Intestinal handling of a glucose gavage by the rat

An oral gavage of either 3, 1 or 0.1 mmoles of ¹⁴C-labelled glucose was given to rats under standard feeding conditions or food deprived for 24 hr. The fate of the glucose label was determined at 10, 15, 30 and 60 min after gavage; at 60 min 40% of the glucose was absorbed in fed rats (60% in food deprived). The portal vein blood flows were determined and the levels of glucose, lactate, alanine and pyruvate, and their radioactivity, as well as that of CO, were measured in both portal and arterial blood.The net computed glucose and 3-carbon carriers (lactate, alanine and pyruvate) actually released into the portal system by the intestine was lower than the amount of glucose taken up from the intestinal lumen in one hour. Oxidation to ¹⁴CO₂ accounted for a 12–15% of the absorbed glucose. The size of the gavage deeply affected the proportion of glucose released into the portal blood (c. 50% with a 3 mmoles gavage and practically nil with a 0.1 mmoles gavage), but it affected much less the generation of lactate and other 3 C carriers. In fed rats, the net intestinal balance of non-radioactive glucose was negative, and that of lactate positive; when radioactive glucose was considered, the pattern was inverted. In starved rats, both glucose and lactate were released in large proportions by the intestine, but alanine efflux was lower.It can be concluded that the intestine consumes a considerable proportion of glucose in the fed state. Glucose handling by the intestine is compartmentalized in two functional circuits: glucose is taken up from the arterial blood and used for intestinal metabolism and lactate production, luminal glucose is absorbed mainly unaltered and transferred to the portal blood. Thus, the generation of lactate is mainly related to the availability of arterial glucose. In addition to the release of the ingested glucose as 3 C carriers or glucose, an extraportal pathway for glucose transfer into the bloodstream is postulated.     

Absorption of a protein gavage in Zucker lean rats. Influence of protein content in the diet.

The rate of protein absorption was measured in Zucker lean rats. Rats were fed with a bolus that contained ca. 300 mg of 14C-labelled protein at the beginning of the light cycle. Blood was extracted from the portal vein at intervals up to 9 hours after gavage. Label incorporation into tissue protein was monitored. The digestion and absorption of protein was slow, and 9 hours after the gavage, 20% of the bolus remained in the stomach. Forty percent of the protein was absorbed in the first hour. This was followed first by a linear absorption process, then by the amino acid incorporation into tissue proteins. The appearance of label in the portal vein increased progressively for up to four hours, shifting to a progressive decrease that coincides with the maintenance of this label in the tissues. The skin, the striated muscle and the liver showed the highest amounts of labelled proteins. The application of this model to animals fed low-(LP) or high-protein (HP) content diets showed that the HP group digested the protein faster than the LP group, and that catabolism of the amino acids was higher in the HP group. The LP group digested protein much more slowly than the RD (control) group, but protein accretion was more efficient.     

Rat liver amino acid balances after the administration of an oral protein load.

The hepatic balances of amino acids, ammonia and urea have been measured in rats for three hours after receiving a protein load. The liver took up practically all of the portal ammonium. Alanine was retained to a large extent during all three hours. Other portal amino acids, mainly essential amino acids, were largely retained in about one hour after the gavage, to be released in a similar proportion thereafter. The other amino acids were also retained and then released, but to a lower extent. These amino acids were used in part by the liver for the synthesis and release of urea, which appearance in hepatic vein peaked at two hours after the protein administration.     

Basolateral aromatic amino acid transporter TAT1 (Slc16a10) functions as an efflux pathway

Basolateral efflux is a necessary step in transepithelial (re)absorption of amino acids from small intestine and kidney proximal tubule. The best characterized basolateral amino acid transporters are y+LAT1-4F2hc and LAT2-4F2hc that function as obligatory exchangers and thus, do not contribute to net amino acid (re)absorption. The aromatic amino acid transporter TAT1 was shown previously to localize basolaterally in rat's small intestine and to mediate the efflux of L-Trp in the absence of exchange substrate, upon expression in Xenopus oocytes. We compared here the amino acid influx and efflux via mouse TAT1 in Xenopus oocytes. The results show that mTAT1 functions as facilitated diffusion pathway for aromatic amino acids and that its properties are symmetrical in terms of selectivity and apparent affinity. We show by real-time RT-PCR that its mRNA is highly expressed in mouse small intestine mucosa, kidney, liver, and skeletal muscle as well as present in all other tested tissues. We show that mTAT1 is not N-glycosylated and that it localizes to the mouse kidney proximal tubule. This expression is characterized by an axial gradient similar to that of the luminal neutral amino acid transporter B0AT1 and shows the same basolateral localization as 4F2hc. mTAT1 also localizes to the basolateral membrane of small intestine enterocytes and to the sinusoidal side of perivenous hepatocytes. In summary, we show that TAT1 is a basolateral epithelial transporter and that it can function as a net efflux pathway for aromatic amino acids. We propose that it, thereby, may supply parallel exchangers with recycling uptake substrates that could drive the efflux of other amino acids.     

