Intestinal Amino Acid Transport Research Articles

Optimal glycine allowance levels in low-protein diets and the dynamic requirement model for broilers

This study aimed to investigate the effects of different glycine levels in low-protein diets on the growth, nitrogen deposition, and expression of intestinal amino acid and glucose transporters in broilers from 29 to 42 days of age, in order to determine the optimal glycine supplementation level. A total of 240 male broilers at 29 days old were randomly assigned to 5 groups: the control group with a crude protein level of 20%, and experimental groups with low-protein diets (LP130) containing 18% crude protein, supplemented with glycine to achieve standardized ileal digestible (SID) glycine + serine to lysine ratios of 134% (LP134), 140% (LP140), and 145% (LP145). The results showed that the LP134 group had similar growth performance and slaughter performance compared to the control group (P > 0.05), whereas other low-protein diet groups had significantly lower growth performance (P < 0.05). Regression analysis determined that the optimal ratio for SID glycine + serine to lysine was 137%. A dynamic model for glycine + serine requirements was established through binary regression analysis: y = 599.051 × BW^0.75 + 8.381 × ADG (R2 = 0.998, P < 0.001). Feeding LP134, LP140, and LP145 diets significantly improved nitrogen deposition rates in broilers (P < 0.05). Low-protein diets significantly upregulated mRNA levels of b0,+AT, EAAT3, and SGLT1 genes in the duodenum (P < 0.05). In conclusion, appropriate glycine supplementation in low-protein diets can enhance growth performance, and nitrogen deposition efficiency, and regulate the expression of intestinal amino acid and glucose transporters. The optimal ratio of SID glycine + serine to lysine in low-protein diets for broilers aged 29 to 42 days is 137%.

The effect of reducing dietary energy on performance, intestinal morphology and intestinal peptide and amino acid transporters in broiler chicks

AbstractBackgroundPrevious research has investigated the impact of different factors such as gender and age of birds, protein and amino acid levels, protein origin and the physical form of the diet, on the mRNA expression of intestinal peptides and amino acid transporters. Nevertheless, there have been no attempts to examine the impact of dietary energy density on the mRNA abundance of intestinal amino acid and peptide transporters in humans or animals. This is the first reported case in both humans and animals.ObjectivesThis experiment aimed to evaluate the influence of two levels of metabolizable energy ([ME]; 2950 and 2850 kcal ME/kg diet) on performance and jejunal morphology and mRNA expression of amino acid (B0AT, b0,+AT, CAT1 and y+LAT1) and peptide (PepT1) transporters in broiler chickens during the starter period (0–10 days of age).MethodsTwo hundred and seventy day‐old male Ross 308 broiler chickens were randomly assigned to two dietary treatments with nine replicates and fifteen birds in each replicate. At 10 days of age, the outcome variables were measured, which included the assessment of the mRNA abundance of PepT1, b0,+AT, B0AT, y+LAT1 and CAT1 in the jejunal section using real‐time PCR.ResultsThe findings of this study revealed no significant impact of the dietary treatments on feed intake, body weight gain, feed conversion ratio and the jejunal mRNA abundance of PepT1, CAT1 and y+LAT1 (p &gt; 0.05). However, a reduction in dietary ME level from 2950 to 2850 kcal ME/kg diet resulted in a decrease in the mRNA level of B0AT and b0,+AT (p ≤ 0.05). The reduction in dietary energy level did not lead to any significant changes in villus width, villus height, crypt depth, villus surface area and villus height to crypt depth ratio (p &gt; 0.05).ConclusionThe results demonstrated that decreasing dietary energy density may alter the expression of amino acid transporters in the intestine while having no impact on the growth performance and morphometric structure of the intestine.

Involvement of ACE2 in the intestinal transport of amino acids: Possible health and nutritional consequences in altered expression

