Glucose is the main energy source for mammalian cells and its absorption is co-mediated by two different families of glucose transporters, sodium/glucose co-transporters (SGLTs) and facilitative glucose transporters (GLUTs). Here, we report the cloning and tissue distribution of porcine GLUT2. The GLUT2 was cloned by RACE and its cDNA was 2,051 bp long (GenBank accession no. EF140874). An AAATAA consensus sequence at nucleotide positions 1936-1941 was located upstream of the poly $(A)^+$ tail. Open reading frame analysis suggested that porcine GLUT2 contained 524 amino acids, with molecular weight of 57 kDa. The amino acid sequence of porcine GLUT2 was 87% and 79.4% identical with human and mouse GLUT2, respectively. GLUT2 mRNA was detected at highest level in porcine liver, at moderate levels in the small intestine and kidney, and at low levels in the brain, lung, muscle and heart. In the small intestine, the highest level was in the jejunum. In conclusion, the mRNA expression of GLUT2 was not only differentially regulated by age, but also differentially distributed along the small intestine of piglets, which may be related to availability of different intestinal luminal substrate concentrations resulting from different food sources and digestibility.
A low level exposure experiment was conducted on growing rats to investigate the accumulation and organ distribution of protein bound cadmium compared with cadmium chloride. Male Sprague-Dawley rats were fed for 21days with one of the semisynthetic diets, which contains cadmium as either bovine liver- or kidney meal bound cadmium, cadmium chloride with uncontaminated liver meal or cadmium chloride without organ meal, in the levels of ca. 0.5, 1 and 1.5mg/kg diet, respectively. After 21days of exposure cadmium was accumulated in liver, kidney and gastrointestinal tracts depending upon cadmium levels in diet. Inspite of very low cadmium accumulation in whole blood, it tends also to increase with dietary cadmium levels. The blood cadmium concentration of animals fed organ meal containing diets was about 4-7 fold higher than that without organ meal, regardless of cadmium was intrinsically bound to protein or not. However, significant effects of organ protein on cadmium accumulation in liver, kidney and digestive tracts were not detectable, when cadmium was supplemented as cadmium chloride. On the other hands, animals fed diet containing ca. 1.5mg Cd/kg as organ bound cadmium retained more cadmium in liver, kidney and digestive tracts compared to cadmium chloride with organ meal, whereby the increase of cadmium concentration in kidney was greater then in liver. However, when the concentration of protein bound cadmium was<1mg/kg diet, organ bound cadmium was not significantly different from cadmium chloride in bioavailability and organ distribution. From this result it is suggested that the intestinal absorption of protein bound cadmium is influenced of the amount of cadmium bound in protein. When cadmium concentration in protein is relatively low, protein bound cadmium seems to be absorbed in the same way as cadmium ions are absorbed. However, when the concentration is high, at least a small amount of intact protein bound cadmium could be absorbed and accumulated selectively in kidney.
Growing pigs (N=25; 18 kg) were used to study effects of L-carnitine and protein intake on plasma carnitine, energy and carnitine balance, and carnitine biosynthesis. Corn-soybean meal basal diets containing low or high protein (13.6% or 18%) were formulated so that protein accretion would be limited by metabolizable energy (ME). Each basal diet was supplemented with 0 or 500 mg/kg L-carnitine and limit fed to pigs for 10 d in a balance trial. Final carnitine concentration was compared with weight/age matched pigs measured on d 0 to calculate carnitine retention rates. Supplementation of carnitine increased (p<0.01) plasma free carnitine (by 250%), short-chain (by 160%) and long-chain acyl-carnitine concentrations (by 80%) irrespective of blood sampling time (p<0.01). The proportion of long-chain carnitine esters decreased by 40% (p<0.01) by carnitine supplementation; whereas, the proportion of short-chain acyl-carnitine concentration was not changed (p>0.10). All criteria of energy balance were unaffected by L-carnitine (p>0.10). Total body carnitine retention was increased by 450% over unsupplemented controls (p<0.01). Carnitine biosynthesis rates in pigs fed diets without L-carnitine were estimated at 6.71 and $10.63{\mu}mol{\cdot}kg^{-1}{\cdot}d^{-1}$ in low protein and high protein groups, respectively. In supplemented pigs, L-carnitine absorption and degradation in the intestinal tract was estimated at 30-40% and 60-70% of L-carnitine intake, respectively. High protein feeding effect did not affected plasma carnitine concentrations, carnitine biosynthesis or carnitine retention (p>0.10). We conclude that endogenous carnitine biosynthesis may be adequate to maintain sufficient tissue levels during growth, but that supplemental dietary carnitine (at 500 ppm) sufficiently increased plasma acyl-carnitine and total body carnitine.
