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Objective: To evaluate the transepithelial transport of sodium, glucose, potassium, and water and the mRNA level of the sodium–glucose cotransporter (SGLT1) and the facilitated sugar transporter (GLUT2) in the small intestine of iron-deficient rats. Methods: After 6 wk of receiving diets with low or normal iron content, rats (Wistar-EPM) were subjected to two experiments: 1) evaluation of the transepithelial transport of sodium, glucose, potassium, and water by an “in vivo” experimental model of intestinal perfusion and 2) determination of relative SGLT1 and GLUT2 mRNA levels in the proximal, intermediate, and distal portions of the small intestine by the northern blotting technique. Results: Hemoglobin and hepatic iron levels were statistically lower in the anemic rats. The mean transepithelial transports of sodium (−33.0 μEq · min−1 · cm−1), glucose (426.0 μM · min−1 · cm−1), and water (0.4 μL · min−1 · cm−1) in the small intestine of the anemic rats were significantly lower than in the control group (349.1 μEq · min−1 · cm−1, 842.6 μM · min−1 · cm−1, and 4.3 μl · min−1 · cm−1, respectively, P < 0.05). The transepithelial transport of potassium was similar for both groups. The relative SGLT1 mRNA levels of the anemic rats in the intermediate (1.796 ± 0.659 AU) and distal (1.901 ± 0.766 AU) segments were significantly higher than the values for the control rats (intermediate 1.262 ± 0.450 AU, distal 1.244 ± 0.407 AU). No significant difference was observed for the relative SLGT1 mRNA levels in the proximal segment or for the GLUT2 mRNA levels in all segments. Conclusion: Iron deficiency decreases the absorption of glucose, sodium, and water and increases SGLT1 mRNA in the intermediate and distal segments of the small intestine of rats. [Copyright &y& Elsevier]

Abstract: Intestinal glucose and calcium (Ca2+) absorption as well as their respective regulatory processes have been studied in detail in different species. Regulation is mediated from the luminal and the blood side of the barrier. For increasing glucose uptake, L-type Ca2+ channels in connection with cytoskeletal rearrangement have been identified as relevant factors for the insertion of glucose transporter 2 (GLUT2) into the apical membrane. Apical located sweet taste receptors can be stimulated by high luminal glucose concentrations or artificial sweeteners which also results in stronger GLUT2 insertion. During long-term adaptation, sweet taste receptors mediate the upregulation of a sodium-dependent glucose transport system (SGLT1), which involves the activation of cAMP pathway. Therefore, it is assumed that sweet taste receptors are able to modulate short-term as well as long-term adaptation through the activation of different pathways. For endocrine regulation glucagon-like peptides, glucose-dependent insulinotropic polypeptide and leptin are under discussion. For Ca2+ para- and transcellular pathways contribute to Ca2+ absorption which includes transient receptor potential channels (TRP). Controversy still exists on the essential role of calbindin-D9k for cellular transfer of Ca2+. Calcitriol, Ca2+ itself and other nutritional factors such as carbohydrates and short-chain fatty acids are involved in regulation of intestinal Ca2+ absorption. [ABSTRACT FROM AUTHOR]

Endurance exercise training can increase intestinal permeability which may contribute to the development of gastrointestinal symptoms in some athletes. Bovine colostrum (BC) supplementation reduces intestinal permeability induced by non-steroidal anti-inflammatory drugs. This study aimed to determine whether BC could also reduce intestinal permeability induced by endurance exercise. Thirty healthy adult males (25.0 ± 4.7 yr; mean ± SD) completed eight weeks of running three times per week for 45 minutes at their lactate threshold while consuming 60 g/day of BC, whey protein (WP) or control (CON). Intestinal permeability was assessed at baseline and after eight weeks by measuring the ratio of urinary lactulose (L) and rhamnose (R) excretion. After eight weeks the L/R ratio increased significantly more in volunteers consuming BC (251 ± 140%) compared with WP (21 ± 35%, P < 0.05) and CON (-7 ± 13%, P < 0.02). The increase in intestinal permeability with BC may have been due to BC inducing greater leakiness of tight junctions between enterocytes or by increasing macromolecular transport as it does in neonatal gut. Further research should investigate the potential for BC to increase intestinal macromolecular transport in adults. [ABSTRACT FROM AUTHOR]

Abstract: The effect of spironolactone (SL) pretreatment (200μmol/kg b.w./day, 3 consecutive days) on intestinal multidrug resistance-associated protein 2 (Mrp2) was evaluated in rats. A significant increase in protein levels in upper regions of small intestine, where Mrp2 is mainly present, was detected by western blotting. Real time PCR studies suggest a transcriptional regulation. The administration of ketoconazole, a pregnane X receptor (PXR) antagonist, was able to prevent the increase in Mrp2 mRNA levels induced by SL. The serosal to mucosal transport of dinitrophenyl S-glutathione, a model substrate of Mrp2 was evaluated in jejunal sac model. The data indicate that SL increased Mrp2 activity, well correlating with its up-regulation. We conclude that SL is able to induce intestinal Mrp2 transcriptionally, PXR being a potential mediator. We propose that SL could be of potential therapeutic application particularly in situations of down-regulation of intestinal Mrp2. [Copyright &y& Elsevier]

Abstract: The aim of the present study was to develop proliposomal formulations to enhance the oral bioavailability of exemestane by improving solubility, dissolution and/or intestinal permeability. Proliposomal powder formulations were prepared using different ratios of drug (exemestane), distearoyl–phosphatidylcholine (DSPC), cholesterol and dimyristoyl–phosphatidylglycerol (DMPG) by solvent evaporation method. The effect of phospholipid composition and drug:lipid ratio on in vitro performance of proliposomes was studied. Proliposomes were characterized for their particle size distribution, thermal characteristics by differential scanning calorimetry (DSC) and dissolution behavior. Further, the formulated proliposomes were subjected to in vitro permeation or transport studies using different models such as rat intestine, parallel artificial membrane permeability assay (PAMPA) and Caco-2 cell line. Proliposomes provided enhanced exemestane dissolution due to incorporation into the phospholipid bilayers and change in the physical state from crystalline to amorphous. The in vitro transport studies in rat intestine, PAMPA and Caco-2 models revealed that the proliposomes were successful in enhancing the permeation of exemestane. These proliposomal formulations of exemestane could provide improved oral bioavailability due to enhanced solubility, permeability and hence absorption. [Copyright &y& Elsevier]