Disposition and metabolism of 2-fluoro-β-alanine conjugates of bile acids following secretion into bile

Since 2-fluoro-β-alanine (FBAL) conjugates of bile acids (BA), the primary biliary metabolites of fluoropyrimidine (FP) drugs, have been suggested to be related to the hepatotoxicity which develops in patients receiving FP chemotherapy by intrahepatic arterial infusion (Proc. Natl. Acad. Sci. USA 84, 5439–5443, 1987), it was important to determine whether they undergo enterohepatic circulation and hence accumulate in the liver and biliary system. In initial studies, sensitivity of FBAL-BA conjugates to hydrolysis by pancreatic enzymes was examined. In subsequent in vivo studies, a model FBAL-BA conjugate, FBAL-chenodeoxycholate (FBAL-CDC), was introduced into the lumen of the small intestine of anesthetized rats with biliary fistulas to quantitate the intestinal absorption, metabolism and tissue distribution of the conjugate. The results indicated that: (1) FBAL-BA conjugates were resistant to hydrolysis by pancreatic enzymes (carboxypeptidase A, carboxypeptidase B and trypsin) and by human pancreatic juice, but were completely hydrolyzed by cholylglycine hydrolase. (2) At least one-half of the administered FBAL-CDC was deconjugeted during the process of intestinal absorption, as shown by HPLC analysis of the radioactivity in portal venous blood. (3) Deconjugated FBAL or CDC was reconjugated in liver with other bile acids or amino acids (glycine and taurine), respectively, as shown by radiochromatography of bile. (4) FBAL, formed as a result of hydrolysis of FBAL-CDC, had a wide tissue distribution. In conclusion, FBAL-CDC has a rapid turnover during its enterohepatic circulation due to deconjugation in the intestine and reconjugation in the liver.     

Disposition and metabolism of 2-fluoro-beta-alanine conjugates of bile acids following secretion into bile

Since 2-fluoro-beta-alanine (FBAL) conjugates of bile acids (BA), the primary biliary metabolites of fluoropyrimidine (FP) drugs, have been suggested to be related to the hepatotoxicity which develops in patients receiving FP chemotherapy by intrahepatic arterial infusion (Proc. Natl. Acad. Sci. USA 84, 5439-5443, 1987), it was important to determine whether they undergo enterohepatic circulation and hence accumulate in the liver and biliary system. In initial studies, sensitivity of FBAL-BA conjugates to hydrolysis by pancreatic enzymes was examined. In subsequent in vivo studies, a model FBAL-BA conjugate, FBAL-chenodeoxycholate (FBAL-CDC), was introduced into the lumen of the small intestine of anesthetized rats with biliary fistulas to quantitate the intestinal absorption, metabolism and tissue distribution of the conjugate. The results indicated that: (1) FBAL-BA conjugates were resistant to hydrolysis by pancreatic enzymes (carboxypeptidase A, carboxypeptidase B and trypsin) and by human pancreatic juice, but were completely hydrolyzed by cholyglycine hydrolase. (2) At least one-half of the administered FBAL-CDC was deconjugated during the process of intestinal absorption, as shown by HPLC analysis of the radioactivity in portal venous blood. (3) Deconjugated FBAL or CDC was reconjugated in liver with other bile acids or amino acids (glycine and taurine), respectively, as shown by radiochromatography of bile. (4) FBAL, formed as a result of hydrolysis of FBAL-CDC, had a wide tissue distribution. In conclusion, FBAL-CDC has a rapid turnover during its enterohepatic circulation due to deconjugation in the intestine and reconjugation in the liver.     

