L-canavanine inhibits L-arginine uptake by broiler chicken intestinal brush border membrane vesicles

1. Intestinal brush border membrane vesicles (BBMV) were prepared from 3-week-old broiler chickens. 2. Electron microscopy of the BBMV fraction showed single membrane vesicles of different sizes with no electron dense material inside. No other organelles were observed. The sucrase and maltase activities were enriched by factors of 16 and 18, respectively, in the BBMV fraction in comparison with the homogenate. On the other hand, the Na+/K+-ATPase sensitivity to ouabain was increased by a factor of 0.8. 3. The BBMV showed a maximum L-[14C]-arginine uptake (944.9 +/- 22.9 pmoles/mg protein) at 45 s and thereafter it declined slowly. In the presence of 0.5 mM L-canavanine, the L-[14C]-arginine uptake by BBMV was reduced by 43.6% at 45 s. 4. It is concluded that L-canavanine inhibits L-arginine Na+-dependent transport across the enterocyte apical membrane in a highly purified intestinal BBMV from broiler chickens.     

Influence of deoxynivalenol on the D-glucose transport across the isolated epithelium of different intestinal segments of laying hens

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 sodium D-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 of D-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 microg 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 mmol D-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 of D-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.     

High intestinal transport activity for nucleosides in cattle: a synopsis

In ruminants large amounts of nucleic acids associated with the microbial cell mass leaving the fore-stomach system via the abomasum enter the small intestine. In dairy cows this amounts to 100-200 g microbial nucleic acids per day. These nucleic acids are very efficiently digested in the small intestine whereby nucleosides were found to be the main degradation products. Therefore, this review centers mainly on the mechanisms of intestinal absorption of nucleosides and their role in nucleic acid digestion. Furthermore, metabolism of nucleosides in intestinal epithelial cells and its bearing for nucleoside absorption and salvage of nucleosides for nucleic acid synthesis in various tissues is considered. According to own studies using isolated intestinal brush-border membrane (BBM) vesicles (BBMV) from dairy cows purine and pyimidine nucleosides are transported actively by two separate Na+ co-transport systems (N1 and N2) across the bovine BBM, whereby transport activity in the small intestine decreases from proximal to distal. Guanosine and inosine appeared to be transported exclusively by N1 while thymidine and cytidine appeared to be transported exclusively by N2. Uridine and adenosine had an affinity to both transporters. In comparison to findings in man and rabbit, transport capacity (Vmax) of N1 and N2 in the BBM of cows was more than 10-fold higher. Similar findings were obtained in BBMV isolated from the small intestine of veal calves with rudimentary forestomach development in regard to nucleoside transport. Therefore, the high intestinal transport activity for nucleosides seems to be a genetically fixed property in the bovine, which is not related to a functional fore-stomach system.     

钠离子对仔猪小肠刷状缘膜囊二肽跨膜转运的影响

为研究Na 在仔猪小肠Gly-Pro跨膜转运中的作用,采用同位素示踪及体外孵育技术,观察在不同Na 浓度梯度条件下Gly-Pro在仔猪小肠刷状缘膜囊(brush border membrane vesicles,BBMV)的跨膜转运量。结果表明:(1)用K 替代Na ,并没有降低Gly-Pro在BBMV中的转运量,在孵育前20min时间内,Gly-Pro的转运量稍稍提高;(2)Na 可使BBMV对L-丙氨酸的转运量提高到4.6倍,但不影响Gly-Pro的转运;二氢骆驼蓬碱显著地降低L-丙氨酸的转运,但对Gly-Pro的转运无影响。研究提示,Gly-Pro在仔猪小肠BBMV体系中的转运与Na 不存在依赖性的关系。     

Digestibility of pentose sugars and uronic acids and their effect on chick weight gain and caecal size