Abstract: The use of solubilizing agents to improve the solubility of poorly water-soluble drugs often results in an alteration of intestinal membrane barrier function and intestinal membrane damage. In this study, 5(6)-carboxyfluorescein (CF) and fluorescein isothiocyanate-labeled dextran (MW 4400, FD4) were used as model compounds to examine the effects of twelve common solubilizing agents, sodium taurocholate (NaTC), Labrasol, polyethylene glycol 400 (PEG 400), Transcutol P, propylene glycol, Gelucire 44/14, HCO-60, ethanol, Cremophor EL, Tween 80, 2 hydroxypropyl-β-cyclodextrin (2HP-β-CyD) and dimethylsulfoxide (DMSO), on intestinal membrane barrier function and membrane toxicity in rats. Intestinal transport and absorption of CF were examined using an in vitro diffusion chamber and an in situ closed-loop technique. The in vitro diffusion chamber study showed that only 5 and 10% (w/v) NaTC significantly increased the transport of CF across the intestinal membrane. The in situ closed-loop study showed a remarkable increase in the absorption of CF and a bioavailability of more than 30% in the presence of 5 and 10% (v/v) Labrasol, 5 and 10% (w/v) NaTC and 10% (v/v) Transcutol P. Furthermore, we evaluated the effect of NaTC and Labrasol on the intestinal absorption of FD4, a high molecular weight compound. The results indicated that the absorption of FD4 also increased in the presence of 5 and 10% (w/v) NaTC and 10% (v/v) Labrasol, suggesting that these concentrations of NaTC and Labrasol may alter the intestinal membrane barrier functions in rats. We measured the release of protein and lactate dehydrogenase (LDH) from the intestinal membrane to examine the safety of solubilizing agents in the intestine. 5 and 10% (w/v) NaTC and 5 and 10% (v/v) Gelucire 44/14 significantly increased the presence of these toxicity markers compared to the control. The LDH level was also increased in the presence of 10% (v/v) of Cremophor EL. These findings suggest that the solubilizing agents at these concentrations except for NaTC, Gelucire 44/14 and Cremophor EL are considered safe and do not cause intestinal membrane damage. In conclusion, this study provides a basic approach in screening and predicting the effects of solubilizing agents for intestinal absorption studies using drugs poorly soluble in water. [Copyright &y& Elsevier]

Non-nutrient-dependent salt absorption across the brush-border membrane of intestinal epithelial cells is primarily mediated by coupled apical Na+/H+ (aNHE) and anion exchange transport, with the latter suspected of being mediated by DRA (downregulated in adenoma; SLC26A3) that is defective in congenital chloridorrhea. To investigate DRA in greater detail and determine whether DRA and NHE activities can be coupled, we measured 22Na+ and 36Cl- uptake in Caco2BBE colon cells infected with the tet-off-inducible DRA transgene. Under basal conditions, DRA activity was low in normal and infected Caco2BBE cells in the presence of tetracycline, whereas NHE activities could be easily detected. When apical NHE activity was increased by transfection or serum-induced expression of the aNHE isoforms NHE2 and NHE3, increased 36C1 uptake was observed. Inhibition of DRA activity by niflumic acid was greater than that by DIDS as well as by the NHE inhibitor dimethylamiloride and the carbonic anhydrase inhibitor methazolamide. DRA activity was largely aNHE-dependent, whereas a component of DRA-independent aNHE uptake continued to be observed. Coupled aNHE and DRA activities were inhibited by increased cellular cAMP and calcium and were associated with synaptotagmin I-dependent, clathrin-mediated endocytosis. In summary, these data support the role of DRA in electroneutral NaC1 absorption involving functional coupling of Cl-/base exchange and apical NHE. [ABSTRACT FROM AUTHOR]

Folate is a water-soluble B vitamin with a crucial role in the synthesis and methylation of DNA and in the metabolism of several amino acids. In the present study we investigated whether beverages like wine, beer and tea, or some of their specific constituents, affect the intestinal uptake of 3H-folic acid or 3H-methotrexate (an antifolate). All tested beverages significantly inhibited the uptake of 3H-folic acid by Caco-2 cells. Most of these beverages, with the exception of wines (not tested), also inhibited 3H-methotrexate uptake in these cells. Additionally, ethanol, when tested separately, inhibited the uptake of both compounds. Some of the tested phenolic compounds, namely myricetin, epigallocatechin gallate (EGCG) and isoxanthohumol, markedly inhibited 3H-folic acid uptake. Myricetin and EGCG also had a concentration-dependent inhibitory effect upon the uptake of 3H-methotrexate by Caco-2 cells. Resveratrol, quercetin and kaempferol were able to inhibit the transport of both compounds, but only in the concentration of 100 µM. In conclusion, dietary constituents may impact on intestinal folate uptake, as here shown for phenolic compounds. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) decreases glucose absorption in the proximal jejunum of laying hens in vitro and this effect is apparently mediated by the inhibition of the sodiumd-glucose co-transporter. DON could modulate the sugar transport of other intestinal regions of chickens. For this purpose, we have measured the effects of DON on the Na+d-glucose co-transporter, by addition of DON after and before a glucose addition in the isolated epithelium from chicken duodenum, jejunum, ileum, caecum and colon by using the Ussing chamber technique in the voltage clamp technique. The data showed in all segments of the gut that the addition ofd-glucose on the mucosal side produced an increase in the current (Isc) compared with the basal values, the Isc after glucose addition to the small intestine was greater than the Isc of the large intestine compared with the basal values, specially of the jejunum (p < 0.002), indicating that the jejunum is the segment that is the best prepared for Na+-d-glucose co-transport. Further addition of 10 μg DON/ml to the mucosal solution decreased the Isc in all segments and the Isc returned to the basal value, especially in the duodenum and mid jejunum (p < 0.05). In contrast, the addition of 5 mmold-glucose/l on the mucosal side after incubation of the tissues with DON in all segments had no effect on the Isc (p > 0.05), suggesting that DON previously inhibited the Na+d-glucose co-transport. The blocking effects of DON in duodenum and jejunum were greater than the other regions of the gut. It can be concluded that the small intestine of laying hens has the most relevant role in the carrier mediated glucose transport and the large intestine, having non-significant capacity to transport sugars, appears to offer a minor contribution to glucose transport because the surface area is small. The effect ofd-glucose on the Isc was reversed by DON in all segments, especially in the duodenum and jejunum, suggesting that DON entirely inhibited Na+-d-glucose co-transport. This finding indicates that the inhibition of Na+ co-transport system in all segments could be an important mode of action for DON toxicity of hens. [ABSTRACT FROM AUTHOR]