Plasma levels and renal handling of endogenous amino acids in snakes: a comparative study

Plasma levels of 22 endogenous amino acids were measured by ion-exchange chromatography in four species of snakes: Thamnophis sirtalis, T. radix, Aipysurus laevis, and Python molurus. Despite considerable interspecific variation in the amino acid composition, all species showed relatively high plasma concentrations of histidine, a feature apparently unique to reptiles. The renal handling of these amino acids was studied by renal clearance methods. As in other vertebrates, net tubular absorption of filtered amino acids predominated. However, net tubular secretion of taurine, cysteic acid and/or phosphoserine and beta-alanine was observed, with taurine being the predominant amino acid secreted. The percentage reabsorption of the total amino acids filtered by the snake kidneys ranged from 79 to 95%. Evidence for the postrenal absorption of amino acids in these reptiles is presented. In species that normally undergo hibernation (Thamnophis spp.), the ability of the kidney to reabsorb amino acids was depressed by cold acclimation. Cold acclimation significantly decreased plasma levels of all amino acids except taurine, whose concentration increased. The increase in plasma taurine level may have resulted from cellular osmoregulation. Under these conditions, renal excretion of taurine increased concomitantly with the increase in plasma taurine concentration.     

河蚬汤中氨基酸组分的分析

利用全自动氨基酸分析仪测定了传统保肝食品--河蚬汤中的总氨基酸和游离氨基酸含量.结果表明,河蚬汤中含有17种常见(色氨酸在酸解时遭破坏)氨基酸,还有鸟氨酸和牛磺酸两种非蛋白质组成氨基酸,总含量为297.4 mg·g-1.其中鸟氨酸含量最高,占总氨基酸的12.4%,且87%的鸟氨酸以结合态形式存在.河蚬汤中游离氨基酸含量...     

Taurine transport by the flounder (Pseudopleuronectes americanus) intestine

Taurine transport by the flounder gut was characterized in isolated strips of intestine mounted between Ringer's solutions. Taurine was transported into the cell, against its concentration gradient, by a sodium-dependent system present in both the mucosal and serosal membranes. This system appears to be specific for beta-amino acids and is regulated by cyclic nucleotides (cGMP and cAMP). Kinetic analyses indicated that under physiological conditions the magnitudes of the bidirectional taurine uptake rates would favor net absorption of taurine from lumen to blood.     

Intestinal handling of an oral oleoyl-estrone gavage by the rat

Adult Zucker lean (Fa/?) female rats received a single 250 nmol oral gavage of 3H-labelled oleoylestrone in 0.2 ml of sunflower oil. After one hour, samples of arterial, portal and suprahepatic blood, and lymph were obtained and fractioned to determine the amount of radioactivity present in the form of free estrone, acyl-estrone and hydrophilic estrone esters in the blood of each vessel. Lipoprotein fractions (chylomicra + VLDL, LDL, HDL and lipoprotein-depleted plasma) were also analysed as well as the distribution of absorbed 3H-estrone in the intestine, specific organs and carcass. About one third of the oleoyl-estrone dose recovered was found in the tissues, mainly in the blood, the rest remaining relatively untouched in the intestinal content. High hypothalamic estrone uptake (compared with the rest of the brain) was observed. Data from non-radioactive estrone measurements showed a similar pattern of absorption and tissue distribution to that obtained by 3H-estrone tracking alone. In both cases, most of the estrone present in the intestinal lumen was absorbed as intact oleoyl-estrone, but a significant part was absorbed as free estrone. There is a net transfer of 3H-estrone into portal blood HDL, and part of the 3H-estrone is also loaded into lymph-carried chylomicra. A large share of free estrone is filtered by the liver, but most of the acyl-estrone absorbed passes unaltered. The oral administration of oleoyl-estrone results in significant absorption of the unaltered molecule, which is transferred to lymph-carried chylomicra and also directly to plasma HDLs. It may be inferred that the HDL fraction contains the physiological carrier of oleoyl-estrone in its role of ponderostat signal.     

Some pathomorphological bases for the disorders of the tissue amino acid pool in Salmonella infection]

Possible causes of disturbances of the amino acid pool of the blood serum and the tissue, as well as the presence of an association between these shifts and the changes in the ultrastructure of the striated columnar epithelium of the small intestine were studied. Investigations were conducted in 138 patients suffereing from salmonellosis and in 120 albino rats experimentally-infected with S. typhimurium culture. Free amino acids of the blood serum and the tissue were determined by paper chromatography and on the automatic amino acid analyzer; ultrastructure of the small intestinal mucosa was studied by electron microscopy. The height of salmonella infection was attended by dysproportional changes in the amino acid pool of the tissue and the blood caused by disturbance of absorption and intracellular transformation of amino acids. Ultrastructural changes in the microvilli of the absorbing cells of the small intestine underlied absorption disturbances: at the height of salmonella infection some of the microvilli of the striated columnar small intestine proved to show poor contours; reduction of mitochondrial cristae was observed in almost all the mitochondria.     

