Alterations In Intestinal Absorption Research Articles

Comparison of the transport kinetics of fexofenadine and its pH dependency among OATP1A2 genetic variants

Little is known about the influence of non-synonymous genetic variations in the organic anion-transporting polypeptide (OATP) 1A2 on the transport kinetics of its substrate fexofenadine. Moreover, the pH-dependency of fexofenadine uptake also remains unclear. This study aimed to evaluate the effects of genetic variants (Ile13Thr, Asn128Tyr, Glu172Asp, Ala187Thr, and Thr668Ser) on the OATP1A2-mediated uptake of fexofenadine at pH 6.3 and 7.4 and compare the pH dependency of OATP1A2-mediated uptake of fexofenadine and estrone 3-sulfate. The uptake clearances of 0.3 μM and 300 μM fexofenadine were compared with those of 0.3 μM and 300 μM estrone 3-sulfate at pH 6.3 and 7.4. Among the six variants examined, the Thr668Ser variant showed the highest fexofenadine uptake clearance (Vmax/Km); i.e., 4.53- and 6.28-fold higher uptake clearance than the wild type at pH 6.3 and 7.4, respectively. All variants exhibited significantly higher fexofenadine uptake at pH 6.3 than at pH 7.4. Compared with estrone 3-sulfate uptake, the uptake of 0.3 μM fexofenadine was less sensitive to pH. Our findings suggest that genetic variations in OATP1A2 may lead to altered intestinal absorption of fexofenadine, such as increased absorption in subjects bearing the Thr668Ser variant, which showed higher uptake activity.

P-glycoprotein deficiency enhances metabolic activation of and platelet response to clopidogrel through marked up-regulation of Cyp3a11 in mice: Direct evidence for the interplay between P-glycoprotein and Cyp3a

Variability in P-glycoprotein (P-gp) efflux transporting activity was supposed to be involved in altered intestinal absorption and bioavailability of clopidogrel in patients; however, reliable evidence is still lacking. In this study, we sought to determine whether P-gp could play an important role in the metabolic activation of and platelet response to clopidogrel in mice. Abcb1a/1b knock-out (KO) and wild-type (WT) mice were used to evaluate differences in the intracellular accumulation of clopidogrel in the intestine, liver, and brain tissues and in systemic exposure of clopidogrel and its main metabolites as well as the mechanisms involved. Results indicated that, compared with WT mice, KO mice exhibited an 84% increase in systemic exposure of clopidogrel active thiol metabolite H4 and a 14.5% rise of suppression of ADP-induced platelet integrin αIIbβ3 activation, paralleled by a 41% decrease in systemic exposure of clopidogrel due to enhanced systemic clearance. Furthermore, KO mice displayed a 45% increase in Cyp3a11 but a 23% decrease in Ces1 at their protein levels compared with WT mice. Concurrently, intracellular clopidogrel concentrations in the tissues examined did not differ significantly between KO and WT mice. We conclude that although P-gp does not transport clopidogrel and its major metabolites in mice, P-gp-deficient mice exhibit elevated formation of the active metabolite H4 and enhanced antiplatelet effect of clopidogrel through up-regulation of Cyp3a11 and down-regulation of Ces1, suggesting that P-gp activity may correlate inversely with the formation of H4 and antiplatelet efficacy of clopidogrel in clinical settings due to P-gp and CYP3A4 interplay.

Transporter-Mediated Drug-Drug Interactions and Their Significance.

Drug transporters are considered to be determinants of drug disposition and effects/toxicities by affecting the absorption, distribution, and excretion of drugs. Drug transporters are generally divided into solute carrier (SLC) family and ATP binding cassette (ABC) family. Widely studied ABC family transporters include P-glycoprotein (P-GP), breast cancer resistance protein (BCRP), and multidrug resistance proteins (MRPs). SLC family transporters related to drug transport mainly include organic anion-transporting polypeptides (OATPs), organic anion transporters (OATs), organic cation transporters (OCTs), organic cation/carnitine transporters (OCTNs), peptide transporters (PEPTs), and multidrug/toxin extrusions (MATEs). These transporters are often expressed in tissues related to drug disposition, such as the small intestine, liver, and kidney, implicating intestinal absorption of drugs, uptake of drugs into hepatocytes, and renal/bile excretion of drugs. Most of therapeutic drugs are their substrates or inhibitors. When they are comedicated, serious drug-drug interactions (DDIs) may occur due to alterations in intestinal absorption, hepatic uptake, or renal/bile secretion of drugs, leading to enhancement of their activities or toxicities or therapeutic failure. This chapter will illustrate transporter-mediated DDIs (including food drug interaction) in human and their clinical significances.