Abstract: In recent years, intestinal transport processes have been studied in detail regarding both, functional and structural aspects. For monosaccharides different systems have been demonstrated for apical uptake: this includes the high-affinity SGLT1 as a distinct d-glucose system and GLUT5 for fructose. Specifically in pigs a low affinity, high-capacity system for d-glucose and d-mannose with no preference for Na+ over K+ and a very low affinity system are suggested as further uptake systems. As in other species, basolateral extrusion is mediated by GLUT2. The distributions of monosaccharide transport along the gastrointestinal axis as well as the potential role of paracellular monosaccharide absorption have not yet been clarified. Amino acids can principally be absorbed by the paracellular and transcellular pathway whereas transcellular transport can either be mediated by facilitated diffusion or secondary active Na+-coupled transport. This includes different transport systems for neutral, anionic and cationic acids. In addition, the presence of the di-/tripeptides transport system PEPT1 which depends on an inwardly directed H+-gradient has also been confirmed for the pig small intestine, its quantitative proportion is still under debate. Short chain fatty acids (SCFA) are the major end products of microbial carbohydrate fermentation which occurs along the gastrointestinal tract with the highest production rates in the large intestines. At least two uptake mechanisms have to be assumed, i.e., non-ionic diffusion and anionic exchange via SCFA−/HCO3 −-exchange. Controversial views still exist to what extent SCFA are metabolized within the epithelial tissue. Segmental differences between small and large intestines have been demonstrated for Na+ absorption. Whereas in the small intestines the major part of Na+ absorption is mediated by coupled nutrient transport systems, aldosterone sensitive Na+ channels and Na+/H+-exchange are the dominant mechanisms in the hindgut. For Cl− paracellular transport and anionic Cl−/HCO3 −-exchange are the major absorptive mechanisms. Cl− secretion is mediated by apical channels which may be activated by toxins of different origin. Different types of Cl− channels have been identified, such as Cystic Fibrosis Transmembrane Regulator (CFTR), Ca-activated Cl− channels (CLCA) and Outwardly Rectifying Cl− Channels (ORCC). Whereas CFTR has clearly been shown for jejunal and colonic epithelial and goblet cells controversy still exists on the relevance of CLCA and ORCC in pigs. For Ca2+ there is evidence that both recently published channels TRPV5 and TRPV6 are also expressed in pig intestinal tissues, however, this has not yet been shown on protein level. From several functional approaches it was demonstrated that phosphate uptake can be mediated by both, a Na+-dependent transcellular component and paracellularly. On a molecular basis it is uncertain whether the transport protein of transcellular mechanism belongs to the NaPi-IIb cotransporter family. [Copyright &y& Elsevier]

Abstract: There has been an intensive debate whether dietary exposure to acrylamide could increase the risk of human cancer since the first description of the presence of acrylamide in food in 2002. As the intestinal mechanisms of acrylamide absorption are poorly investigated we studied the transport of acrylamide in differentiated Caco-2 cells and its effects on biotransformation enzymes (CYP2E1 and glutathione S-transferase) and glutathione levels. We found that the apparent permeability of [1-14C] acrylamide from the basal to the apical compartment was approximately 20% higher compared to that in the opposite direction. No differences were detected for apical–basal transport against a basal gradient. Transport rates from the apical to the basal chamber at 4°C were about 50% lower than at 37°C. Concentration dependent transport from apical to basal was linear. Predominantly, basal to apical transport was decreased when energy metabolism of the cells was inhibited by application of sodium azide and 2-deoxy-d-glucose. Finally, more acrylamide was transported at luminal pH 6 compared to pH 7.4 from basal to the apical direction. Increasing levels of acrylamide showed no effects on the activity of glutathione S-transferase but resulted in a depletion of total glutathione concentrations. In conclusion transport of acrylamide in the intestine is mediated primarily by passive processes possibly combined with a modest energy- and pH-dependent active secretory component. Depletion of cellular glutathione levels may be one potential mechanism for acrylamide (geno)toxicity. [Copyright &y& Elsevier]

ABCG5 and ABCG8 are two ATP-binding cassette half-transporters that belong to the G family members. They were identified as proteins that are mutated in a rare human disorder, sitosterolemia, and their identification led to the completion of the physiological pathways by which dietary cholesterol, as well as noncholesterol sterols, traffics in the mammalian body. These proteins are likely to function as heterodimers, and current evidence suggests that these proteins are responsible for the majority of sterol secretions into bile, thus may define the long sought-after biliary sterol transporters. This review will focus on some of the backgrounds of this physiology, the genetics and regulation of these genes, as well as our current understanding of their functions. This review will also highlight the current limitations in our knowledge gap. [ABSTRACT FROM AUTHOR]

It is generally accepted that 5-aminosalicylate (5-ASA; mesalamine), widely used in inflammatory bowel disease therapy, exerts its action from the intraluminal site of the intestine. In addition to local metabolism of 5-ASA, it has been assumed that therapeutic mucosal concentrations of 5-ASA depend on transporter-mediated secretion back to the lumen. We tested the hypothesis that 5-ASA represents a substrate of P-glycoprotein (P-gp) and/or multidrug resistance-associated protein 2 (MRP2), thereby possibly contributing to variable therapeutic effects. Polarized, basal-to-apical transport of [3H]5-ASA was studied in monolayers of Caco-2 and L-MDR cells, both of which express P-gp in their apical membrane, as well as in MDCK cells transfected with human MRP2. Moreover, we investigated the influence of 5-ASA on transport of digoxin in Caco-2 cells. In Caco-2 cells a P-gp-mediated efflux of 5-ASA (5–500 μM) could be excluded. Likewise, in L-MDR1 and MRP2 cells no transport differences in either the basal-to-apical or apical-to-basal direction were measurable. 5-ASA (50 μM to 5 mM) had no effect on the transport of digoxin. From these in-vitro experiments one can conclude that intestinal secretion of 5-ASA is apparently not mediated by P-gp or MRP2. Further studies are needed to identify the nature of the involved active carrier system(s). [ABSTRACT FROM AUTHOR]

Background: There is growing clinical interest in thalidomide for the treatment of various disorders due to its anti-inflammatory, immunomodulatory, and anti-angiogenic properties. In numerous clinical trials thalidomide is used as an adjunct to standard therapy. Therefore, clinicians should be aware of all possible drug–drug interactions that might occur with this drug. P-glycoprotein (P-gp), a drug efflux transporter that is expressed in many tissues, is the cause of several drug–drug interactions. P-gp induction or inhibition can lead to ineffective therapy or side-effects. In this study, we investigated thalidomide’s potential to cause drug–drug interactions on the level of P-gp. Methods: LS180 cells were incubated with thalidomide for 72 h in order to determine P-gp induction using real-time RT-PCR. A human leukaemia cell line over-expressing MDR1 (CCRF-CEM/MDR1) was used to measure uptake of rhodamine 123, a P-gp substrate, in the presence of thalidomide. Dose-dependent and bi-directional transport of thalidomide through Caco-2 cell monolayers was performed to assess site-directed permeability. Transport rates were determined using HPLC including chiral separation of the thalidomide enantiomers. Results: Thalidomide did not induce P-gp expression in LS180 cells. The uptake of rhodamine 123 in CCRF cells over-expressing MDR1 was not influenced by co-incubation with thalidomide. The transport through Caco-2 monolayers was linear and the permeability was similar for both directions. No differences between the thalidomide enantiomers were observed. Conclusions: Our study indicates that thalidomide is neither a substrate, nor an inhibitor or an inducer of P-gp. Therefore, P-gp-related drug–drug interactions with thalidomide are not likely. [ABSTRACT FROM AUTHOR]