Background:Absorption of neutral amino acids across the intestinal cells is mediated by the broad neutral amino acid transporter B0AT1. Intestinal expression of B0AT1 depends on the coexistence with the membrane-attached angiotensin-converting enzyme 2 (ACE2) which binds to the B0AT1 or the sodium-dependent imino-acid transporter 1 (SIT1) in the intestinal epithelial cells. Objectives:The objective of this article is to provide an overview of the involvement of ACE2 in the transport of amino acids from the intestinal lumen and possible harmful effects on health and nutrition due to altered ACE2 expression. Methods: PubMed, Scopus, and Google Scholar were searched using the keywords ACE2 paired with intestinal and gut B0AT1, SIT1, amino acid transport or absorption, health, and nutrition. Expression of B0AT1 in the intestine was completely absent in ACE2 knockout mice. Lack of ACE2 and B0AT1 expression lead to low plasma and muscle tryptophan, niacin deficiency, decreased intestinal antimicrobial peptide, and increased susceptibility to inflammatory bowel disease in animal models. Results: Expression of ACE2 in the small intestine and its association with amino acid transporters play a crucial role in the absorption of amino acids, maintenance of structural and functional gut integrity and overall health and nutrition status of an individual. Bangladesh Journal of Medical Science Vol. 22 No. 04 October’23 Page : 729-733

Intestinal Amino Acid Transport and Metabolic Health.

Amino acids derived from protein digestion are important nutrients for the growth and maintenance of organisms. Approximately half of the 20 proteinogenic amino acids can be synthesized by mammalian organisms, while the other half are essential and must be acquired from the nutrition. Absorption of amino acids is mediated by a set of amino acid transporters together with transport of di- and tripeptides. They provide amino acids for systemic needs and for enterocyte metabolism. Absorption is largely complete at the end of the small intestine. The large intestine mediates the uptake of amino acids derived from bacterial metabolism and endogenous sources. Lack of amino acid transporters and peptide transporter delays the absorption of amino acids and changes sensing and usage of amino acids by the intestine. This can affect metabolic health through amino acid restriction, sensing of amino acids, and production of antimicrobial peptides.

Glucose supplementation improves intestinal amino acid transport and muscle amino acid pool in pigs during chronic cold exposure.

Mammals in northern regions chronically suffer from low temperatures during autumn-winter seasons. The aim of this study was to investigate the response of intestinal amino acid transport and the amino acid pool in muscle to chronic cold exposure via Min pig models (cold adaptation) and Yorkshire pig models (non-cold adaptation). Furthermore, this study explored the beneficial effects of glucose supplementation on small intestinal amino acid transport and amino acid pool in muscle of cold-exposed Yorkshire pigs. Min pigs (Exp. 1) and Yorkshire pigs (Exp. 2) were divided into a control group (17°C, n=6) and chronic cold exposure group (7°C, n=6), respectively. Twelve Yorkshire pigs (Exp. 3) were divided into a cold control group and cold glucose supplementation group (8°C). The results showed that chronic cold exposure inhibited peptide transporter protein 1 (PepT1) and excitatory amino acid transporter 3 (EAAT3) expression in ileal mucosa and cationic amino acid transporter-1 (CAT-1) in the jejunal mucosa of Yorkshire pigs (P<0.05). In contrast, CAT-1, PepT1 and EAAT3 expression was enhanced in the duodenal mucosa of Min pigs (P<0.05). Branched amino acids (BCAA) in the muscle of Yorkshire pigs were consumed by chronic cold exposure, accompanied by increased muscle RING-finger protein-1 (MuRF1) and muscle atrophy F-box (atrogin-1) expression (P<0.05). More importantly, reduced concentrations of dystrophin were detected in the muscle of Yorkshire pigs (P<0.05). However, glycine concentration in the muscle of Min pigs was raised (P<0.05). In the absence of interaction between chronic cold exposure and glucose supplementation, glucose supplementation improved CAT-1 expression in the jejunal mucosa and PepT1 expression in the ileal mucosa of cold-exposed Yorkshire pigs (P<0.05). It also improved BCAA and inhibited MuRF1 and atrogin-1 expression in muscle (P<0.05). Moreover, dystrophin concentration was improved by glucose supplementation (P<0.05). In summary, chronic cold exposure inhibits amino acid absorption in the small intestine, depletes BCAA and promotes protein degradation in muscle. Glucose supplementation ameliorates the negative effects of chronic cold exposure on amino acid transport and the amino acid pool in muscle.