High stocking density alters bone-related calcium and phosphorus metabolism by changing intestinal absorption in broiler chickens.

Live performance, bone health and metabolic responses to the interaction among stocking density and dietary concentrations of total calcium (TCa) and non-phytate phosphorus (NPP) were determined on 2,232 Ross 308 female broilers over a 3-wk experimental period. From 22 d of age, birds were randomly divided into 48 groups and provided with different corn-soybean meal-based diets varying in TCa (0.70% or 0.90%) and NPP (0.28% or 0.36%) content at 1 of 2 stocking densities [28.6 (LSD, 13 broilers/m2) and 39.6 (HSD, 18 broilers/m2) kg of predicted final BW/m2 floor space], according to a 2 × 2 × 2 factorial design with 6 replications in each treatment. Regardless of NPP supplementation, a high-TCa (0.90%) diet aggravated the impact of HSD on growth (BW gain and feed efficiency, P <0.001) and motility (gait score, P <0.001). This might be explained by deteriorating tibia quality (relative weight, mineral composition and biomechanical property; P <0.01), due to the involvement of decreasing duodenal absorption (type IIb sodium-phosphate co-transporter mRNA, P <0.001) in reduced phosphorus retention (P <0.001). On the contrary, increasing dietary NPP (0.36%), particularly if high in TCa (0.90%), boosted TCa retention (P <0.05) by improving absorption (calcium-binding protein D28k transcription, P <0.05) for LSD chickens, hence enhancing bone development (relative tibia weight and tibia breaking strength, P <0.05) and leg health (walking ability, P <0.05). Together, HSD and LSD birds show an impaired TCa tolerance and a higher TCa+NPP threshold, respectively, to allow the optimization of bone quality via altered intestinal absorption.

Patologías dermatológicas de origen nutricional en los pequeños animales: una revisión

The skin pathologies of nutritional origin either due to alterations in intestinal absorption of some nutrients or genetic predispositions affecting its absorption; nutritional deficiencies may occur by poor development of both home and commercial diets; although at present are not as common for the regulation of food is balanced and demands by control agencies in each country. These conditions are related to some vitamins such as A, D, E and C, essential fatty acids, proteins, and certain minerals such as zinc. Among dermatological diseases and clinical signs include food allergies, dry and oily seborrhea, scaling, follicular plugging, hair loss and dull, dry coat, among others. The aim of this review is closer to the vet to the causes of dermatologic disease of nutritional origin clinical, since alterations of the skin in general are very common in the veterinary clinic for small animals and we need to recognize all the pathologies and treat appropriately. DOI: http://dx.doi.org/10.21615/cesmvz.11.2.7

Inhibition of Matriptase Activity Results in Decreased Intestinal Epithelial Monolayer Integrity In Vitro.

Barrier dysfunction in inflammatory bowel diseases implies enhanced paracellular flux and lowered transepithelial electrical resistance (TER) causing effective invasion of enteropathogens or altered intestinal absorption of toxins and drug compounds. To elucidate the role of matriptase-driven cell surface proteolysis in the maintenance of intestinal barrier function, the 3-amidinophenylalanine-derived matriptase inhibitor, MI-432 was used on porcine IPEC-J2 cell monolayer. Studies with two fluorescent probes revealed that short (2 h) treatment with MI-432 caused an altered distribution of oxidative species between intracellular and extracellular spaces in IPEC-J2 cells. This perturbation was partially compensated when administration of inhibitor continued for up to 48 h. Significant decrease in TER between apical and basolateral compartments of MI-432-treated IPEC-J2 cell monolayers proved that matriptase is one of the key effectors in the maintenance of barrier integrity. Changes in staining pattern of matriptase and in localization of the junctional protein occludin were observed suggesting that inhibition of matriptase by MI-432 can also exert an effect on paracellular gate opening via modulation of tight junctional protein assembly. This study confirms that non-tumorigenic IPEC-J2 cells can be used as an appropriate small intestinal model for the in vitro characterization of matriptase-related effects on intestinal epithelium. These findings demonstrate indirectly that matriptase plays a pivotal role in the development of barrier integrity; thus matriptase dysfunction can facilitate the occurence of leaky gut syndrome observed in intestinal inflammatory diseases.