The entry of most xeno/endobiotics into the organism is limited by their intestinal absorption. The interference of certain foods with the therapeutic efficacy of drugs or with chemical toxicity is becoming evident and growing attention is being given to these subjects. The aim of this work was to study the effect of green tea (GT) and black tea (BT), as well as some of their components, on the transport of organic cation molecules. For this purpose, 3H-MPP+ (radiolabeled 1-methyl-4-phenylpyridinium) was used as a model organic cation and Caco-2 cells were used as an intestinal epithelial model. Our results showed that both GT and BT significantly increased 3H-MPP+ absorption in these cells. Additionally, we studied the effect of epigallocatechin-3-gallate (EGCG), myricetin, caffeine, and theophylline. Whereas EGCG (2 mM) increased, myricetin (50 μM) and caffeine (1 mM) decreased, and theophylline (1 mM) had no effect on the uptake of 3H-MPP+ into Caco-2 cells. When GT was supplemented with caffeine or theophylline, we observed a partial loss of its effect. When BT was supplemented with EGCG, its ability to increase 3H-MPP+ uptake was much more pronounced than that observed with BT alone. In conclusion, this study showed that GT and BT might interfere with the absorption of the model organic cation MPP+ by the intestinal epithelium. Since important compounds are organic cations, the consequences of this interference may have an impact on human health. Although this constitutes only preliminary work and further studies are needed, tea should be included in the growing list of foodstuffs that have the potential to be involved in food–drug interactions. [ABSTRACT FROM AUTHOR]

The mechanism of uptake of dietary niacin (nicotinic acid) by intestinal epithelial cells is not well understood, and nothing is known about regulation of the uptake process. In this investigation, we used human-derived intestinal epithelial Caco-2 cells and purified intestinal brush-border membrane vesicles (BBMVs) isolated from human organ donors to assess niacin uptake. Our findings show niacin uptake by Caco-2 cells to be 1) temperature and energy dependent; 2) Na+ independent, but highly dependent on extracellular acidic pH; 3) saturable as a function of concentration, with an apparent Km of 0.53 ± 0.08 μM; 4) severely inhibited by the membrane-impermeable sulfhydryl group of reagents; and 5) highly specific for niacin but not affected by monocarboxylic acids. A marked trans stimulation in [3H]niacin efflux from preloaded Caco-2 cells by unlabeled niacin in the incubation buffer was also observed. These findings suggest the involvement of a specialized, pH-dependent, carrier-mediated mechanism for human intestinal niacin uptake. This suggestion was confirmed in studies with native human intestinal BBMVs. We also examined possible regulation of niacin uptake by Caco-2 cells via specific intracellular regulatory pathways. The results show that while the PKA-, PKC-, and Ca2+/ calmodulin-mediated regulatory pathways play no role in regulating niacin uptake, a role for a protein tyrosine kinase (PTK)-mediated pathway is apparent. The results of these studies show for the first time the existence of a specialized, acidic pH-dependent, carrier-mediated system of niacin uptake by human intestinal epithelial cells that operates at the micromolar (physiological) range of niacin. The results also suggest the possible involvement of a PTK-mediated pathway in the regulation of niacin uptake. [ABSTRACT FROM AUTHOR]

Background Milk products are a potential matrix for fortification with synthetic folic acid or natural 5-methyltetrahy- drofolate (5-CH3-H4folate) to enhance the daily folate intake. In milk, folate occurs bound to folate- binding proteins (FBP). Our previous studies with an in vitro gastrointestinal model showed that 70% of the initial FBP content of the milk product was retained in the duodenal lumen. While folic acid remained bound to FBP after gastric passage, 5-CH3-H4folate was mainly present as free folate in the duodenal lumen. Aim of the study To investigate the effect of FBP on the absorption of folic acid and 5- CH3-H4folate from the intestinal lumen. Methods The transport of [³H]-folic acid and [14C]-5-CH3- H4folate across enterocytes was studied in the presence or absence of bovine FBP using monolayers of Caco-2 cells grown on semi-permeable inserts in a two-compartment model. The apparent permeability coefficients (Papp) of folic acid and 5-CH3-H4folate were determined and compared with the permeability of reference compounds for low (mannitol) and high (caffeine) permeability. Results The transport from the apical to the basolateral side of the Caco-2 cells was higher (P< 0.05) for folic acid (Papp 1.7*10-6cm/s) than for 5-CH3-H4folate (Papp= 1.4*10-6cm/s) after 2 h incubation to 1 μM folic acid or 5-CH3-H4folate test solutions (pH 7). The permeability of folic acid and 5-CH3-H4folate across Caco-2 monolayers appeared to be higher (P < 0.05) than that of mannitol (Papp 0.5*10-6 cm/s) but lower (P < 0.05) than that of caffeine (Papp = 34*10-6 cm/s). The addition of FBP to the medium led to a lower (P<0.05) intestinal transport and cellular accumulation of folic acid and 5-CH3-H4folate. Conclusions Compared to the reference compounds, folic acid and 5-CH3-H4folate showed a moderate permeability across Caco-2 cells, which indicates that folate absorption from the intestinal lumen is not likely to be complete. The intestinal transport of folic acid and 5-CH3-H4folate was found to be dependent on the extent of binding to FBP at the luminal side of the cells. [ABSTRACT FROM AUTHOR]

The mechanisms involved in D-glucose and amino acid transport in the intestine of birds are still not clear. In chickens, D-glucose and amino acid absorption occurs via carrier-mediated transport, but in wild birds a passive paracellular mechanism seems to be the predominant pathway. The purpose of this work was to determine the existence of carrier-mediated sodium cotransport of D-glucose and L-alanrne in the small intestine of Japanese quail (Coturnix coturnix), a granivorous bird. Intestinal transport was determined by changes in the short-circuit current (Isc), proportional to ion trans-membrane flux, in the middle segment of the intestine of Japanese quail with a Ussing chamber. D-Glucose produced an increase of the Isc, and this effect was reverted by phloridzin, indicating the presence of a D-glucose transport mediated by the sodium/glucose cotranspoter 1. Addition of L-alanine also produced an increase of the Isc. We concluded that there is carrier-mediated cotransport of D-glucose and L-alanine with sodium in the small intestine of the Japanese quail. [ABSTRACT FROM AUTHOR]