Agouti overexpression in a transgenic model regulates integrity, permeability and electrogenic amino acid transport in zebrafish intestine

Overexpression of asip1 in transgenic zebrafish disrupts dorsoventral pigment pattern in addition to increasing food intake levels and linear growth. A higher feed intake is unnecessary in transgenic fish to enable larger and heavier growth. A plausible explanation may rely on the enhanced feeding efficiency mediated by improved nutrient absorption in transgenic animals. To test this hypothesis, wide scope transcriptomic techniques were used to elucidate the potential pathways involved in the enhanced nutrient absorption and intestinal epithelium permeability/integrity. In addition, the electrogenic capacity for amino acid transport was analysed. Transcriptomic analysis reveal that amino acid, monocarboxylates, ionic and vitamin transmembrane transporters were substantially modified. Enrichment analysis also revealed an inhibition of intestinal lipid metabolism and down-regulation of KEGG pathways related to membrane integrity suggesting augmented intestinal laxity that may enhance paracellular transport. Electrophysiological experiments carried out in Ussing chambers show that asip1 overexpression decrease membraned tissue resistance (Rt), indicating a modification of the intestinal barrier function in ASIP1 transgenic animals. Similarly, paracellular permeability was higher in transgenic zebrafish. Both the decrease in Rt and the increase in permeability point to an ASIP1-dependent decrease in the tissue barrier function. Electrogenic amino acid transport was also enhanced in transgenic animals providing strong indication that ASIP1 fish can extract more amino acids from their diet at similar feeding levels. Both transcriptomic and electrophysiological results suggest that asip1-overexpressing zebrafish display improved nutrient absorption and by extension a higher feed efficiency which explains enhanced growth in the absence of augmented food intake. The enhanced growth of ASIP1 zebrafish potentially mediated by improved nutrient uptake and feed efficiency suggests that the melanocortin system, specifically asip1 overexpression, is a potential target for the development of genetically engineered fish displaying improved performance and no differential lipid accumulation.

The beneficial effects of exogenous protease K originated from Parengyodontium album on growth performance of grass carp (Ctenopharyngodon idella) in relation to the enhanced intestinal digestion and absorption capacities

The study was primarily designed to analyze the effect of exogenous protease K on growth performance, amino acid metabolism, digestion and absorption capacities, expression of amino acid transporters related genes and proteins in intestine of sub-adult grass carp (Ctenopharyngodon idella). Six experimental diets with protease K (0, 3000, 6000, 9000, 12,000 and 15,000 U/kg) were fed to 450 grass carp (619.51 ± 0.54 g) for 60 days. The results indicated that the highest value of final body weight, percent weight gain (PWG), specific growth rate, protein efficiency ratio, activity of hepatopancreas glutamate-oxaloacetate transaminase and glutamate-pyruvate transaminase, and the lowest concentration of plasma ammonia were observed in groups with protease K of 6000 and 9000 U/kg. Meanwhile, the groups with 6000 and 9000 U/kg protease K exhibited the highest activities of digestive and brush border membrane enzymes in the three intestinal segments, as well as the highest value of intestinal somatic index and intestinal length index. The highest concentration of free amino acid in the serum and the lowest in intestine were observed in groups with supplementation of protease K 6000 and 9000 U/kg, which might be related to the up-regulated mRNA level and protein expression of intestinal amino acid transporters (SLC1A5, SLC7A6, SLC1A3 et al) in these groups. Furthermore, optimal levels of protease K up-regulated the mRNA levels of target of rapamycin (TOR), S6 kinase 1 (S6K1), and protein expression of TOR and phosphorylation TORSer2448, but reduced the mRNA level of 4E binding protein 1 (4EBP1). In summary, optimal levels of exogenous protease K was found to be conducive to facilitate the improvement of growth performance probably via enhancing digestion and absorption capacity, and utilization of amino acid in sub-adult grass carp. In addition, using quadratic regression analysis of PWG and alkaline phosphatase in the midgut, the optimum addition of protease K for the sub-adult grass carp were noted to be 7380.82 and 7406.44 U/kg, respectively.

Dietary protein hydrolysate effects on growth, digestive enzymes activity, and expression of genes related to amino acid transport and metabolism of larval snakehead (Channa argus)