Alterations in jejunal morphology and serotonin-containing enteroendocrine cells in peripubertal male rats associated with subchronic atrazine exposure

The effect of atrazine on jejunum was investigated by histopathological examination and quantification analysis related to the morphological parameters of the jejunum. The experiment was performed on male Wistar rats from postnatal day 23–51. Atrazine was administered by gavage daily to one group of rats at 50 mg/kg of body weight (bw) dose, to the second at 200 mg/kg bw while the third group was the control. At the end of the experiments after 28 days of treatment, tissue samples were routinely processed and stained with haematoxylin eosin. Additionally, the histochemical staining with periodic acid Schiff-alcian blue was used to demonstrate goblet cells and the immunohistochemistry protocol for serotonin-containing enteroendocrine cells. A significant decrease in the height of epithelial cells covering the intestinal villi, the villus height to total mucosa thickness ratio and the villus height to crypt depth ratio was found at both doses of atrazine. The crypt depth, total mucosa thickness and the thickness of tunica muscularis were significantly increased by the high dose of atrazine only. The number of serotonin-positive endocrine cells significantly decreased in rats treated with the low dose of atrazine. Overall, these results suggest the alterations in intestinal absorption and support the role of atrazine as an endocrine-disrupting compound.

Probiotic lactobacilli: An innovative tool to correct the malabsorption syndrome of vegetarians?

Vegetarians may have subtle nutritional deficiencies which have been related to the occurrence of an unrecognized malabsorption syndrome. The excess phytate content in cereals, nuts, legumes and oilseeds which represent the mainstay of their food intake, seems to play a central role in the pathogenesis of this malabsorption syndrome as an inverse relationship has been shown to link the phytate content of the diet with the intestinal absorption of trace minerals and proteins. We postulate that manipulating the endogenous digestive microflora of subjects on a vegetarian diet through administering probiotic lactic bacteria would represent an innovative tool to counteract the occurrence of the malabsorption syndrome dependent on the high phytate content of their diet. Even though there are no data about the composition of endogenous digestive microflora in subjects on a vegetarian diet, we expect that probiotic lactobacilli can interact with or affect distinct yet interrelated components within the intestinal milieu, such as epithelial cells, enteric flora, and/or mucosal immune cells. This would ultimately translate into the correction of the unregulated mechanisms implicated in the altered intestinal absorption of trace metals and proteins commonly seen in vegetarians. Clinical experience with probiotic therapy of patients with inflammatory bowel disease fully agrees with this view. One additional point of interest is that probiotic lactobacilli, and other species of the endogenous digestive microflora as well, are an important source of the enzyme phytase which catalyses the release of phosphate from phytate and hydrolyses the complexes formed by phytate and metal ions or other cations, rendering them more soluble ultimately improving and facilitating their intestinal absorption. The regular intake of probiotic preparation, may represent a cheap and safe tool in order to convert a diet with a low potential for bioavailability of trace minerals and proteins, such as the vegetarian diet, into a diet with a high bioavailability potential. The benefit of such an approach would not be restricted to vegetarians.