Cystic fibrosis (CF) is frequently associated with progressive loss of exocrine pancreas function, leading to incomplete digestion and absorption of dietary fat. Supplementing patients with pancreatic lipase reduces fat excretion, but it does not completely correct fat malabsorption, indicating that additional pathological processes affect lipolysis and/or uptake of lipolytic products. To delineate the role of such (post) lipolytic processes in CF-related fat malabsorption, we assessed fat absorption, lipolysis, and fatty acid uptake in two murine CF models by measuring fecal fat excretion and uptake of oleate- and triolein-derived lipid. Pancreatic and biliary function was investigated by determining lipase secretion and biliary bile salt (BS) secretion, respectively. A marked increase in fecal fat excretion was observed in cftr null mice but not in homozygous ΔF508 mice. Fecal BS loss was enhanced in both CF models, but biliary BS secretion rates were similar. Uptake of free fatty acid was delayed in both CF models, but only in null mice was a specific reduction in lipolytic activity apparent, characterized by Strongly reduced triglyceride absorption. Impaired lipolysis was not due to reduced pancreatic lipase secretion. Suppression of gastric acid secretion partially restored lipolytic activity and lipid uptake, indicating that incomplete neutralization of gastric acid impedes fat absorption. We conclude that fat malabsorption in cftr null mice is caused by impairment of lipolysis, which may result from aberrant duodenal pH regulation. [ABSTRACT FROM AUTHOR]

The age-related changes of glutathione (GSH) levels and the effect of hypocaloric regimen and N-acetylcysteine (NAC) supplementation were investigated in intestinal mucosa and liver mitochondria of 28 months rats.Old rats exhibited lower proteins, GSH and protein sulphydrils (PSH) concentrations, higher GSH-peroxidase (GSH-Px) activity and protein carbonyl deposit, partial inhibition of succinate stimulated mitochondrial state III respiration and decreased mitochondrial nitrosothiols (RSNO) concentration. Lower electric potential and current intensity were found in the colonic mucosa.Old rats undergone hypocaloric regimen showed higher intestinal concentrations of GSH, lower oxidized protein accumulation and GSH-Px activity and higher mitochondrial RSNO levels. Mitochondrial state III respiration and intestinal transport were improved.NAC supplementation enhanced GSH and PSH levels in the ileal but not in the colonic mucosa, GSH and RSNO in liver mitochondria, while GSH-Px and protein carbonyls were decreased everywhere. Mitochondrial respiration ameliorated.In conclusion, ageing is characterized by a spread decrease of GSH concentrations, increased protein oxidation and decreased mitochondrial NO content. Hypocaloric diet ameliorated intestinal transport and, as well as NAC, was effective in enhancing GSH levels but at different extent according to the investigated districts. Both interventions reduced the age-associated increase of GSH-Px and protein carbonyls and improved mitochondrial respiration. [Copyright &y& Elsevier]

Hypertension or high blood pressure is a significant health problem worldwide. Bioactive peptides that inhibit angiotensin I converting enzyme (ACE) in the cardiovascular system can contribute to the prevention and treatment of hypertension. These ACE inhibitory peptides are derived from many food proteins, especially milk proteins. An ACE inhibitory activity in vitro does not always imply an antihypertensive effect in vivo. Even if it does, it is very difficult to establish a direct relationship between in vitro and in vivo activity. This is mainly due to the bioavailability of the ACE inhibitory peptides after oral administration and the fact that peptides may influence blood pressure by mechanisms other than ACE inhibition. To exert an antihypertensive effect after oral ingestion, ACE inhibitory peptides have to reach the cardiovascular system in an active form. Therefore, they need to remain active during digestion by human proteases and be transported through the intestinal wall into the blood. The bioavailability of some ACE inhibitory peptides has been studied. It is also known that (hydroxy)proline-containing peptides are generally resistant to degradation by digestive enzymes. Peptides can be absorbed intact through the intestine by paracellular and transcellular routes, but the potency of the bioactivity after absorption is inversely correlated to chain length. In addition, some strategies are proposed to increase the bioavailability of ACE inhibitory peptides. Further research into the bioavailability of ACE inhibitory peptides will lead to the development of more effective ACE inhibitory peptides and foods. [ABSTRACT FROM PUBLISHER]

The intestinal absorptive characteristics and the efflux mechanisms of 9-nitrocamptothecin (9-NC), a novel water-insoluble camptothecin (CPT) derivative, were investigated. The Caco-2 cells and the everted gut sacs were used as models of the intestinal mucosa to assess transepithelial transport of 9-NC. The determination of 9-NC was performed by HPLC. In the Caco-2 cells, the absorptive transport of 9-NC was pH dependent and the transport was enhanced at weakly acidic pH on the apical side. No concentration dependence and saturation were observed for the absorptive transport of 9-NC at concentrations up to 250 μM, while secretory transport were concentration dependent and saturable process (Km was 49.8±1.2 μM, Vmax was 38.28±0.8 ng/cm2/min). In the presence of verapamil (100 μM) and CsA (10 μM), potent inhibitors of P-glyprotein (P-gp)/MRP2 (cMOAT), the Papp BL-AP/Papp AP-BL ratio was decreased from 3.4 to 1.4 and 1.3, respectively, and permeation of apical to basolateral was enhanced approximately two-fold. In the everted gut sacs, the absorption of 9-NC was passive diffusion and had no significant difference in different gut regions. Adding verapamil in the everted gut sacs over a concentration ranging from 10 to 100 μM, the absorption of 9-NC was significantly enhanced, especially more markedly in lower small intestine (P<0.05). Overall, the current study suggests that pH and efflux transporters are capable of mediating the absorption and efflux of 9-NC, and they may play significant roles in limiting the oral absorption of 9-NC. [Copyright &y& Elsevier]

Background: Approximately 25% of women retain 5 kg of the weight gained during pregnancy, but the physiologic factors underlying excessive postpartum weight gain are not known. Objective: The objective of the study was to determine whether pregnancy-related adaptive increases in intestinal nutrient transport are retained after parturition and therefore contribute to postpartum weight gain. Design: We measured body weight and intestinal nutrient transport in virgin (V, control), primiparous (P, one pregnancy), and multiparous (M, 3 pregnancies) mice at parturition (day 1), during lactation (days 14 and 21), at weaning (day 28), after weaning (day 40), and during aging (days 70, 120, 200, and 300). Results: In M and P mice, body weight and the weight and length of the small intestine were greatest during lactation; they then decreased but did not return to prepregnancy values until 300 d after parturition. Intestinal villus heights were maximal at lactation and remained high ⩽ 200 d after parturition. Total intestinal transport capacity for D-glucose, D-fructose, and L-proline was also greatest during lactation, and the lactation-enhanced transport capacity was retained ⩽ 70 d after parturition. M mice retained more body weight and intestinal transport capacity postpartum than did P mice. Aging per se had little or no effect on body weight or intestinal weight, length, and nutrient transport. The dramatic, lactation-related increases in intestinal nutrient transport capacity were due mainly to increases in intestinal mass. Conclusions: Postpartum retention of pregnancy- and lactationrelated increases in intestinal nutrient uptake capacity may play a significant role in postpartum body weight retention. These adaptive increases may be cumulative and may result in greater weight retention in mice with multiple pregnancies. [ABSTRACT FROM AUTHOR]