A feeding trial was conducted to investigate the effects of dietary protein hydrolysate on the growth, digestive enzyme activity, and expression of genes related to amino acid transport and metabolism in larval snakehead. Five isonitrogenous (crude protein, 59%) and isolipidic (crude lipid, 13%) diets were formulated using pre-mixed protein hydrolysates to replace approximately 0% (control, PH0), 25% (PH25), 50% (PH50), 75% (PH75) and 100% (PH100) un-hydrolyzed pre-mixed protein. Triplicate groups of snakehead larvae (17 days post-hatching, initial body weight 122 ± 0.10 mg) were randomly assigned to 15 fiberglass fish tanks with a density of 750 larvae per tank, and fed four times daily for 20 days. At the end of the feeding trial, the growth metric was monitored, and tissues, including intestine and muscle, were separated for physiology and molecular biology analysis. The results showed that dietary protein hydrolysate quadratically increased final body weight (FBW, 2.57–3.73 g) and survival rate (SR, 53.33–60.14%), with the maximum value being observed in the PH75 group (FBW, 3.73 g; SR, 60.14%). Meanwhile, the optimal level of dietary protein hydrolysate elevated the activity of digestive enzymes including trypsin (1558.24–2520.37 U/gpro), lipase (22.37–36.26 U/gpro) and alkaline phosphatase (3.05–3.98 U/mgpro). Gene expression results revealed that protein hydrolysate inclusion quadratically elevated the expression of intestinal amino acid transporter y+LAT2 (0.96–1.51) and peptide transporter PepT1 (0.68–1.22), with the maximum value observed in the PH75 (1.51) and PH50 (1.22) groups, respectively. Meanwhile, the expression of muscle PepT1 (0.97–1.73) was up-regulated linearly, and y+LAT2 expression (0.54–1.01) quadratically decreased with the inclusion of protein hydrolysate. Dietary protein hydrolysate appeared activate the target of rapamycin (TOR) pathway in the muscle through the up-regulation of TOR expression (1.00–1.20) and down-regulation of 4EBP1 expression (0.56–1.01). Additionally, the expression of genes involved in the amino acids response (AAR) pathway in muscle, eIF2α (0.50–1.03), CHOP (0.36–1.09) and REDD1 (0.27–0.98), were linearly inhibited with the inclusion of protein hydrolysate. In above, the optimal level of dietary protein hydrolysate (75%) elevated the growth performance and improved the digestive tract of snakehead larvae. The beneficial role of protein hydrolysates in growth and development is partly attributed to the regulation of genes related to amino acid transport and metabolism.

Functional Properties of Protein Hydrolysates on Growth, Digestive Enzyme Activities, Protein Metabolism, and Intestinal Health of Larval Largemouth Bass (Micropterus salmoides).

The present study was conducted to investigate the effects of dietary inclusion of protein hydrolysates on growth performance, digestive enzyme activities, protein metabolism, and intestinal health in larval largemouth bass (Micropterus salmoides). The experimental feeding trial presented in this study was based on five isonitrogenous and isolipidic diets formulated with graded inclusion levels of protein hydrolysates, and it showed that protein hydrolysates improved growth performance, reduced larval deformity rate, and increased the activity of digestive enzymes, including pepsin and trypsin. Gene expression results revealed that the supplementation of protein hydrolysates upregulated the expression of intestinal amino acid transporters LAT2 and peptide transporter 2 (PepT2), as well as the amino acid transporters LAT1 in muscle. Dietary provision of protein hydrolysates activated the target of rapamycin (TOR) pathway including the up-regulation of TOR and AKT1, and down-regulation of 4EBP1. Additionally, the expression of genes involved in the amino acids response (AAR) pathway, ATF4 and REDD1, were inhibited. Protein hydrolysates inhibited the transcription of some pro-inflammatory cytokines, including IL-8 and 5-LOX, but promoted the expression of anti-inflammatory cytokines TGF-β and IL-10. The 16S rRNA analysis, using V3-V4 region, indicated that dietary protein hydrolysates supplementation reduced the diversity of the intestine microbial community, increased the enrichment of Plesiomonas and reduced the enrichment of Staphylococcus at the genus level. In summary, protein hydrolysates have been shown to be an active and useful supplement to positively complement other protein sources in the diets for largemouth bass larvae, and this study provided novel insights on the beneficial roles and possible mechanisms of action of dietary protein hydrolysates in improving the overall performance of fish larvae.

Dietary tea tree (Melaleuca alternifolia) oil supplementation enhances the expressions of amino acid transporters in goat ileal mucosa and improves intestinal immunity.