1. In the first experiment D-xylose, L-arabinose, D-galacturonic and D-glucuronic acids were fed ad libitum to young chicks for 2 weeks at 200 g/kg of diet and weight gains and food consumption were recorded. 2. L-arabinose and D-xylose did not depress food consumption in the first week but prolonged feeding caused food consumption to decrease and weight gain to be adversely affected. 3. D-galacturonic acid and D-glucuronic acid caused severe growth retardation as early as the first week of feeding, primarily because of voluntary starvation. 4. Apparent metabolisable energy values for the diets were obtained when chicks were 19 to 21 d of age and were 14.04 +/- 0.52, 12.03 +/- 0.61, 11.77 +/- 1.21, 11.68 +/- 0.34 and 11.66 +/- 0.45 KJ/g for the basal diet with glucose, xylose, arabinose, galacturonic and glucuronic acids respectively. 5. True metabolisable energy values for the diets were obtained from adult cockerels and were 15.07 +/- 0.16, 13.45 +/- 0.16, 13.12 +/- 0.37, 12.29 +/- 0.26 and 12.69 +/- 0.23 KJ/g for basal diet with glucose, xylose, arabinose, galacturonic and glucuronic acids respectively. 6. In the second experiment D-galactose, D-xylose, L-arabinose, D-galacturonic and D-glucuronic acid were fed ad libitum to young chicks for 3 weeks at 50 g/kg of diet and weight gains and food consumption were recorded. 7. Chicks grew and ate well on all diets. 8. The digestibilities of sugars and uronic acids were obtained by measurement of these constituents in diets and digesta using titanium dioxide as a marker. The digestibilities were 1.000 +/- 0.0, 0.997 +/- 0.002, 0.936 +/- 0.041, 0.628 +/- 0.103, 0.588 +/- 0.059, and 0.645 +/- 0.089 for D-glucose, D-galactose, D-xylose, L-arabinose, D-galacturonic and D-glucuronic acids respectively. 9. Both at 200 and 50 g/kg dietary inclusion there was noticeable caecal fermentation from L-arabinose, D-galacturonic and D-glucuronic acid. Only at 200 g/kg dietary inclusion did D-xylose produce significant evidence of caecal fermentation.     

Intestinal glucose absorption in calves as affected by different carbohydrate sources

From numerous recent studies, it has been demonstrated that the development of the forestomach system in ruminants and thus microbial carbohydrate fermentation do not exclude the potential of the small intestines for enzymatic carbohydrate digestion and subsequent monosaccharide absorption. However, the role of regulatory nutritional factors is still under discussion. Therefore, we investigated the kinetic parameters of intestinal Na⁺‐dependent glucose absorption and SGLT1 expression using isolated brush border membrane vesicles (BBMV) from the jejunum of 10‐week‐old calves kept on either hay, concentrate or corn silage‐based diets in addition to milk replacer. While the maximal transport capacity was significantly higher for concentrate and corn silage‐fed animals, SGLT1 protein expression was highest in BBMV isolated from hay‐fed animals. This observation differs from the prevalent conception that induction of Na⁺‐dependent glucose uptake via SGLT1 is based on an increased number of transporters at the brush border membrane.     

Effect of erythromycin on L-threonine transport in rabbit jejunum in vitro

Several antibiotics characterized by different molecular structures are known to affect some intestinal activities. Some of them have been described as inhibitors of the intestinal sugar and amino acid transport with different mechanisms. Erythromycin (EM) is a macrolide antibiotic acting as a motilin agonist and thus stimulating the gastrointestinal motor activity. Since several substances which increase the motor activity of the gastrointestinal tract may produce effects on the intestinal absorption of nutrients, the present study has been carried out to determine whether erythromycin affects the L-threonine intestinal absorption. The results obtained indicate that erythromycin diminishes the L-threonine intestinal transport, probably at the mucosal border level. Two groups of experiments carried out, with Na(+)-deprived medium and ouabain-enriched medium, might indicate that erythromycin action could be due to either a direct or an indirect action on the Na(+)-dependent L-threonine transport located in the brush border.     

Influence of abomasal carbohydrates on small intestinal sodium-dependent glucose cotransporter activity and abundance in steers