Caco-2 epithelial layers were used as a model to re-evaluate the mechanism(s) by which intestinal digoxin absorption is limited by its active secretion back into the lumen. It is widely recognised that intestinal secretion of digoxin is mediated by the ATP-binding cassette (ABC) transporter Multidrug Resistance 1, MDR1. In MDR1-transfected Madin-Darby canine kidney, MDCKII , cell monolayers, digoxin secretion was reduced by the MDR1 inhibitor cyclosporin A, whereas no inhibition was seen in the presence of MK-571, 3-([(3-(2-[7-chloro-2-quinolinyl]ethyl)phenyl]-[(3-dimethylamino-3-oxoprphyl)-thio)-methyl]-thio) propanoic acid, a Multidrug Related Protein (MRP) inhibitor. In contrast, digoxin secretion by Caco-2 epithelia was significantly inhibited by both cyclosporin A and MK-571, suggesting that an additional non-MDR1 component may contribute to this transport. Since digoxin secretion by MRP2-transfected MDCKII monolayers was increased by only 1.2-fold relative to controls, it is likely that the contribution of MRP2 to digoxin secretion by Caco-2 cells is negligible. An additional MK-571-sensitive secretory pathway for digoxin, together with MDR1, is likely to mediate digoxin secretion in Caco-2 epithelia. [Copyright &y& Elsevier]

The antidiabetic biguanide metformin has been shown to increase faecal excretion of bile salts in type 2 diabetes. Cultured human intestinal Caco-2 cell monolayers provide a model of human enterocytes. These monolayers are used here to determine the effect of metformin on the secondary-active, sodium-linked transfer of 14 C-glycocholate from the apical (brush border) to the basolateral (serosal) surface. During 24-h incubations, 10-2 mol/l metformin significantly reduced 14 C-glycocholate transfer. This could not be attributed to alterations of monolayer integrity or Na+ -K+ ATPase pump activity. For example, the secondary-active transport of glucose and proline was not interrupted, and the inhibitory effect of metformin on bile salt transport was additive to the inhibitory effect of ouabain. The results suggest that metformin can act directly on intestinal enterocytes to reduce the active transfer of bile salts by a mechanism that is independent of Na+ -K+ ATPase activity. [ABSTRACT FROM AUTHOR]

Human milk contains a variety of complex lactose derived oligosaccharides in relative high amounts which are only minimally digested during their passage through the orogastrointestinal tract [Gnoth, M. J., Kunz, C., Kinne-Saffran, E., & Rudloff, S., (2000a). Human milk oligosaccharides are minimally digested in vitro. Journal of Nutrition, 130, 3014–3020] Since these components were detected in the urine of breast-fed infants [Obermeier, S., Rudloff, S., Pohlentz, G., Lentze, M. J., & Kunz, C. (1999). Section of 13C-labelled oligosaccharides into human milk and infant''s urine after an oral [13C]galactose load. Isotopes Environmental Health Studies, 352, 119–125], we investigated possible intestinal transport mechanisms using confluent Caco-2 layers grown on permeable membrane filter supports. When the apical side of these cell layers was exposed to HMO virtually the complete milk oligosaccharide pattern could be detected on the basolateral side. For neutral, but not for acidic oligosaccharides, the transport from the apical to the basolateral compartment was higher than in the opposite direction. Quantitatively, the transport from the apical to the basolateral compartment was 1–1.5% of the applied amount of oligosaccharides. [Copyright &y& Elsevier]

Growth factor-stimulated intestinal absorption has recently been described, but the cellular transport mechanisms mediating this response are unknown. The purposes of this study were to examine the effect that intraluminal and systemic EGF and TGF have in intestinal absorption, elucidate a possible mechanism through which they exert their activity, and compare this response to that of a mixed meal only. Jejunal and ileal Thiry-Vella intestinal segments were constructed in six dogs. Absorption was measured by infusing the loops with a physiological electrolyte solution containing either 10 mmol or 50 mmol glucose and [14C]PEG as the impermeant marker. In vivo studies show that the addition of either EGF or TGF resulted in increased absorption of Na+, Cl-, H2O, and glucose in the intestine. This response was significantly greater than that seen when giving a mixed meal alone. Luminal phloridzin, an inhibitor of the SGLT-1 transporter, inhibited intestinal absorption observed in response to EGF and TGF. In conclusion, these results suggest that growth factors are capable of up-regulating intestinal absorption of electrolytes and nutrients and, these effects are mediated, at least in part, by SGLT-1 pathways. [ABSTRACT FROM AUTHOR]

Iron deficiency induced in rats maintained on a commercial diet with a low iron content has been used to investigate adaptive mechanisms that enhance duodenal iron uptake. These adaptive changes have been divided into those that result from changes in villus surface area (structural adaptation) and those that reflect changes in the way individual enterocytes express iron transport function (cellular adaptation). Cellular adaptation was assessed by carrying out microdensitometry of autoradiographs prepared from duodenal tissue previously incubated for 5 min in 200 µmol/l 59Fe2+-ascorbate. Structural adaptation was studied by performing image analysis of microdissected and sectioned villi. Cellular adaptation involved increased iron uptake by enterocytes present in the lower villus. Thus iron deficiency resulted in a threefold enhanced expression of uptake in the lower 100 µm villus (3.9±2.4 versus 12.6±1.5 arbitrary units, P<0.001). Maximal uptake was reached in the upper region of both control and iron-deficient villi, but iron deficiency had no effect on cellular uptake at this part of the villus. Structural adaptation involved the lengthening (+16%, P<0.05) and broadening (+14%) of villi in the duodenum of iron-deficient rats. The resultant expansion in villus area caused a further increase in uptake that was mostly expressed in the upper villus. Maximal uptake corrected for structure occurred in the middle third of villi from control and iron-deficient rats. Cellular plus structural adaptation produced a twofold increase in iron uptake. More than half of this effect was caused by changes in villus structure. [3H]Thymidine labelling experiments revealed a slightly earlier expression of enterocyte iron uptake in iron deficiency. [ABSTRACT FROM AUTHOR]