Lee, Eui Sup;Seo, Hyun Jung;BacK, Su Sun;Han, Seung Ho;Jeong, Yeon Ji;Lee, Jin Wook;Choi, Soo Young;Han, Kyuhyung
BMB Reports
/
제48권9호
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pp.513-518
/
2015
Factors that modulate cholesterol levels have major impacts on cardiovascular disease. Niemann-Pick C1-like 1 (NPC1L1) functions as a sterol transporter mediating intestinal cholesterol absorption and counter-balancing hepatobiliary cholesterol excretion. The liver receptor homolog 1 (LRH-1) had been shown to regulate genes involved in hepatic lipid metabolism and reverse cholesterol transport. To study whether human NPC1L1 gene is regulated transcriptionally by LRH-1, we have analyzed evolutionary conserved regions (ECRs) in HepG2 cells. One ECR was found to be responsive to the LRH-1. Through deletion studies, LRH-1 response element was identified and the binding of LRH-1 was demonstrated by EMSA and ChIP assays. When SREBP2, one of several transcription factors which had been shown to regulate NPC1L1 gene, was co-expressed with LRH-1, synergistic transcriptional activation resulted. In conclusion, we have identified LRH-1 response elements in NPC1L1 gene and propose that LRH-1 and SREBP may play important roles in regulating NPC1L1 gene. [BMB Reports 2015; 48(9): 513-518]
Although an adequete intake of calcium (Ca) is recommended for the treatment and prevention of osteoporosis, the intake of Ca should be restricted because of its low rate of intestinal absorption. The purpose of this experiment was to identity the effect of the combined administration of Aquamin F (AQF) (a calcium agent) and lactic acid bacteria (LAB) on osteoporosis. Thirty ovariectomized (OVX) rats and six control rats were assigned to the following six groups, with six animals per group: sham Ca-deficient diet (Ca-D), OVX, LAB, AQF, and LAB-AQF. During the experiment, the body weight was measured; and after the experiment was completed, the serum biochemical analysis, the alkaline phosphatase, calcium, and inorganic phosphorus leves were measured. The tissue of the femur was stained and then scanned via CT. The body weight of the OVX group increased more significantly than that of the control group. The results of the bone mineral content (BMC), Bone mineral density (BMD), serum biochemical analysis and histological test on the femur epiphysis showed no difference between the OVX group and the LAB group, whereas the results of the AQF group were more significant than those of the OVX group. In particular, the LAB+AQF group showed more significant increases in the aforementioned results than the AQF group. This experiment showed that the combined administration of AQF and LAB in ovariectomized rats more significantly increased bone density than did a single administration of either AQF or LAB.
This study aimed to investigate the in vivo relevance of P-glycoprotein (P-gp) in the pharmacokinetics and adverse effect of phenformin. To investigate the involvement of P-gp in the transport of phenformin, a bi-directional transport of phenformin was carried out in LLC-PK1 cells overexpressing P-gp, LLC-PK1-Pgp. Basal to apical transport of phenformin was 3.9-fold greater than apical to basal transport and became saturated with increasing phenformin concentration ($2-75{\mu}M$) in LLC-PK1-Pgp, suggesting the involvement of P-gp in phenformin transport. Intrinsic clearance mediated by P-gp was $1.9{\mu}L/min$ while passive diffusion clearance was $0.31{\mu}L/min$. Thus, P-gp contributed more to phenformin transport than passive diffusion. To investigate the contribution of P-gp on the pharmacokinetics and adverse effect of phenformin, the effects of verapamil, a P-gp inhibitor, on the pharmacokinetics of phenformin were also examined in rats. The plasma concentrations of phenformin were increased following oral administration of phenformin and intravenous verapamil infusion compared with those administerd phenformin alone. Pharmacokinetic parameters such as $C_{max}$ and AUC of phenformin increased and CL/F and Vss/F decreased as a consequence of verapamil treatment. These results suggested that P-gp blockade by verapamil may decrease the phenformin disposition and increase plasma phenformin concentrations. P-gp inhibition by verapamil treatment also increased plasma lactate concentration, which is a crucial adverse event of phenformin. In conclusion, P-gp may play an important role in phenformin transport process and, therefore, contribute to the modulation of pharmacokinetics of phenformin and onset of plasma lactate level.
5-Aminosalicylic acid (5-ASA) is an active ingredient of therapeutic agents used for Crohn s disease and ulcerative colitis. Because it is absorbed rapidly and extensively in the upper intestine, delivery of the agent specifically to the colon is necessary. We selected taurine as a colon-specific promoiety and designed 5-aminosalicyltaurine (5-ASA-Tau) as a new colon-specific prodrug of 5-aminosalicylic acid (5-ASA). It was expected that introduction of taurine would restrict the absorption of the prodrug and show additive effect to the anti-inflammatory action of 5-ASA after hydrolysis. 5-ASA-Tau was prepared in good yield by a simple synthetic route. The apparent partition coefficient of 5-ASA-Tau in 1-octanol/pH 6.8 phosphate buffer or $CHCl_3$/pH 6.8 phosphate buffer was 0.10 or 0.18, respectively, at $37^{\circ}C$. To determine the chemical and biochemical stability in the upper intestinal environment, 5-ASA-Tau was incubated in pH 1.2 and 6.8 buffer solutions, and with the homogenates of tissue and contents of stomach or small intestine of rats at $37^{\circ}C$. 5-ASA was not detected from any of the incubation medium with no change in the concentration of 5-ASA-Tau. On incubation of 5-ASA-Tau with the cecal and colonic contents of rats, the fraction of the dose released as 5-ASA was 45% and 20%, respectively, in 8 h. Considering low partition coefficient and stability in the upper intestine, 5-ASA-Tau might be nonabsorbable and stable in the upper intestine. After oral administration, it would be delivered to the colon in intact form and release 5-ASA and taurine. These results suggested 5-ASA-Tau as a promising colon-specific prodrug of 5-ASA.