Effects of prolonged guanidinoethanesulphonate administration on taurine and other amino acids in rat tissues

In order to deplete tissue taurine, 2-guanidinoethanesulphonate, a structural analogue of taurine was administered in drinking water with taurine-free diet to adult rats for four weeks. As a consequence the taurine concentrations in the blood serum, liver, kidney, spleen, intestine, lung, heart, muscle and cerebellum fell by nearly one half. Threonine, serine, glycine, alanine, methionine, tyrosine, lysine and histidine concentrations increased in blood plasma. Similar changes were also discernible in the heart and muscle. In the kidney and the lung the concentrations of several other amino acids fell as well, though increments occurred in the threonine content in the kidney and in threonine, serine and methionine contents in the lung. Taurine was practically the only amino acid the level of which fell in the liver, spleen, intestine and cerebellum. These findings indicate that 2-guanidinoethanesulphonate combined with taurine-free diet effectively lowers tissue taurine levels, but its action is not specific to taurine. It may be used as a tool to elucidate the physiological functions of taurine in the body.     

Amino acid transport in the intestine of the caiman

Seventeen amino acids were fed singly to small caimans and the rates of their disappearance from the gut lumen, and of their appearance in intestinal mucosa, whole intestine, whole stomach, and plasma were determined. The results were compared with those in which massive amounts of protein were fed. When single amino acids were fed, only traces of arginine, ornithine, lysine, aspartate and asparagine were absorbed intact. Glycine, alanine and serine were absorbed rapidly reaching mucosal concentrations as high as 40 mM. The others were not concentrated as highly and most were absorbed by the mucosa more slowly than the glycine group. Protein feeding did not result in high amino acid concentrations in the mucosa. Whether amino acids were ingested as protein or in the free state, glycine, alanine and glutamine increased in the mucosa, suggesting these three incorporate nitrogen released from the others. It appeared that several transport systems operate if amino acids are given singly, and that a different more efficient transport system operates during protein digestion.     

Metabolism of absorbed aspartate,asparagine, and arginine by rat small intestine in vivo

l-[U-¹⁴C]aspartate, l-[U-¹⁴C]asparagine, and l-[U-¹⁴C]arginine were administered luminally into isolated segments of rat jejunum in situ, and the radioactive products appearing in venous blood from the segment were identified and quantified, in a continuation of similar studies with l-glutamate and l-glutamine (Windmueller H.G. and Spaeth, A. E. (1975) Arch. Biochem. Biophys. 171, 662–672). Aspartate, administered alone (6 mm) or with 18 other amino acids plus glucose, was absorbed more rapidly than glutamate, but, as with glutamate, less than 1% was recovered intact in intestinal venous blood. More than 50% of aspartate carbon was recovered in CO₂, 24% in organic acids, mostly lactate, 12% in other amino acids (alanine, glutamate, proline, ornithine, and citrulline), and 10% in glucose, apparently the first demonstration of gluconeogenesis by intestine in vivo. In contrast to aspartate and glutamine, nearly all asparagine was absorbed intact, less than 1% being catabolized. About 4% of the absorbed dose was incorporated into the acid-insoluble fraction of intestine, as was the case with all the amino acids studied. In conventional or germ-free rats, only 60% of arginine was absorbed intact, while 33% was hydrolyzed to ornithine and urea. The urea and 38% of the ornithine were released into the blood; the remaining ornithine was metabolized further by intestine to citrulline, proline, glutamate, organic acids, and CO₂. Catabolism of several amino acids from the lumen plus glutamine from arterial blood may provide an important energy source in small intestine.     