Zinc oxide (ZnO) nanoparticles (NPs) are used as zinc supplements due to the nutritional value of Zn. The toxicity of ZnO NPs in the food industry is required to be elucidated because they have large surface area and high reactivity compared with bulk-sized materials and have potentials to interact with food matrices, which may lead to different biological responses. In this study, interactions between ZnO NPs and food proteins (albumin, casein, and zein) were evaluated by measuring changes in physicochemical property, fluorescence quenching ratios, and structural protein stability compared with ZnO interaction with glucose, the most interacted saccharide in our previous report. The interaction effects on cytotoxicity, cellular uptake, intestinal transport, toxicokinetics, and acute oral toxicity were also investigated. The results demonstrate that interaction between ZnO and albumin reduced hydrodynamic diameters, but increased cytotoxicity, cellular uptake, and intestinal transport in a similar manner to ZnO interaction with glucose, without affecting primary structural protein stability and toxicokinetic behaviors. Hematological, serum biochemical, and histopathological analysis reveal no toxicological findings after orally administered ZnO NPs interacted with albumin or glucose in rats for 14 consecutive days, suggesting their low oral toxicity. In conclusion, the interactions between ZnO NPs and food proteins modulate in vitro biological responses, but do not affect in vivo acute oral toxicity. Further study is required to ascertain the interaction effects on chronic oral toxicity. [ABSTRACT FROM AUTHOR]

Purpose. To determine if intestinal secretion occurs for the poorly bioavailable diuretic, furosemide. Methods. Jejunal segments of male Sprague-Dawley rats were mounted on diffusion chambers, and the permeation of furosemide was measured across the excised tissue in both directions. Studies were repeated using cultured epithelia from adenocarcinoma cells (Caco-2) grown on filter inserts mounted in 6-well plates. Temperature-dependence and chemical inhibition by indomethacin was also tested using the cell culture model. Results. Net secretion from rat intestine of over 3-fold was observed for 20 µM furosemide. Net secretion of furosemide by Caco-2 cells was over 300% greater than for intestinal segments (10-fold vs. 3-fold). For both models, a decrease in furosemide transport in the direction of secretion was observed in the presence of indomethacin (100 µM), although only results using the Caco-2 cells showed an increase in the absorptive transport. Furosemide secretion from Caco-2 cells decreased with decrease in temperature from 37°C to 4°C, suggesting a carrier-mediated process. Conclusions. Furosemide appears to be secreted in the small intestine. These preliminary results indicate that furosemide bioavailability may be limited by an intestinal transporter. [ABSTRACT FROM AUTHOR]

The ordinary aliphatic, neutral amino acids and phenylalanine have been examined for cis-inhibition of influx of alanine ( J) and lysine ( J) and trans-stimulation of J across the brush border membrane of rat small intestines: and their effects on the unidirectional mucosa-to-serosa flux ( J) across the short circuited intestine have been studied. The effects of alanine, α-amino- n-butyric acid, leucine, and methionine on the steady-state epithelial uptake of lysine [Lys] have also been measured. In addition the trans-effects of alanine and leucine have been examined for sodium-dependence, and alanine was tested as trans-stimulator of influx of galactose across the brush border membrane ( J). All the neutral amino acids were found to be competitive cis-inhibitors of J, and all, except isoleucine, were trans-stimulators of J. The magnitude of the trans-effect was unrelated to the efficiency of the amino acid as cis-inhibitor. As illustrated by alanine, the trans-effects are probably completely sodium-dependent. Alanine was also effective as trans-stimulator of J. With respect to effects on [Lys] and J the neutral amino acids fall into two groups: One which reduces [Lys] and stimulates J, and one which increases [Lys] and relatively inhibits J. These effects are not correlated with the affinities of the neutral amino acids for the two carriers involved. It is proposed that the trans-effects on J are induced by an electrogenic, sodium-coupled efflux of the neutral amino acid across the brush border membrane, that the stimulation of J is brought about by a selective stimulation (of unknown nature) of efflux of lysine across the basolateral membrane ( J), assisted by competitive inhibition of lysine efflux across the brush border membrane ( J), and that the amino acids which do not stimulate J increase [Lys] by competitively inhibiting J and J. The inhibitory effect of the neutral amino acids on J support the view that the carrier of basic amino acids serves as a second carrier of these amino acids. [ABSTRACT FROM AUTHOR]

Without interruption of the blood flow, the injection of formalin (10% formaldehyde) for ten minutes into the lumen of isolated loops of dog ileum produces a corrosive destruction of the mucosa down to the level of the sub-mucosa. The regeneration of the epithelium from the borders of the intact intestine and the formation of new villi have been followed for four weeks post-operatively. One day after the treatment, a necrotic band shows the depth of the destruction, which varies from one dog to another. After one week, the loops are already shortened by 20-40% of their original length and the surface of the intestine is covered with granulation tissue. At first an unicellular layer of cells progresses from the untreated mucosa. In addition, in rare cases, islets of intact cells within the granulation tissue and in the middle of the loops can be detected. These are generally located in the vicinity of Peyer's patches. The first structures of newly-formed villi reappear three weeks postoperatively, when the functional capacity of the epithelial cells of the regenerated mucosae is only slightly below that of normal tissue. After four weeks, one third of the loops are shortened to about 40% of their original length; they are stenotic to varying degrees and without signs of epithelial covering. In these cases, the original injury had obviously reached the sub-mucosa. In the remaining two thirds of the loops where considerably less reduction was encountered, the regeneration and formation of villi has advanced, although small ulcerations can still be detected, and the function of the mucosa does not differ significantly from that of contiguous control loops. [ABSTRACT FROM AUTHOR]

The properties of hexose transport across the apical and basolateral membranes of chicken enterocytes have been studied in the small and large intestine. Results show that (a) isolated epithelial cells from all segments except the coprodeum can accumulate 3- O-methylglucose (Glc3Me) against a concentration gradient, by a Na-dependent and phloridzin-sensitive mechanism, (b) The cell cumulative capacity for Glc3Me (control/phloridzin-incubated cells) is lower in the small intestine than in the large intestine (rectum = proximal caecum = ileum > jejunum > duodenum). (c) Theophylline enhances the cell Glc3Me cumulative capacity 2.9-fold in the duodenum and 2.4-fold in the jejunum but has no effect in the other segments studied. (d) Analysis of sugar uptake indicates that net hexose influx rates decrease from proximal to distal regions: jejunum > duodenum > ileum = proximal caecum = rectum for the apical transport system ( α-methyl glucoside as substrate and phloridzin as inhibitor) and duodenum > jejunum > ileum = proximal caecum = rectum for the basolateral system (2-deoxyglucose; theophylline). (e) The duodenum and the jejunum show high apical and basolateral hexose transport rates, which confer a significant capacity for sugar absorption on the proximal intestine. More distal regions, including the ileum, the proximal caecum and the rectum, have transport systems analogous to those of the proximal intestine that keep a considerable potential capability to recover hexoses from the lumen. [ABSTRACT FROM AUTHOR]