Most animals adapt readily to increased supplies of carbohydrate in the intestinal lumen by increasing enzymes for degradation and increasing glucose transporter activity. However, the extent of upregulation of Na+-dependent glucose cotransporter 1 (SGLT1) activity and content in response to increased delivery of carbohydrate to the small intestinal lumen of ruminants is unclear. Therefore, an experiment was conducted to determine the effect of glucose and starch hydrolysate on the activity and abundance of SGLT1 in the small intestine of steers. In a randomized complete block design, 40 crossbred beef steers (243+/-2 kg BW) were fed 0.163 Mcal of ME/(kg BW0.75(d; W), 0.215 Mcal of ME/(kg BW0.75 x d; 2M), or 0.163 Mcal ME/(kg BW0.75 x d) and infused for 35 d into the rumen (R) or abomasum (A) with 12.6 g/(kg BW0.75 x d) of starch hydrolysate (S) or into the abomasum with 14.4 g/(kg BW0.75 x d) of glucose (G). Steers were slaughtered, and brush-border membrane vesicles were prepared from the small intestinal samples obtained from five equidistant sites along the intestine. Maltase activity in vesicles and homogenates differed with intestinal sampling site (quadratic, P < 0.001). Steers on the AG treatment yielded a greater intestinal maltase activity (38 nmol glucose x mg protein(-1) x min(-1)) compared with the AS, RS, W, or 2M treatments (34, 26, 23, and 23 nmol glucose x mg protein(-1) x min(-1) respectively [SEM = 3; P = 0.02]). Sodium-dependent glucose uptake averaged 18.4+/-3.94 pmol glucose/(mg protein x s) and was not affected by treatment, but uptake decreased distally along the intestine (P < 0.001). There was no effect of treatment on SGLT1 protein abundance, but SGLT1 protein abundance increased linearly from the duodenum to the ileum (P = 0.05). The inverse relationship between glucose uptake and SGLT1 abundance suggests that the regulation of brush border Na+-dependent glucose transport capacity is complex, involving factors other than the presence of luminal carbohydrate.     

膜电位对仔猪小肠刷状缘膜囊中二肽跨膜转运的影响

为研究膜电位与仔猪小肠刷状缘膜囊(brush border membrane vesicles,BBMV)中二肽跨膜转运之间的关系,采用膜电位诱导、体外孵育及同位素示踪技术,分别观察由缬氨霉素和羰基氰化物三氟甲氧基苯腙(carbonylcyanide p-(Tri-fluoromethoxy)phenylhydrazone,FCCP)诱导仔猪小肠BBMV产生的K 扩散电位(膜囊内负)和H 扩散电位(膜囊内正)对Gly-Pro二肽跨膜转运的影响。结果表明:在有Na 和无Na 的状况下,K 扩散电位均促进Gly-Pro二肽由膜囊外进入膜囊内部;H 扩散电位抑制Gly-Pro二肽由膜囊外进入膜囊内部。结果提示:在仔猪小肠BBMV转运体系中,Gly-Pro二肽的跨膜转运具有与非Na 的阳离子协同转运的特点。     

Effects of B-trichothecenes on luminal glucose transport across the isolated jejunal epithelium of broiler chickens

Trichothecenes are closely-related sesquiterpenoids (ring structure) with a 12, 13 epoxy ring and a variable number of hydroxyl, acetyl or other substituents. In chickens, D-glucose and amino acid absorption occurs via carrier-mediated transport. Recently, it has been observed that deoxynivalenol (DON) alters the gut function and impairs glucose and amino acid transport in chickens. The purpose of this work was to determine the effects of different B-trichothecenes [DON, Nivalenol (NIV), 15-Ac-DON and Fusarenon X (FUS X)] on intestinal carrier-mediated sodium co-transport of D-glucose in the small intestine of broiler chickens. Intestinal transport was determined by changes in the short-circuit current (Isc), proportional to ion transmembrane flux, in the middle segment of the jejunum of broilers with the Ussing chamber technique. D-glucose produced an increase of the Isc, and this effect was reverted by different B-trichothecene mycotoxins, indicating that the glucose induced Isc was altered by B-trichothecenes. The addition of glucose after pre-incubation of the tissues with B-trichothecenes had no effect (p > 0.05) on the Isc, suggesting that B-trichothecenes afflicted the Na(+)-D-glucose co-transport. However, FUX had no obvious effect on the measured parameters. It could be concluded from the present study that the glucose co-transporter activity appears to be more sensitive to DON, NIV and 15-Ac-DON suppression than by FUS X in the jejunum of broilers.     