Dietary protein affects intestinal calcium absorption

BackgroundChanges in dietary protein in adults are associated with changes in urinary calcium excretion. The mechanisms underlying this effect are not completely understood, but alterations in intestinal absorption of calcium are not thought to be involved. ObjectiveWe reexamined this mechanism by evaluating the effect of 2 amounts of dietary protein (low: 0.7 g/kg; and high: 2.1 g/kg) on fractional calcium absorption in 7 healthy, young women. DesignThe experiment consisted of 2 wk of a well-balanced diet containing moderate amounts of calcium, sodium, and protein followed by 5 d of an experimental diet that contained 1 of 2 amounts of protein and constant amounts of other nutrients known to influence calcium metabolism. Seven subjects received both amounts of dietary protein in random order. Blood and urine were sampled at baseline and on day 4. Fractional calcium absorption was measured by dual-stable calcium isotopes on day 5. In a second study of 5 additional women, we evaluated the effects of dietary fiber on calcitropic hormones. ResultsSubjects developed hypocalciuria and secondary hyperparathyroidism on day 4 of the low-protein diet. Urinary calcium excretion and the glomerular filtration rate were elevated significantly by day 4 of the high-protein compared with the low-protein diet. Fractional calcium absorption after the low-protein diet was 0.19 ± 0.03, which was significantly lower than that after the high-protein diet (0.26 ± 0.03, P = 0.05). ConclusionThese data provide evidence that depressed intestinal calcium absorption explains, in part, low-protein–induced secondary hyperparathyroidism.

Lymphatic absorption of retinol in young, mature, and old rats: influence of dietary restriction.

Earlier studies have shown that hepatic vitamin A stores increase with age and that rats subjected to life-long dietary restriction (DR) have greater hepatic retinoid concentrations than ad libitum-fed rats. It was suggested that these changes may be attributed to altered intestinal absorption, and so we measured retinol absorption in 3, 12, and 22 month-old female Sprague-Dawley rats fed ad libitum (AL) or a restricted diet (40%) enriched (DR+) or not (DR) with vitamins and minerals. An in vivo model was used and lymph collected from the mesenteric duct while rats were infused a micellar solution containing [3H]retinol into the proximal duodenum. Aging significantly decreased lymph flow, and at all ages DR and DR+ rats exhibited a higher flow than age-matched AL rats. Absorption of retinol was not significantly modified by age in AL and DR+ rats. However, in the DR group, there was a marked decrease in absorption between 3 and 12 months of age. Retinol absorption was significantly higher in 12 and 22 month-old DR+ rats than in age-matched AL rats, a finding that may be explained by both higher lymph flow rate and higher long-term vitamin A intake per body weight. In conclusion, the increased content of vitamin A in the livers of older AL rats is not associated with changes in intestinal absorption. In DR rats, however, retinoid accumulation in the liver may be explained by increased intestinal absorption of the vitamin, though other mechanisms may be involved. A possible explanation may be a decreased output of retinol from the liver as a result of lower metabolic needs. The lower serum vitamin A concentrations observed in DR rats would tend to support this hypothesis.

Triacylglycerol and cholesterol transport during absorption of glycerol trioleate vs. glycerol trielaidate.

We used conscious, chronic lymph-fistula rats to compare intestinal lymphatic transport of glycerol trioleate (TO) vs. glycerol trielaidate (TE) and to determine the effect of TO vs. TE on absorption and transport of cholesterol. Rats were implanted with intestinal lymph fistulas and duodenal cannulas and then given intraduodenal infusions of lipid emulsions containing purified TO or TE (40 mumol/h) and cholesterol (7.8 mumol/h + 2 microCi [14C]cholesterol). Lymph samples were collected at 0, 2, 4, 5, 6, 7, and 8 h after the start of lipid infusion. Lymphatic output and luminal and gut wall recovery of radioactive lipid at 8 h were quantified. Triacylglycerol (TG) fatty acid isomers did not affect lymphatic output of TG; lymph TG fatty acid composition and output reflected infusate composition. Lymphatic output of cholesterol (mass and radioactivity) did not differ between groups; luminal and gut wall recovery of [14C]cholesterol was also similar between groups. Similar lymphatic transport of TG and cholesterol between triolein- and trielaidin-infused rats was maintained for up to 16 h after the cessation of an infused lipid load. These results indicate that TO and TE are transported into lymph similarly, and that cholesterol absorption and transport are similar irrespective of whether TO or TE is the TG source. The data suggest that trans fatty acid-induced hypercholesterolemia is not due to altered intestinal absorption and transport of cholesterol.