Tea tree oil (TTO) is a plant-derived additive with anti-inflammatory, bactericidal, and growth-promoting properties. However, little is known about the effects of TTO on intestinal amino acid transport and immune function in goats. Twenty-four Ganxi goats (initial body weight of 13.5 ± 0.70 kg) were randomly allotted two treatments and fed either control (CON) or CON+TTO (0.2ml/kg) diet. The addition of TTO to the diet significantly decreased (p < .05) tumor necrosis factor-α content and increased (p < .05) interleukin-2 (IL-2) content in goat serum; significantly decreased (p < .05) IL-12, and increased (p < .05) IL-2 content in goat ileal mucosa; significantly increased (p < .05) secreted IgA content in the jejunal and ileal mucosa; significantly upregulated (p < .05) IL-2 and downregulated (p < .05) IL-12 at the mRNA level in the ileal mucosa; significantly elevated the levels of serine, arginine, and total amino acids in the ileal mucosa (p < .05); significantly upregulated (p < .05) SLC1A1 and SLC7A1 in the ileum; and significantly enhanced (p < .05) the protein expression of Claudin-1 in the ileal mucosa. In summary, adding 0.2 ml/kg of TTO to the diet enhanced SLC1A1 and SLC7A1 mRNA expression in the ileal mucosa, and SLC1A1 and SLC7A1 could transport serine and arginine from the chyme to the ileal mucosa. Thus, increased serine and arginine content in the mucosa could improve intestinal immunity. TTO supplementation upregulated the expression of IL-2 and Claudin-1 in goat ileal mucosa, and enhanced immune function in the intestine.

P73. Expression of intestinal amino acid transporters, gut morphology, and amino acid digestibility in pigs fed proteins or free amino acids

P73. Expression of intestinal amino acid transporters, gut morphology, and amino acid digestibility in pigs fed proteins or free amino acids

Changes to gut amino acid transporters and microbiome associated with increased E/I ratio in Chd8+/\u2212 mouse model of ASD-like behavior

Autism spectrum disorder (ASD), a group of neurodevelopmental disorders characterized by social communication deficits and stereotyped behaviors, may be associated with changes to the gut microbiota. However, how gut commensal bacteria modulate brain function in ASD remains unclear. Here, we used chromodomain helicase DNA-binding protein 8 (CHD8) haploinsufficient mice as a model of ASD to elucidate the pathways through which the host and gut microbiota interact with each other. We found that increased levels of amino acid transporters in the intestines of the mouse model of ASD contribute to the high level of serum glutamine and the increased excitation/inhibition (E/I) ratio in the brain. In addition, elevated α-defensin levels in the haploinsufficient mice resulted in dysregulation of the gut microbiota characterized by a reduced abundance of Bacteroides. Furthermore, supplementation with Bacteroides uniformis improved the ASD-like behaviors and restored the E/I ratio in the brain by decreasing intestinal amino acid transport and the serum glutamine levels. Our study demonstrates associations between changes in the gut microbiota and amino acid transporters, and ASD-like behavioral and electrophysiology phenotypes, in a mouse model.

Dietary lysine supplementation improves growth performance and skeletal muscle development in rabbits fed a low protein diet

The purpose of this study was to investigate the effects on growth of Lysine (Lys) supplementation in a low protein diet. We also investigated the gene or protein expression related to skeletal muscle development and intestinal amino acid transporters, and determined the major signalling associated with Lys-regulating skeletal muscle development. 1000healthy, weights averaging 938.6±6.54g weaned rabbits were randomly divided into five groups (five replicates in each group and 40 rabbits in each replicate). These groups consisted of the normal protein group (NP group, consuming a diet containing 16.27% protein), the low protein group (LP group, 14.15%-14.19% protein) and the LP group with an addition of 0.15%, 0.3% or 0.45% Lys. The trial included 7 d of pre-feeding and 28 d of exposure to the treatment. Compared with NP diet and LP diet, LP+0.3% Lys group improved growth performance (p<0.05), full-bore weight and half-bore weight of rabbits (p<0.05). The LP+0.3% Lys group also resulted in a decrease in the excretion of faecal nitrogen and urinary nitrogen (FN; UN; p<0.05), and an increase in nitrogen utilisation rate (NUR; p<0.05). LP diet increased the mRNA expression of MSTN and WWP1, and decreased the mRNA expression of IGF1 (p<0.05). LP diet decreased the protein expression of P-P70S6K1, P-4EBP1 and P-S6 (p<0.05). LP+0.3% Lys group attenuated the effects of LP diet on the expression of MSTN, WWP1, IGF1, P-P70S6K1, P-4EBP1 and P-S6 (p<0.05). LP+0.3% Lys group resulted in an increase in mRNA expression of MyoD and protein expression of P-mTOR relative to the NP and LP groups (p<0.05). In summary, the addition of Lys to a LP diet provides a theoretical basis for the popularisation and application of Lys in rabbit production.