Effects of 2-hydroxy-4-methylthiobutyrate on portal plasma flow and net portal appearance of amino acids in piglets

To determine whether portal plasma flow (PPF) and net portal appearance of amino acids (AA) could be affected by 2-hydroxy-4-methylthiobutyrate (HMB), six barrows (35-day-old, 8.6 ± 1.4 kg), implanted with arterial, portal and mesenteric catheters, were fed a dl-methionine (as the control) or HMB-supplemented diet once hourly and infused intramesenterically with 1% p-amino hippurate. PPF was numerically 9% higher (P = 0.09) in HMB-fed pigs than in controls over a 4–6 h period. Compared with controls, pigs fed the HMB diet had increased (P < 0.05) net portal balance and/or appearance of leucine, isoleucine, histidine, arginine and alanine, but had decreased (P < 0.05) portal appearance of glutamate over a 6-h period. The concentration of acetate in the lumen of the distal small intestine was higher (P = 0.01) in HMB-fed pigs than in controls (25.14 vs. 7.64 mmol/kg). mRNA levels for proglucagon and endothelial nitric-oxide synthase (eNOS) in stomach and proximal small intestine, and mRNA levels for GLP-2 receptor (GLP-2R) in stomach were higher (P < 0.05) in HMB-fed pigs compared with those in controls. Collectively, HMB supplementation increased concentrations of short-chain fatty acids in intestinal lumen, expression of proglucagon, GLP-2R, and eNOS genes, and net portal absorption of AA. These novel findings from the study with pigs may also have important implications for intestinal nutrition and health in humans.     

Glucagon-like peptide-2 (GLP-2) increases net amino acid utilization by the portal-drained viscera of ruminating calves

Glucagon-like peptide-2 (GLP-2) increases small intestinal mass and blood flow in ruminant calves, but its impact on nutrient metabolism across the portal-drained viscera (PDV) and liver is unknown. Eight Holstein calves with catheters in the carotid artery, mesenteric vein, portal vein and hepatic vein were paired by age and randomly assigned to control (0.5% bovine serum albumin in saline; n = 4) or GLP-2 (100 μg/kg BW per day bovine GLP-2 in bovine serum albumin; n = 4). Treatments were administered subcutaneously every 12 h for 10 days. Blood flow was measured on days 0 and 10 and included 3 periods: baseline (saline infusion), treatment (infusion of bovine serum albumin or 3.76 μg/kg BW per h GLP-2) and recovery (saline infusion). Arterial concentrations and net PDV, hepatic and total splanchnic fluxes of glucose, lactate, glutamate, glutamine, β-hydroxybutyrate and urea-N were measured on days 0 and 10. Arterial concentrations and net fluxes of all amino acids and glucose metabolism using continuous intravenous infusion of [U¹³-C]glucose were measured on day 10 only. A 1-h infusion of GLP-2 increased blood flow in the portal and hepatic veins when administered to calves not previously exposed to exogenous GLP-2, but after a 10-day administration of GLP-2 the blood flow response to the 1-h GLP-2 infusion was substantially attenuated. The 1-h GLP-2 infusion also did not appreciably alter nutrient fluxes on either day 0 or 10. In contrast, long-term GLP-2 administration reduced arterial concentrations and net PDV flux of many essential and non-essential amino acids. Despite the significant alterations in amino acid metabolism, glucose irreversible loss and utilization by PDV and non-PDV tissues were not affected by GLP-2. Fluxes of amino acids across the PDV were generally reduced by GLP-2, potentially by increased small intestinal epithelial growth and thus energy and amino acid requirements of this tissue. Increased PDV extraction of glutamine and alterations in PDV metabolism of arginine, ornithine and citrulline support the concept that GLP-2 influences intestine-specific amino acid metabolism. Alterations in amino acid metabolism but unchanged glucose metabolism suggests that the growth effects induced by GLP-2 in ruminants increase reliance on amino acids preferentially over glucose. Thus, GLP-2 increases PDV utilization of amino acids, but not glucose, concurrent with stimulated growth of the small intestinal epithelium in post-absorptive ruminant calves.     

Arteriovenous differences for amino acids across control and acid-secreting rat stomach in vivo.

1. A method is described for measuring arteriovenous differences across the rat stomach in vivo. 2. Notable results were the uptake of branched-chain amino acids, the uptake of arginine, which was approximately balanced by an output of ornithine, and the output of alanine. 3. The fractional extraction of glutamine from the blood by the stomach wall of pentagastrin-stimulated rats was 4.7%. 4. The arteriovenous differences for ammonia depended upon the blood ammonia concentration. 5. Arteriovenous differences were not affected by the stimulation of acid secretion with pentagastrin. 6. It is concluded that the high activity of branched-chain-amino-acid aminotransferase (EC 2.6.1.42) in the gastric mucosa is associated with metabolism of these amino acids, but that the stomach wall is a less avid user of glutamine than is the small intestine.