Regulation of GLUT5 gene expression in rat intestinal mucosa: regional distribution, circadian rhythm, perinatal development and effect of diabetes

1. GLUT5 gene expression was studied in small intestine under a variety of conditions characterized by altered intestinal absorption of monosaccharides. 2. RNA-blotting studies showed that GLUT5 mRNA was abundantly expressed in rat and rabbit intestine and kidney, but it was not detected in heart or brown adipose tissue. GLUT5 mRNA levels were higher in the upper segments of the small intestine (duodenum and proximal jejunum) than in the lower segments (distal jejunum and ileum). 3. The intestinal expression of GLUT5 mRNA in rat proximal jejunum showed circadian rhythm. A 12-fold increase in GLUT5 mRNA levels was detected at the end of the light cycle and at the beginning of the dark cycle when compared with the early light period. In keeping with this, GLUT5 protein content in brush-border membranes was also increased at the beginning of the dark cycle compared with that in the light period. 4. In streptozotocin-induced diabetes an 80% increase in GLUT5 mRNA levels in mucosa from the proximal jejunum was detected under conditions in which enhanced intestinal absorption of monosaccharides has been reported. 5. The intestinal expression of GLUT5 mRNA showed regulation during perinatal development. Levels of GLUT5 mRNA were low during fetal life, increased progressively during the postnatal period and reached levels comparable with the adult state after weaning. Weaning on to a high-fat diet partially prevented the induction of GLUT5 gene expression. 6. Our results indicate that GLUT5 gene expression is tightly regulated in small intestine. Regulation involves maximal expression in the upper part of the small intestine, circadian rhythm, developmental regulation dependent on the fat and carbohydrate content in the diet at weaning and enhanced expression in streptozotocin-induced diabetes. Furthermore, changes observed in intestinal GLUT5 expression correlate with reported alterations in intestinal absorption of fructose. This suggests a regulatory role for GLUT5 in fructose uptake by absorptive enterocytes.

The effect of long R 3 insulin-like growth factor-I on jejunal glucose absorption in mice

Two experiments were conducted to determine the effects of long R 3 insulin-like growth factor (IGF)-I on jejunal glucose transport in mice. Jejunal rings from 4 mice were incubated for 10 min at 37°C in transport media containing 0, 150 or 500 ng long R 3 IGF-I/ml. Uptake of glucose measured during the last 5 min of incubation was similar among the three treatments. In the second experiment, 8 mice were subcutaneously injected once-daily with 0 or 150 μg long R 3 IGF-I/kg BW for 3 d. Treatment with long R 3 IGF-I did not affect jejunal glucose absorption. Weight of the stomach and small and large intestine were similar for both groups, however, treatment with long R 3 IGF-I reduced the length of the small intestine (P<0.05). There was also a tendency for jejunal villus height (P<0.1) and thickness of the muscle layers (P<0.11) to be reduced by long R 3 IGF-I. Short-term administration of long R 3 IGF-I does not appear to cause alterations in intestinal absorption of glucose.

Isolation, characterization, and developmental expression of pig intestinal fatty acid-binding proteins

The goal of this study was to characterize and quantify intestinal fatty acid-binding proteins of the pig. Small intestinal mucosa from 13–19 kg pigs was homogenized and centrifuged to obtain cytosol. Isolation of fatty acid-binding proteins from delipidated cytosol was achieved using molecular sieve, oleic acid affinity, and ion exchange chromatography. Fatty acid-binding protein isolation was monitored using a fatty-acid binding assay in conjunction with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. Antisera to rat liver-fatty acid-binding protein cross reacted with an isolated intestinal fatty acid-binding protein of M r = 13,000, whereas antisera to rat intestine-fatty acid-binding protein was not cross reactive with isolated pig intestinal proteins. These experiments identify a pig intestinal fatty acid-binding protein that exhibits strong immunochemical similarity to rat liver-fatty acid-binding protein. Cytosol prepared from intestinal mucosa of pigs at −4, 2, 4, 7, 15, 22, 28, and 35 d of age was assayed for fatty acid-binding protein activity. Preweaning fatty acid-binding protein activity in cytosol was maximal at 7 days of age when expressed as total jejunal fatty acid binding per kilogram bodyweight, intestinal or mucosal weight or milligram total protein. After weaning (21 d), fatty acid-binding protein activities declined to 28 days, but increased again by 35 days. Total soluble fatty acid-binding protein activity in pig intestine is regulated during postnatal development and this may account in part for the altered intestinal absorption of lipids observed in young pigs at weaning.