Dietary amino acid supplementation affects temporal expression of amino acid transporters and metabolic genes in selected and commercial strains of rainbow trout (Oncorhynchus mykiss).

Replacement of fishmeal as the major protein source in feeds is critical for continued growth and sustainability of the aquaculture industry. However, numerous studies have shown suboptimal fish growth performance and reduced protein retention efficiency when carnivorous fish species are fed low fishmeal-high plant protein feeds. A study was conducted using a commercial strain and a genetically improved strain of rainbow trout selected for improved performance when fed an all plant protein diet to identify physiological differences associated with growth performance in the selected trout strain. Fifty individuals per strain (average weight~580g) were force-fed a plant-protein blend with and without amino acid supplementation (lysine, methionine and threonine) at 0.5% body weight and sampled at intervals over 24h. Samples from intestine and liver were analyzed for specific gene expression analysis related to amino acid transporters, digestive process control, protein degradation and amino acid metabolism. The results showed that expression levels of various intestinal amino acid transporters (SLC1A1, SLC7A9, SLC15A, SLC1A5 SLC6A19 and SLC36A1) were affected by strain, diet and time. Moreover, significant interactions were found regarding the temporal expression levels of cholecystokinin (CCK-L), Krüppel-like factor 15 (KLF15) and aspartate aminotransferase (GOT) transcripts in the examined tissues. The results provide evidence that improved growth and protein retention of the selected strain fed an all-plant protein diet is a result of nutritional adaptation and an overall change in physiological homeostatic control.

Addition of hydrolysed porcine mucosa to low-fishmeal feed improves intestinal morphology and the expressions of intestinal amino acids and small peptide transporters in hybrid groupers (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂)

Aquaculture routinely uses fishmeal (FM) as feed, but its increasing cost has prompted the search for alternative protein sources. This study conducted an 8-week feeding trial to evaluate the effects of adding hydrolysed porcine mucosa (HPM) to low-FM feed on the growth performance, intestinal morphology, and expressions of intestinal amino acids and small peptide transporters in hybrid groupers (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂, initial weight 7.51 ± 0.03 g). Five iso-nitrogenous and isolipidic experimental diets were formulated containing HPM in proportions of 0% (Group HPM0), 3% (HPM3), 6% (HPM6), 9% (HPM9) and 12% (HPM12). Specific growth and weight gain rates were significantly lower in fish fed >6% HPM (P < 0.05). Catalase and glutamic oxaloacetic transaminase activities were significantly higher in Group HPM12 than in Group HPM0 (P < 0.05). In the distal intestine (DI), Group HPM6 had significantly higher villus length than Group HPM3, and significantly higher muscle thickness than Groups HPM0, HPM3 and HPM12 (P < 0.05). The alpha diversity index of intestinal microbiota was not significantly affected by HPM content (P > 0.05). Expressions of LAAT1 in the proximal intestine (PI), CAT2, SNAT3 and EAAT1 in the mid intestine (MI) and CAT2, B0AT3 and EAAT1 in the DI were highest in Group HPM3 (P < 0.05). Expressions of CAT1, CAT2, SNAT3 and EAAT1 in the PI, CAT1, PAT1, B0AT2 and B0AT3 in the MI, and PAT1 and SNAT3 in the DI were highest in Group HPM6 (P < 0.05). The expressions of PepT1 of the MI and DI were significantly higher in Groups HPM9 and HPM12 than in others (P < 0.05). The results show that adding 3% or 6% HPM to feed can improve DI morphology and increase intestinal amino acid transporter expression without reducing growth performance. Dietary supplementation with 12% HPM may improve antioxidant capacity and expression of small peptide transporters but may also cause liver damage and reduce growth performance. Therefore, HPM is a promising alternative to FM for use in aquaculture feeds.

Impact of soy lecithin, zinc oxide, and methylsulfonylmethane, as excipient ingredients, on the bioaccessibility and intestinal transport of branched-chain amino acids from animal and plant protein mixtures.