Abundance of intestinal Na+/glucose cotransporter (SGLT1) in roe deer (Capreolus capreolus)

Introduction Based on morphological and physiological differences in their digestive systems H ofmann and S tewart (1972) classified ruminants into three feeding types: concentrate selectors (CS), grass and roughage eaters (GR) and intermediate, opportunistic, mixed feeders (IM). In evolutionary terms CS most closely resemble the original ruminants (H ofmann 1989). They have a relatively small forestomach with large openings between the different parts of the stomach. The retention time of ingesta is short and therefore less suited to optimizing cellulose digestion. They have relatively large saliva glands, producing high volumes of saliva (F ickel et al. 1998), which may facilitate the passage of soluble nutrients down their highly developed ventricular groove. The CS ruminants choose easily digestible forage, especially dicotyledons, which are rich in soluble plant cell contents (PCC, T ixier et al. 1997; D uncan et al. 1998). With evolution, the forestomach of the ruminants became larger relative to body size, and also more subdivided. These evolutionary adaptations optimized the selective retention of feed particles and increased retention time, leading to improvement of cellulose digestion. Large GR are well adapted to digest plant cell walls (PCW), i.e. structural carbohydrates (cellulose); they consequently feed mainly on monocotyledons. Roe deer (Capreolus capreolus) are very small ruminants which select concentrated feed. As energy requirements increase proportional to metabolic body weight (K leiber 1961) roe deer require relatively more energy per unit body mass for maintenance compared to that required by large ruminants (V an S oest 1994; D uncan et al. 1998). Easily digestible forage is not sufficient to satisfy the energy requirements of roe deer. It is proposed therefore that these animals may have other methods of avoiding energy losses due to microbial fermentation, either bypassing nutrients down the ventricular groove (rumen bypass) or by ruminal escape of unfermented and/or partially fermented nutrients (H ofmann 1989). The relatively high content of polyunsaturated fatty acids present in the depot fat of roe deer due to only partial rumen biohydrogenation (M eyer et al. 1998) supports the alternative hypothesis of rumen bypass or ruminal escape. Once reaching the small intestine, partially fermented or bypassed soluble carbohydrates can be further hydrolysed by maltase and sucrase to monosaccharides. These monosaccharides are then available for absorption. The intestinal sodium-dependent glucose cotransporter (SGLT1) in the brush border membrane (BBM) of the enterocytes is a stable indicator for the presence of sugars in the small intestine. The expression and activity of SGLT1 protein are regulated by the level of sugars reaching the small intestine (F erraris and D iamond 1989). Infusion of d -glucose into the intestinal lumen of ruminant sheep for 2 h stimulates the expression of SGLT1 which can be detected 4 days later in the newly formed mature enterocytes (S hirazi -B eechey et al. 1995). When dietary carbohydrate is removed, it takes 3 days before the brush border glucose uptake capacity decreases significantly from control values (F erraris 1994). This reflects the time of physiological cell renewal. In GR, such as sheep, the capacity of the intestine to absorb sugars decreases up to 500-fold during development (S hirazi -B eechey et al. 1995). This decrease in the intestinal absorptive capacity is a reflection of the decline in SGLT1 transport activity and protein abundance. It is not due to modifications in the structure of the absorptive surface and cannot be explained in terms of an age-related process (S hirazi -B eechey et al. 1991). It has been shown that there is a direct correlation between the level of monosaccharides reaching the lumen of the small intestine and the expression of functional SGLT1 protein (D yer et al. 1997). Therefore in this study we aimed to investigate the presence of functional SGLT1 in the small intestine of roe deer as a stable indicator of the availability of glucose in the luminal content of the small intestine. The presence of SGLT1 protein in the small intestine will support the hypothesis of rumen bypass and/or ruminal escape of nutrients. Furthermore we measured the maltase activities in the BBM, as part of the carbohydrates reach the intestinal lumen as polysaccharides and have to be broken down to monosaccharides before absorption.     

Influence of food deprivation on the transport of 3-O-methyl-alpha-D-glucose across the isolated ruminal epithelium of sheep

Recent studies provided evidence that the ruminal epithelium is able to absorb D-glucose even at physiologically low intraruminal concentrations. To elucidate whether ruminal D-glucose transport shows adaptive responses during food deprivation, transport of 3-0-methyl-alpha-D-glucose (3-OMG), a hardly metabolizable D-glucose analogue, was measured in isolated ruminal epithelia obtained from hay-fed or food-deprived adult sheep. In both groups, a significant net absorption of 3-OMG to the serosal side (in vivo: blood side oriented) could be detected at 3-OMG concentrations between 0.25 mM and 5 mM. Net absorption of 3-OMG was abolished by mucosal (in vivo: lumen side oriented) addition of phlorizin, an inhibitor of the sodium glucose-linked transporter 1 (SGLT-1). Net absorption of 3-OMG followed Michaelis-Menten kinetics, but apparent affinity and maximal transport capacity were lower in epithelia obtained from food-deprived sheep. In contrast to the decrease of the (secondary) active 3-OMG transport, serosal-to-mucosal permeation of 3-OMG increased after food deprivation, suggesting an elevated passive 3-OMG transfer. It is concluded that the altered transport characteristics are either part of a global energy-sparing process during food deprivation (i.e., a lowered activity of the Na+/K+-ATPase) or result from specific down-regulation of SGLT-1.     

