Marwan Farouk

Localization and characterization of secretin binding sites expressed by rat bile duct epithelium

The goal of the present studies was to identify and characterize the site of secretin action in the liver. Sections of normal and bile duct-ligated rat livers were used for in vitro 125I-secretin receptor autoradiography. Saturable binding was observed in both normal and bile duct-ligated livers but was much greater in the bile duct-ligated preparations. Binding was limited to biliary epithelium and the increased secretin binding observed in the ligated livers correlated with the increase in ductular tissue. Saturable binding was inhibited in a dose-dependent fashion by increasing concentrations of nonradioactive secretin. Analysis of saturation binding showed that 125I-secretin binding was best fit by a one-site receptor model with a Kd of 5.3 +/- 1.1 nmol/L. Glucagon, vasoactive intestinal polypeptide, gastric inhibitory polypeptide, growth hormone-releasing hormone, and cholecystokinin did not inhibit saturable 125I-secretin binding at concentrations of 1 pmol/L to 1 mumol/L. The authors conclude that high-affinity, specific secretin binding sites are present in rat intrahepatic biliary epithelium. When bile ducts are stimulated to proliferate by bile duct ligation, secretin binding is also increased.

Intracerebroventricular neuropeptide Y stimulates bile secretion via a vagal mechanism.

The effect of intracerebroventricular injection of neuropeptide Y on biliary secretion was studied in conscious dogs, prepared with gastric and duodenal fistulas and cerebroventricular guides. Bile secretion was increased in a dose-dependent fashion by intracerebroventricular neuropeptide Y. The peak increase was seen after 500 pM/kg of neuropeptide Y which resulted in a 30 x 2% increase in bile flow over the period 30-150 minutes after injection. (Control: 23 x 2 (1 x 2) ml/2 hours; neuropeptide Y 500 pM/kg: 30 x 5 (1 x 1) ml/2 hours). Biliary lipid composition was not altered significantly but bicarbonate output was increased at all doses tested. Intravenous infusion of neuropeptide Y (1000 pM) for 1 hour had no significant effect. Intracerebroventricular neuropeptide Y (1000 pM/250-300 mg body weight) also increased bile flow in urethane-anaesthetised rats. This effect was abolished by cervical vagotomy. The demonstration of a central stimulation of alkaline bile flow suggests that bile secretion may be subject to central modulation.

Technique of intraluminal biliary ultrasonography during laparoscopic cholecystectomy

Intraluminal ultrasonography of the common duct was performed in nine patients undergoing laparoscopic cholecystectomy, using a system comprising a 20-MHz crystal in a 95-cm, blunt-tipped 6F sheath, mechanically rotated at 1,800 rpm. The probe was introduced through an incision in the cystic duct and passed into the duodenum. When the catheter was withdrawn, excellent visualization of the common and cystic ducts and lower end of the common hepatic duct was achieved. In seven patients, the biliary tree was normal. A small calculus was discovered in the common duct in one patient. This stone was not seen on a subsequent cholangiogram and was subsequently retrieved. An additional patient had mucus or sludge noted in the duct, which cast no acoustic shadow and thus was distinguished from calculi. The technique was fast, efficient, and easy to perform in this small group of patients and holds promise for screening the common duct pathology during laparoscopic cholecystectomy.

The effects of liver denervation on the regulation of hepatic biliary secretion.

Effects of liver denervation on bile formation were studied in eight dogs prepared with chronic biliary fistulas. The animals were studied in the basal state, after feeding, and during infusion of glucagon 50 ng/kg/min, secretin 2 U/kg/hr, or somatostatin 200 ng/kg/min. After this first set of experiments the animals underwent a total hepatic denervation that consisted of section of the hepatic ligaments and a careful dissection of the portal vein, hepatic artery, and common duct with stripping of all the surrounding connective tissue and topical application of phenol. The above experiments were then repeated. Denervation did not modify bile flow, or bile salts, cholesterol, or phospholipid concentration or output. Biliary response to glucagon and secretin was similar before and after denervation. Somatostatin had an anticholerectic effect in both intact and denervated animals, but significantly reduced bile salt output only in the intact dogs. Feeding had a choleretic effect pre- and postdenervation, and the infusion of somatostatin following feeding decreased bile flow to the same degree before and after denervation. In the intact animals the output of all three biliary lipids was reduced by somatostatin after feeding but they were unaffected by somatostatin after denervation. Moreover, cholesterol and phospholipid outputs were stable after feeding in intact animals, but significantly decreased after denervation. 14C-erythritol clearance studies indicated no change in the canalicular component of bile flow with denervation, except again during somatostatin suppression of feeding. These data indicate that basal bile flow is normal after denervation but that innervation may play an important role in the modulation of responses to somatostatin and more complex stimuli such as feeding.