To maximize the biological activity of branched-chain amino acids (BCAAs), it is necessary to find a new excipient agent to increase the bioavailability of BCAAs in protein mixtures. The aim of the current study was to investigate the effects of soy lecithin (SLC), zinc oxide (ZnO), and methylsulfonylmethane (MSM) on the bioaccessibility and intestinal transport of BCAAs from animal and plant protein mixtures (PMs) via an in vitro digestion model with human intestinal epithelial (Caco-2) cells. The bioaccessibility of total BCAAs in PMs considerably increased by 107.51 ± 1.50% with the addition of SLC, and the combined effects of SLC, ZnO, and MSM on enhancing the bioaccessibility of total BCAAs was observed (107.14 ± 0.18%). Interestingly, SLC showed a major role in binding bile acid, showing 65.78 ± 1.66% of binding capacity. Intestinal transport of BCAAs was measured to be at 100.48, 110.86, and 130.29 μg mL-1 for leucine, isoleucine, and valine, respectively, in PMs with SLC + ZnO + MSM, and it eventually amplified the amount of the total transported BCAAs (341.63 ± 6.34 μg mL-1), which was about 8.72 times higher than that of PM only. The cellular integrity of digesta-treated Caco-2 cells tended to decrease according to the incubation time, but it was recovered in the treatment of PM + SLC + ZnO + MSM, and nearly reached the control levels with 92.82 ± 0.53%. Results from the current study suggest that the co-consumption of proteins equally consisting of plant and animal sources with SLC, ZnO, and MSM could improve the bioavailability of total BCAAs, resulting in the improvement of health benefits.

Plasma citrulline correlates with basolateral amino acid transporter LAT4 expression in human small intestine

Background & aimsPlasma citrulline, a non-protein amino acid, is a biochemical marker of small intestine enterocyte mass in humans. Indeed, citrulline is highly correlated with residual bowel length in patients with short bowel syndrome. It is known to be synthesised in epithelial cells of the small intestine from other amino acids (precursors). Citrulline is then released into systemic circulation and interconverted into arginine in kidneys. If plasma citrulline concentration depends on abundance of intestinal amino acid transporters is not known. The aim of the present study was to explore whether plasma citrulline concentration correlates with the expression of intestinal amino acid transporters. Furthermore, we assessed if arginine in urine correlates with plasma citrulline.MethodsDuodenal samples, blood plasma and urine were collected from 43 subjects undergoing routine gastroduodenoscopy. mRNA expression of seven basolateral membrane amino acid transporters/transporter subunits were assessed by real-time PCR. Plasma and urine amino acid concentrations of citrulline, its precursors and other amino acids were analysed using High Performance Liquid Chromatography measurements. Amino acid transporter mRNA expression was correlated with blood plasma and urine levels of citrulline and its precursors using Spearman's rank correlation. Likewise, urine arginine was correlated with plasma citrulline.ResultsPlasma citrulline correlated with the mRNA expression of basolateral amino acid transporter LAT4 (Spearman's r = 0.467, p = 0.028) in small intestine. None of the other basolateral membrane transporters/transporter subunits assessed correlated with plasma citrulline. Plasma citrulline correlated with urinary arginine, (Spearman's r = 0.419, p = 0.017), but not with urinary citrulline or other proteinogenic amino acids in the urine.ConclusionsIn this study, we showed for the first time that small intestinal basolateral LAT4 expression correlates with plasma citrulline concentration. This finding indicates that LAT4 has an important function in mediating citrulline efflux from enterocytes. Furthermore, urine arginine correlated with plasma citrulline, indicating arginine in the urine as possible additional marker for small intestine enterocyte mass. Finally, basolateral LAT4 expression along the human small intestine was shown for the first time.

Dietary Insect Powder Protein Sources Improve Protein Utilization by Regulation on Intestinal Amino Acid-Chemosensing System.