Characterization of d-Glucose Transport across Equine Jejunal Brush Border Membrane Using the Pig as an Efficient Model of Jejunal Glucose Uptake

To test the hypotheses that glucose transport capacity across the brush border membrane (BBM) of the large colon is lower than that of the small intestine in equids, and that small intestinal transport capacity in equids is lower than suids. d-glucose transport capacity (V_max) and affinity (K_M) across the BBM of the distal jejunum (DJ) and proximal large colon (PLC) of the pony and pig were measured. Mucosa was collected from the DJ and PLC of ponies (n = 4) and pigs (n = 3), flash-frozen in liquid nitrogen, and stored at −80°C. Jejunal and colonic BBM vesicles were manufactured, and d-glucose transport was determined. There was no detectable active uptake of glucose in the equine PLC. Compared with the pig DJ, d-glucose transport capacity was lower (2595 ± 331 vs. 655 ± 286 ρmol·mg protein⁻¹·s⁻¹, respectively, P < .01) and transport affinity tended to be lower (0.09 ± 0.07 vs. 0.27 ± 0.06 mM, respectively, P = .11) in the pony DJ. Compared with the pig DJ, d-glucose transport capacity (2,595 ± 331 vs. 571 ± 331 ρmol·mg protein⁻¹·s⁻¹, respectively, P < .001) and transport affinity (0.09 ± 0.07 vs. 0.54 ± 0.07 mM, respectively, P < .001) in the pig PLC were lower. Results show there is negligible d-glucose uptake across pony PLC, and capacity for d-glucose absorption across DJ BBM is fourfold lower in the pony compared with the pig. Results further exemplify, at a physiological level, the limited capacity of the equid small intestine to transport glucose relative to an omnivorous mammal, and the likely evolutionary adaptation of equids to low dietary levels of nonstructural carbohydrates.     

肠道葡萄糖转运载体研究进展

D-葡萄糖是机体的主要能源物质,对机体代谢与内环境稳态有非常重要的作用。葡萄糖的吸收主要通过位于肠黏膜上皮细胞的两类葡萄糖转运载体家族来完成。Na+与SGLTs的结合促使载体与葡萄糖的结合,葡萄糖顺着Na+的浓度梯度进入细胞;当细胞内葡萄糖浓度升高后,葡萄糖顺着浓度差通过肠黏膜上皮细胞基底膜GLUT2经易化扩散转运进入血液。本文综述了肠道不同葡萄糖转运载体家族的成员和分类,介绍了其结构特征、功能特性及其组织分布;并详细阐述了肠道葡萄糖转运载体基因表达的影响因素。     

Lysine and methionine transport by bovine jejunal and ileal brush border membrane vesicles

Lysine (LYS) and methionine (MET) transport were studied using brush border membrane vesicles from bovine jejunal and ileal tissues. Total transport of LYS and MET was divided into mediated and diffusion components. Mediated uptake was further divided into sodium (Na)-dependent and Na-independent systems. Total LYS and MET uptake by ileal brush border (BB) vesicles tended to be higher than that by jejunal BB vesicles at all concentrations evaluated but differences were significant (P less than .05) at 2.5 and 7.5 mM for LYS and 5, 12.5 and 15 mM for MET. The greater capacity of ileal BB vesicles appeared to be due to the Na-dependent component of LYS uptake and the diffusion component of MET uptake. Transporters had less affinity but higher capacity than for LYS transport in both ileal and jejunal tissue. Methionine transport was greater (P less than .05) than LYS transport in both ileal and jejunal BB vesicles when the initial amino acid concentration was 7.5 mM. But when the initial amino acid concentration was 1.25 mM, MET uptake was greater (P less than .13) than LYS uptake in jejunal, but not in ileal, BB vesicles. The relative contribution of mediated and diffusion uptake systems to total MET and LYS uptake was found to be dependent on substrate concentration. Both intestinal site and substrate concentration influenced the contributions of Na-dependent, Na-independent and diffusion systems to total methionine and lysine uptake.     