Two experiments were carried out to investigate whether duckweed is useful as a dietary protein source for fine-wool Merino sheep and to evaluate its effects on wool yield and characteristics. In Experiment 1, the sheep were given one of three maintenance diets consisting of oaten chaff (520-700 g/d) supplemented with 16-32 g crude protein/d in the form of fresh (1 kg/d) or sun-dried (50-100 g/d) duckweed. Each ration was estimated to provide 5.4 MJ (1.3 Mcal)/d of metabolisable energy (ME). The sheep readily ingested the fresh or dried duckweed. None of the wool measures (yield, rate of fibre elongation, fibre diameter) differed (p>0.05) between dietary treatments. In Experiment 2, oaten-chaff-based diets (800 g/d) supplying 6.5-7.2 MJ (1.6-1.7 Mcal)/d of ME were supplemented with iso-nitrogenous amounts (4-5 g N) either of urea (8 g), cottonseed meal (60 g) or dried duckweed (100 g). In this experiment, the rate of wool fibre elongation, thought to be related to intestinal amino acid absorption, was lower (p<0.05) for sheep given the oaten chaff/urea diet than for those given either oaten chaff/cottonseed meal or oaten chaff/duckweed for which the rates did not differ (p>0.05). Fibre diameter, which ranged from 16.0-16.7 mm, did not differ (p>0.05) between diets, but tended to be lower on the oaten chaff/urea diet so that volume of wool produced was also significantly lower (p<0.05) on this diet than on the diets containing duckweed or cottonseed meal. Rumen ammonia concentrations at 4.5 and 7.5 h after feeding were higher (p<0.05) for sheep given the oaten chaff/urea diet than for those given the other two diets. A comparison of the rumen ammonia concentrations, wool growth rate and predicted flows of amino acids from the rumen of sheep supplemented with duckweed rather than cottonseed meal suggested that duckweed is a valuable source of 'escape protein' for ruminants.
Seong, Myeong A;Woo, Jong Kyu;Kang, Ju-Hee;Jang, Yeong Su;Choi, Seungho;Jang, Young Saeng;Lee, Taek Hwan;Jung, Kyung Hoon;Kang, Dong Kyu;Hurh, Byung Seok;Kim, Dae Eung;Kim, Sun Yeou;Oh, Seung Hyun
BMB Reports
/
제48권7호
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pp.419-425
/
2015
Ginseng has been widely used for therapeutic and preventive purposes for thousands of years. However, orally administered ginseng has very low bioavailability and absorption in the intestine. Therefore, fermented ginseng was developed to enhance the beneficial effects of ginseng in the intestine. In this study, we investigated the molecular mechanisms underlying the anti-inflammatory activity of fermented wild ginseng (FWG). We found that FWG significantly alleviated the severity of colitis in a dextran sodium sulfate (DSS)-induced colitis mouse model, and decreased expression level of pro-inflammatory cytokines in colonic tissue. Moreover, we observed that FWG suppressed the infiltration of macrophages in DSS-induced colitis. FWG also attenuated the transcriptional activity of nuclear factor-κB (NF-κB) by reducing the translocation of NF-κB into the nucleus. Our data indicate that FWG contains anti-inflammatory activity via NF-κB inactivation and could be useful for treating colitis. [BMB Reports 2015; 48(7): 419-425]
Purpose: This study was conducted to optimize the conditions of manufacturing the wet noodle added with Aloe powder. Methods: The I-optimal design of response surface methodology (RSM) was used for the optimization of added amount of Aloe powder, water content, and kneading time as the independent variables. The quality characteristics (color, texture, water absorption ratio, volume, turbidity) and sensory characteristics (color, flavor, taste, mouth feel, overall acceptance) were analyzed as dependent variables. Results: The lightness, redness, and yellowness of the Aloe noodle were all lowered with the addition of Aloe powder. All texture characteristics such as hardness, elasticity and chewiness of Aloe noodle showed the tendency to increase with increasing amount of Aloe added and kneading time. The water uptake and the volume expansion tended to increase with increasing aloe addition and water addition. As the kneading time increased, the dissolution of solids decreased and the turbidity tended to decrease. Aloe powder content influenced the color, flavor and taste of the noodles most among independent variables, and the mouthfeel of the noodle influenced by the water addition and the kneading time. Conclusion: Aloe noodle showed the best desirability with 3.03% of Aloe powder, 43.56% of water content and 13.06 min of kneading time by RSM analysis. Aloe noodles prepared under these optimized conditions are expected to be able to manufacture and utilize functional Aloe noodles by meeting the content of isobarbaloin, which helps the intestinal functional activity.
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