive to, the effect of simultaneously administered sodium taurocholate. The fluid added to the bile resembled an ultrafiltrate of plasma. Erythritol clearance increased in proportion to flow, suggesting an effect at the hepatocellular rather than ductal level and probably independent, therefore, of endogenous secretin release. Hydrocortisone and its metabolites were excreted in amounts too small to induce choleresis osmotically. Simultaneous administration of sulfobromophthalein sodium blocked the choleretic response without preventing hydrocortisone excretion. The data suggest that a previously ill-defined mechanism of canalicular bile formation, not mediated by bile salt excretion, may be operative in choleretic response to a variety of agents.     

Sodium Taurocholate Modifies the Bile Acid-Independent Fraction of Canalicular Bile Flow in the Rhesus Monkey

Bile acid-independent secretion and the choleretic response to taurocholate were determined in rhesus monkeys fitted with indwelling silastic cannulas in the common bile ducts. Bile acids were infused intravenously in random order at 3.5, 7.0, or 10.5 mumol/min for 1.5 h each. When data were analyzed with a single regression line, bile flow increased in proportion to the level of bile acid secretion, although the y-intercepts (the conventional measurement of bile acid-independent secretion) varied widely (77.9+/-40.9 ml/24 h). The variation in y-intercepts was observed between animals and with repeated studies in the same animal and could not be explained by sex differences or the effects of the indwelling silastic cannulas, but seemed to be related to the order of bile acid infusion. With only two taurocholic acid infusion rates (7.0 and 3.5 mumol/min), [(14)C]erythritol clearance was greater per mole of secreted bile acid when the initial bile acid infusion was at the high level, but approached zero at low bile acid secretion rates, which suggests that so-called bile acid-independent canalicular flow is closely related to bile acid secretion or is small in size. The augmentation in [(14)C]erythritol clearance when the high infusion rate was given first was also associated with an increase in biliary clearance of [(3)H]inulin, which indicates that the premeability to inulin was also enhanced. Identical experiments which substituted equimolar infusions of a nonmicelle-forming bile acid (taurodehydrocholate) for taurocholate failed to demonstrate any difference in choleretic response or biliary clearance of [(3)H]inulin with the order of bile acid infusion. These experiments demonstrate that a micelleforming bile acid, taurocholate, can increase the permeability of the biliary system to large molecular weight solutes and simultaneously modify the y-intercept and the volume of bile secreted in response to the transported bile acid. Taurocholate may, therefore, modify its own choleretic response, perhaps by altering the structure or function of bile secretory membranes, and appears to be a major determinant of so-called bile acid-independent flow in rhesus monkeys.