Characterization of L-lysine transport across equine and porcine jejunal and colonic brush border membrane

In nonruminant herbivores, microbially derived AA could contribute to whole-body AA homeostasis and thus decrease predicted AA requirements. However, postileal capacity of AA uptake is currently unknown. Therefore, to test the hypothesis that Lys is transported across the large colon mucosal apical membrane with capacity similar to that of the small intestinal mucosa in the pony and pig, we examined Lys transport in vitro using brush border membrane vesicles (BBMV). Mucosa was collected from the distal jejunum (DJ) and proximal large colon (PLC) of growing pigs (n = 3) and ponies (n = 4), flash frozen in liquid nitrogen, and stored at -80°C. Jejunal and colonic BBMV were manufactured by Mg(2+) precipitation and used to determine initial rates and kinetics [the maximal transport rate (V(max)) and the Michaelis constant (K(M))] of l-Lys transport into apical epithelia by rapid filtration technique in Na(+)-gradient incubation buffer. Initial rates of total l-Lys uptake did not differ between the PLC and DJ in either the pig or the pony, or between the pony and the pig, at each l-Lys concentration. In the pig, compared with the DJ, l-Lys transport V(max) in the PLC did not differ (121 ± 26 and 180 ± 26 pmol?mg of protein(-1)?s(-1), respectively; P = 0.14) and l-Lys K(M) in the PLC tended to be greater (0.23 ± 0.22 and 0.89 ± 0.22 mM, respectively; P = 0.09). In the pony, compared with the DJ, l-Lys transport V(max) in the PLC was greater (62 ± 25 and 149 ± 25 pmol?mg of protein(-1)?s(-1), respectively; P = 0.04) and l-Lys K(M) in the PLC was greater (0.08 ± 0.22 and 1.05 ± 0.22 mM, respectively; P = 0.02). l-Lysine diffusion was not different between segments; however, total intestinal diffusion was greater (P = 0.03) in the pony than in the pig (115 ± 10 and 73 ± 10 pmol·mg of protein(-1)?s(-1), respectively). These results demonstrate that the large colon is capable of l-Lys transport across the apical epithelial membrane with greater capacity and less affinity than the jejunum, indicating that the large colon may play a significant role in l-Lys absorption and homeostasis in hindgut fermenters.     

Effects of epidermal growth factor on placental amino acids uptake in pregnant rats

To evaluate the effects of epidermal growth factor (EGF) on placental amino acids uptake, transport activities for L-proline, L-leucine, and L-alanine were kinetically examined in placental microvillous vesicles(PMV) obtained from pregnant rats administered with EGF(100 and 200 microg/kg/day) from day 18 to 21 of pregnancy. The Vmax of Na(+)-dependent proline uptake remarkably increased with a dose-dependent manner of EGF, while Km did not change. In contrast, Vmax and Km values of Na(+)-dependent and -independent alanine, and Na(+)-independent leucine uptake were not affected. These results suggested that EGF enhanced proline transport activity in placental microvillous membranes, resulting in an increase of proline concentration in the fetal blood. The selective up-regulation of proline uptake was considered to contribute to fetal growth by EGF.     

解淀粉芽孢杆菌复合菌剂对玉米秸秆的降解作用及表征

为获得能提高秸秆饲草品质的微生物发酵菌剂,本研究在前期获得2株具有木质素和纤维素降解能力的解淀粉芽孢杆菌菌株的基础上,将两者混配成复合菌剂,考察复合菌剂对玉米秸秆的降解作用,并通过傅立叶红外光谱(FTIR)、核磁共振氢谱(¹H-NMR)、扫描电镜(SEM)及气质联用色谱(GC/MS)等技术对玉米秸秆的微观结构及降解产物进行表征。研究发现,复合菌剂可以有效降解玉米秸秆中的木质纤维素。在发酵24 d时,玉米秸秆中木质素、纤维素和半纤维素的降解率分别达到48.4%,30.5%和41.4%。FTIR和¹H-NMR谱图中能观察到木质纤维素分子结构中主要连接共价键,如木质素单体间的β-O-4和β-β键、木质素与碳水化合物连接键以及碳水化合物中糖环内的价键等明显断裂,木质纤维素被部分降解;SEM扫描电镜图则显示发酵后秸秆的组织结构出现松散和破坏。发酵后秸秆中小分子物质的GC/MS分析结果显示,其中包含苯丙胺和苯丙酸等保留苯丙烷结构单元的木质素单体衍生物以及苄醇和苯甲酸酯类等木质素单体被进一步降解后的芳香族化合物。玉米秸秆中碳水化合物的GC/MS分析结果表明:复合菌剂可将玉米秸秆中的结构性多糖等大分子碳水化合物降解成葡萄糖、木糖、甘露糖及乳糖等还原性单糖。并利用这些还原性单糖生长代谢,进一步产生乙二醇、丙三醇及短链脂肪酸类等代谢产物。以上研究结果表明,解淀粉芽孢杆菌复合菌剂可有效降解玉米秸秆中的木质纤维素,在玉米秸秆饲草化利用中极具应用前景。     