Biliary response to glucagon in humans.

Glucagon has been demonstrated to have profound effect on biliary secretion in several species. Glucagons biliary effects were studied in humans following biliary tract surgery. Nine patients underwent common bile duct exploration and insertion of a balloon-occludable t tube. An aliquot of the collected sample was kept and the enterohepatic circulation was maintained by reinfusion of the collected bile via the distal t-tube port. Glucagon increased bile flow and decreased cholesterol and phospholipid output during stable bile acid output. Furthermore high-performance liquid Chromatographie analysis of bile acid profiles revealed no significant changes in bile salt species or conjugation after glucagon infusion. Glucagon is probably important in the physiologic regulation of biliary secretion in humans.

Exogenous Neuropeptide Y Blocks Myoelectric Activity in the Upper Gastrointestinal Tract of Starved Dogs: Brain Neuropeptide Y Converts a Fasting Pattern of Myoelectric Activity to a Fed Pattern

The effect of intracerebroventricular (ICV) neuropeptide Y (NPY) on the migrating motor complex (MMC) was examined in five starved dogs. Myoelectric activity was recorded using gastric, duodenal, and jejunal electrodes. Intragastric pressure was monitored via a gastric fistula, and ICV injections were given through a cerebroventricular guide. Recordings were made with no ICV injection and before and after 250-microliters bolus injections of vehicle as control (0.1% dog serum albumin in saline) or 500 pmol/kg NPY. The mean interval between MMCs was 98 +/- 10 min without ICV injection and 96 +/- 7 min after control solution. After ICV injection of NPY no further MMCs were recorded in any dog, even though the study was continued for a minimum of 3.5 h. Instead, the myoelectric pattern became indistinguishable from that in fed dogs. We conclude that central NPY plays a role in modulation of upper gastrointestinal myoelectric activity. This may reflect a central regulatory role for NPY in the coordination of feeding.

Cholesterol secretion from hepatocytes induced by triacylglycerol and apolipoprotein E

The mechanism for the increase in plasma cholesterol levels in cholesterol-fed rats following chylomicron transport was investigated in intact animals, in isolated perfused liver, and in hepatocytes in monolayer cultures. Intravenous administration of egg phosphatidylcholine in amounts greater than those required to cause a plasma cholesterol response when given as chylomicrons was without effect. This makes it unlikely that increased plasma cholesterol levels resulted from the recruitment of tissue cholesterol by the plasma chylomicron phospholipids that persisted in the plasma after triacylglycerol clearance. The hepatic origin of the increased plasma cholesterol levels was directly confirmed by two hepatic perfusion experiments. When cholesterol-fed rats received intravenous chylomicrons prior to isolated hepatic perfusion, more cholesterol was secreted by the liver than when the rats were injected intravenously with buffer. Perfusion of apolipoprotein E (apo E)-rich triacylglycerol emulsions through the livers also enhanced cholesterol secretion. The increase in hepatocyte cholesterol secretion seen with cholesterol-fed rats was also noted in monolayer cultures following incubation with apo E rich-triacylglycerol emulsions. The apolipoprotein or the emulsion alone, or apo E-rich phosphatidylcholine liposomes, had no effect. The data confirm previous indirect observations that the liver is the source of cholesterol that appears in plasma following transport of chylomicrons or following a lipid-rich meal in cholesterol-fed rats. The data also re-emphasize the importance of providing apo E with triacylglycerol emulsions to initiate secretion of lower density lipoproteins by the liver.