Simple SummaryInsect powders, including Tenebrio molitor (TM), Musca domestica larvae (MDL) and Zophobas morio (ZM), as high-quality and renewable protein sources are commonly applied in livestock and poultry feed production. The molecular effect of insect protein on amino acid metabolism in pigs needs to be explored. We found that insect powder as a protein source in feed regulated the mTOR signal pathway and improved amino acid transportation in the intestine for growth promotion. Insect powder may be a potentially promising protein source for pig production.This study was conducted to evaluate the effects of dietary insect powder supplementation as a protein source on plasma amino acid profiles, intestinal amino acid transport and sensing in a piglet model. A total of 144 weanling piglets were randomly assigned to four experimental diets for two phases (Days 1–28 and Days 29–56), to assess the effects on amino acid profiles and transportation in the segments of the intestine. The groups were basal diet (control), control diet plus Tenebrio molitor (TM), control diet plus Musca domestica larvae (MDL) and control diet plus Zophobas morio (ZM). The plasma free amino acid levels were stable comparable among treatments, except that the lysine level was significantly reduced by dietary MDL and ZM supplementation in the first phase (p < 0.05). In the 1st phase, the sensitivity of intestinal segments to the regulation of the amino acid level by insect powder supplementation follows sequence: colon > ileum > jejunum, while the order switched to jejunum > colon > ileum in the 2nd phase. The relative RNA expressions of mitogen-activated protein 4 kinase 3 (MAP4K3), sodium dependent neutral amino acid transporter2 (SNAT2), the transient receptor potential cation channel subfamily V member 1 (TRPV1) and taste 1 receptor member 1/3 (T1R3) in the segments of the intestine were affected by different dietary insect powder supplementation. G protein-coupled receptor family C group 6 member A (GPRC6A) level in the jejunal and colonic mucosa was upregulated by MDL supplementation (p < 0.05). These results indicated that dietary insects improved the metabolism of the amino acid in the prophase (the 1st phase) through regulating the sensing gene and mTOR signal pathway in intestinal mucosa by targeting different receptors. The finding demonstrates that the insect powder is a potentially promising source for protein deposition.

Branched-chain amino acids, especially of leucine and valine, mediate the protein restricted response in a piglet model.

Branched-chain amino acids (BCAAs) are reduced in various protein restricted models, while the detailed role of BCAAs in protein restricted response is still obscure. Thus, the current study mainly investigated the amino acid metabolism in protein restricted piglets and the effects of BCAA balance in a low-protein diet on growth performance, amino acid metabolism, intestinal structure, and gut microbiota with focus on which BCAAs contributed to the protein restricted response. The results showed that protein restriction increased serum Ser, Thr, Ala, Lys, and Trp but reduced His, Cys, Val, and Ile levels. Intestinal amino acid transporters mainly mediated the mechanism of amino acid uptake. The BCAA balance refreshed the serum BCAA pool, which further improved growth performance in protein restricted piglets. Leu, Val, and Ile balances increased serum BCAA concentrations, respectively, and Leu and Val but not Ile enhanced the feed intake and weight gain in protein restricted piglets. In addition, protein restriction impaired the villus structure and increased the number of goblet cells in the ileum. Also, gut microbiota (Spirochaetales, Gammaproteobacteria, Lactobacillales at the order level) were altered in protein restricted pigs, while the BCAA balance markedly improved Gammaproteobacteria, Lactobacillales, and Aeromonadales proliferation, which might mediate growth promotion and amino acid metabolism. In conclusion, protein restriction markedly affected the host amino acid metabolism (i.e., Ser, Thr, Lys, His, BCAAs). The BCAA balance (especially for supplementation with Leu and Val) improved the amino acid metabolism, growth performance, and gut microbiota communities.

Long-term effect of early antibiotic exposure on amino acid profiles and gene expression of transporters and receptors in the small intestinal mucosa of growing pigs with different dietary protein levels.

This study evaluated the effects of early antibiotic exposure (EAE) on subsequent amino acid (AA) profiles and small intestinal AA transporter and receptor expression level in pigs with different dietary crude protein (CP) levels. Eighteen litters of piglets were fed creep feed diets, either with or without antibiotics while with sow on day 7. The pigs were weaned at day 23 and fed the same diets until day 42, when random pigs within each group were offered a normal- or low-CP diet, thereby creating four groups. On day 120, the pigs were euthanized, and jejunal and ileal mucosa and digesta were collected for gene-expression and AA-concentration analysis. With the normal-CP diet, EAE increased (P < 0.05) the concentrations of six essential amino acids (EAA) and three non-essential amino acids (NEAA) in serum, four EAAs and four NEAAs in jejunal mucosa, one EAA and two NEAAs in ileal mucosa, five EAAs and three NEAAs in jejunal digesta, and three EAAs and two NEAAs in ileal digesta. Early antibiotic exposure upregulated (P < 0.05) CAT1, ASCT2, ATB0,+ , CaSR, T1R1, and T1R3 expression in the jejunum, downregulated PepT1 expression with a normal-CP diet. It upregulated (P < 0.05) the expressions of CAT1, ATB0,+ , ATP1A1, and T1R3 in the ileum with a normal-CP diet. These results suggest that EAE has long-term effects on AA profiles, mainly in the jejunum and serum, by increasing AA transporter expression in the intestine, and that these effects may be influenced by dietary CP levels. © 2019 Society of Chemical Industry.