The aims of the present study were: (1) to set up a procedure for simultaneous vascular and luminal perfusion of the chicken jejunum; (2) to assess the transport capacity of the tissue under such conditions, and (3) to study the effects of phloretin and theophylline, given through the vascular perfusate, on 3- O-methyl- D-glucose intestinal transport. The perfusion procedure described allowed the control of intestinal and vascular inflow rates and perfusion pressures so that these parameters could be adjusted to physiological values. A perfluorochemical emulsion was used as oxygen carrier for the vascular perfusate. The absorptive function of the perfused tissue was assessed by means of its ability to transport 3- O-methyl- D-glucose. Furthermore, ultrastructure preservation was evaluated by scanning and transmission electron microscopy. Results indicate that the perfused tissue kept its transport capacity and morphology intact throughout the 120-min experimental period. Moreover, no hypersecretion was observed as indicated by the constancy of perfusate volumes and perfusion pressures. Phloretin (1 mM) or theophylline (10 mM) added to the vascular perfusate markedly reduced the transfer of 3- O-methyl- D-glucose from the enterocyte to the vascular fluid without affecting the uptake from the lumen. Our results suggest that this preparation may be used as an alternative tool for the study of intestinal absorption processes in avian species, particularly when complete examination is required of the efflux of substrates from the intestinal lumen to the vascular fluid. [ABSTRACT FROM AUTHOR]

Intestinal epithelial cells isolated from rabbit small intestine and whose plasma membrane had been rendered highly permeable were used to study the role of intracellular structures in Ca buffering. Monitoring free Ca with a selective electrode revealed that the cells could reduce Ca concentration in the medium to a level of 3.6×10 M independently, within a certain range, of the initial Ca concentration or amount of cells used. Ca buffering by permeabilised enterocytes was Mg-and ATP-dependent and was abolished in the presence of the Ca ionophore A23187. The rapidity of Ca buffering, but not the final Ca level attained, was reduced in the combined presence of the mitochondrial inhibitors azide and oligomycin or in the presence of ruthenium red. Buffering of Ca was abolished in the presence of vanadate, although some uptake was still observed. Complete blocking occurred in the presence of hoth vanadate and mitochondrial inhibitors. Measurement of initial rates of uptake with radioactive calcium, revealed that mitochondrial uptake plays a role at relatively high Ca concentrations but that at the presumably physiological levels most of the uptake is into a nonmitochondrial compartment. Non-differentiated crypt cells scemed to handle intracellular Ca in a similar way as mafure villus cells, although they appeared to buffer at a level about 2×10 M lower. [ABSTRACT FROM AUTHOR]

The effects of forskolin on mucosal cyclic AMP levels and active transport of glycine, l-lysine and l-leucine were studied in rat jejunum in vitro. Furthermore, the effects on lysine and glycine incorporation into mucosal protein and on mucosal cell volume were investigated. Elevation of intestinal mucosal cyclic AMP to threefold control levels by 10 μmol l forskolin was accompanied by increased absorption of glycine (+33%), l-leucine (+72%) and l-lysine (+188%), as determined in a three compartment model suitable to measure active transport. Increased intracellular accumulation could be demonstrated for lysine as a transport substrate. Accordingly, using a dual label method, calculated values for uphill transport of lysine at the site of the brush border membrane were markedly enhanced. Forskolin up to 10 μmol l had no effects on the fraction of lysine or glycine incorporated into TCA-precipitable proteins of jejunal absorptive cells. Serosal to mucosal transfer, as well as basolateral entry into mucosal cells remained unchanged for all three amino acids. Likewise, intracellular fluid space, calculated from distribution spaces for C-inulin and HO as well as the response of cellular volume to an osmotic gradient were not affected by forskolin. As comparable stimulatory effects of forskolin on active hexose transport were reported earlier, it is suggested that forskolin-known to inhibit sodium-coupled fluid absorption-may stimulate active transport by enhancing sodium availability for sodium dependent intestinal cotransporters in general. [ABSTRACT FROM AUTHOR]

The phosphodiesterase-inhibitor 3-isobutyl-1-methylxanthine (IBMX) was able to elevate rat small intestinal cyclic AMP levels to 300% of basal values. Active jejunal d-glucose transport was enhanced parallel to the rise of intracellular cyclic AMP levels to 140% of control values at 100 μmol/l IBMX. Transport parameters, as determined in a three compartment model in vitro using a dual label method, indicate increased 'uphill' glucose transport at the site of the brush border membrane, higher intracellular accumulation of the sugar, with unchanged passive permeabilities. Phlorizin-inhibited d-glucose transport and l-glucose transfer in the rat were not affected by the persisting cyclic AMP elevation produced by IBMX. Stimulating effects could also be demonstrated with d-galactose as a substrate. IBMX 100 μmol/l also increased active d-glucose as well as 3-O-methylglucose transport in mouse jejunum. Stimulatory effects on intestinal hexose transport and mucosal cyclic AMP levels were also found with the adenylate-cyclase activator forskolin. In the present study, forskolin effects on jejunal mucosal cyclic AMP levels were enhanced in the presence of 100 μmol/l IBMX, resulting in a 20-fold increase compared to controls at 20 μmol/l forskolin. The concentration response for the effect of forskolin in the presence of 100 μmol/l IBMX on d-glucose transport did not produce a significant increase compared to transport stimulation with IBMX alone. At higher concentrations of forskolin however, glucose transport decreased to levels well below the IBMX controls. The elevation of cellular cyclic AMP levels had no effects on passive permeability. Both IBMX 100 μmol/l as well as forskolin 20 μmol/l inhibited rat jejunal net fluid transport by 40%, combination of both agents resulted in a 55% reduction of net fluid absorption in everted sacs of rat jejunum. These results indicate a functional relationship between jejunal mucosal cyclic AMP levels and active hexose absorption different from the inhibitory role of cyclic AMP in intestinal fluid transport. [ABSTRACT FROM AUTHOR]

The adenylate cyclase stimulator forskolin enhanced both mucosal cyclic AMP levels and d-aldohexose transport in rat jejunum in vitro in a concentration-dependent manner. With mucosal cyclic AMP elevated to 400% of basal values, active d-glucose transport was at 200% of control values. Transport parameters, as determined in a three compartment model using a dual label method, indicate a) increased 'uphill' glucose transport at the brush border membrane with higher intracellular accumulation, b) unchanged serosal to mucosal glucose fluxes and passive permeabilities. Transport stimulating effects of forskolin were also present with d-galactose in the rat. In mouse jejunum active transport of d-glucose and 3-O-methylglucose was increased by forskolin 10 μmol/l. Phlorizin inhibited d-glucose transport as well as l-glucose transport in the rat were not changed by the persisting cyclic AMP elevation induced by forskolin. The results show a positive correlation of active intestinal hexose transport with a modest elevation of mucosal cyclic AMP. In accordance to current models of cyclic AMP-induced changes in intestinal fluid absorption, rat jejunal net fluid transport was reduced by 40% with 20 μmol/l forskolin. [ABSTRACT FROM AUTHOR]