Ruminant Nutrition Symposium: Role of fermentation acid absorption in the regulation of ruminal pH

Highly fermentable diets are rapidly converted to organic acids [i.e., short-chain fatty acids (SCFA) and lactic acid] within the rumen. The resulting release of protons can constitute a challenge to the ruminal ecosystem and animal health. Health disturbances, resulting from acidogenic diets, are classified as subacute and acute acidosis based on the degree of ruminal pH depression. Although increased acid production is a nutritionally desired effect of increased concentrate feeding, the accumulation of protons in the rumen is not. Consequently, mechanisms of proton removal and their quantitative importance are of major interest. Saliva buffers (i.e., bicarbonate, phosphate) have long been identified as important mechanisms for ruminal proton removal. An even larger proportion of protons appears to be removed from the rumen by SCFA absorption across the ruminal epithelium, making efficiency of SCFA absorption a key determinant for the individual susceptibility to subacute ruminal acidosis. Proceeding initially from a model of exclusively diffusional absorption of fermentation acids, several protein-dependent mechanisms have been discovered over the last 2 decades. Although the molecular identity of these proteins is mostly uncertain, apical acetate absorption is mediated, to a major degree, via acetate-bicarbonate exchange in addition to another nitrate-sensitive, bicarbonate-independent transport mechanism and lipophilic diffusion. Propionate and butyrate also show partially bicarbonate-dependent transport modes. Basolateral efflux of SCFA and their metabolites has to be mediated primarily by proteins and probably involves the monocarboxylate transporter (MCT1) and anion channels. Although the ruminal epithelium removes a large fraction of protons from the rumen, it also recycles protons to the rumen via apical sodium-proton exchanger, NHE. The latter is stimulated by ruminal SCFA absorption and salivary Na(+) secretion and protects epithelial integrity. Finally, SCFA absorption also accelerates urea transport into the rumen, which via ammonium recycling, may remove protons from rumen to the blood. Ammonium absorption into the blood is also stimulated by luminal SCFA. It is suggested that the interacting transport processes for SCFA, urea, and ammonia represent evolutionary adaptations of ruminants to actively coordinate energy fermentation, protein assimilation, and pH regulation in the rumen.     

妊娠后期营养限制对母羊胃肠道葡萄糖转运载体相关基因表达的影响

本研究旨在研究妊娠后期营养限制对母羊胃肠道葡萄糖转运载体相关基因表达的影响。选取20只同期受孕的湘东黑山羊,随机分为2组,即对照组(自由采食)和限饲组(40%采食量限制),每组10只。预试期15 d(妊娠81~95 d),正试期39 d(妊娠96~135 d)。正试期结束后,屠宰并采取瘤胃、十二指肠、空肠、回肠以及盲肠的黏膜样品,利用实时定量PCR技术,检测Na+-葡萄糖共转运载体1(SGLT1)、Na+-葡萄糖共转运载体3(SGLT3)、易化葡萄糖转运载体2(GLUT2)和易化葡萄糖转运载体5(GLUT5)基因表达量。结果表明:限饲组与对照组相比,SGLT1基因表达量在瘤胃显著降低(P0.05),在空肠和回肠中有降低趋势(0.05≤P0.10);GLUT5基因表达量在盲肠显著降低(P0.05);而其他葡萄糖转运载体基因胃肠道表达量在限饲组和对照组差异均不显著(P0.05)。由此可见,母羊妊娠后期营养限制对胃肠道中葡萄糖转运载体基因表达有不同程度的影响,进而引起母羊机体内葡萄糖转运的改变。