Investigation of intestine function during acute viral hepatitis using combined sugar oral loads.

One fifth of all cases of A virus hepatitis (AVH) have symptoms of gastroenteritis at the onset. This study investigated the mediated intestinal absorption of D-xylose (D-xyl) and 3-o-methyl-D-glucose (3-omG) and the non-mediated permeation of lactulose (Lacl, mol wt 342) and L-rhamnose (L-rh, mol wt 164) during acute and remission phases of AVH. Ten patients with AVH were given an oral load containing these sugars (5 g D-xyl: 2.5 g 3-omG, 1 g L-rh, 5 g lacl in 250 ml water) once during the acute phase and again during remission. The same load was given once to a group of 22 healthy controls. The mean concentration of D-xyl in urine and the ratio of D-xyl to 3-omG in plasma and urine were normal in both the AVH phases, ruling out intestinal malabsorption even in the acute phase. This study showed a significant increase in non-mediated permeation to Lacl, but not to L-rh, during the acute phase. These data indicate that the barrier function of the intestine is compromised in AVH infection while the absorptive function is not. An abnormally low concentration of D-xyl and 3-omG in plasma at one hour was found in all patients during the acute phase. This finding cannot be explained by alterations in intestinal absorption, but could be accounted for by increased space distribution of the sugars because of increased diffusion into tissue cells and/or expansion of the extracellular space by fluid retention.

Impaired intestinal absorption of riboflavin in experimental uremia.

Increased plasma and red blood cell concentrations of riboflavin have been reported in uremia. The possible role of altered intestinal absorption of riboflavin in the genesis of this abnormality is not known. For this reason we examined the intestinal absorption of riboflavin in rats made uremic by subtotal nephrectomy and sham-operated (control) rats in vivo using the recycling perfusion technique and in vitro using the everted-sac technique. Paradoxically, the results showed a significant impairment of intestinal absorption of riboflavin in vivo in uremic rats compared to the control group. However, no significant difference was observed in riboflavin transport in vitro. We conclude that the intestinal absorption of riboflavin is decreased in experimental uremia and cannot account for the reported increase in its plasma and red blood cell concentrations.

Alcohol and the Gastrointestinal Tract

The effects of acute and chronic ethanol ingestion on esophageal motility and the potential complications of these alterations are reviewed. Injury to the gastric mucosa and the small intestine and alterations in intestinal absorption can also result from alcohol abuse.

Comparative effects of diethylhexyl phthalate or monoethylhexyl phthalate on male mouse and rat reproductive organs

Sexually mature mice injected (ip or sc) with doses ranging from 1 to 100 mg/kg monoethylhexyl phthalate (MEHP) or 50 to 100 mg/kg diethylhexyl phthalate failed to exhibit any significant alterations in testicular weight or zinc levels. Further, no changes in mouse seminal vesicle or anterior prostate weight and zinc levels occurred. Rats injected with MEHP (50 mg/kg) showed a 37% decrease in prostatic zinc; DEHP (100 mg/kg) caused a 33% decrease in prostatic zinc and a 31% decrease in gonadal zinc. These studies indicated that the reproductive system of the male rat is more sensitive to phthalate-induced toxicities than the mouse. Finally, phthalate acid ester-induced gonadal zinc depletion was evident despite the fact that the MEHP or DEHP was administered ip or sc, thus apparently eliminating altered intestinal absorption as the cause for the species differences.

Absorption of d-glucose by the rabbit jejunum during cholera toxin-induced diarrhoea.

CHOLERA exotoxin causes fluid production by the intestine when instilled into jejunal or ileal loops of anaesthetized rabbits1–3. Profuse quantities of isotonic fluid are produced by the small intestine without histological alteration of the mucosa4. The pathogenesis of the increased fluid production by the intestine in cholera is unsettled. It has not been determined whether the fluid production is the result of increased intestinal secretion, decreased intestinal absorption or a combination of these two factors. To assess the importance of altered intestinal absorption, absorption of d-glucose by the rabbit jejunum was measured both in the presence and absence of cholera diarrhoea.