长途运输应激对孕中期小鼠胚胎发育的影响

为研究运输应激对胚胎发育的影响机制,采用普通公路运输建立小鼠运输应激模型(1 000 km,15 h)。将27只孕中期(怀孕8～10 d)的小鼠随机分为对照组(13只)和应激组(14只),分析运输应激对胎儿发育的影响;之后2组小鼠随机挑选各3只,采用agilent小鼠全基因组表达谱芯片筛选长途运输应激小鼠垂体和子宫中的差异表达基因。选取6个差异表达基因,应用实时荧光定量PCR技术验证芯片结果的可靠性。结果表明:与对照组相比,长途运输导致活胎率下降和死胎率增加(P<0.05);在孕中期垂体中筛选到138个基因差异表达,其中71个基因表达显著上调,67个基因表达显著下调;在孕中期子宫中筛选到392个基因差异表达,其中73个基因表达显著上调,319个基因表达显著下调。对差异显著基因进行IPA分析发现,主要通路和生物功能涉及免疫和炎性疾病、细胞的生长、增殖和细胞间的相互作用、机能失调等;实时荧光定量PCR对akap3、Angptl4、Cartpt、insl6、thy1进行验证结果与芯片结果的表达趋势一致。     

P‐glycoprotein in sheep liver and small intestine: gene expression and transport efflux activity

The role of the transporter P‐glycoprotein (P‐gp) in the disposition kinetics of different drugs therapeutically used in veterinary medicine has been demonstrated. Considering the anatomo‐physiological features of the ruminant species, the constitutive expression of P‐gp (ABCB1) along the sheep gastrointestinal tract was studied. Additionally, the effect of repeated dexamethasone (DEX) administrations on the ABCB1 gene expression in the liver and small intestine was also assessed. The ABCB1 mRNA expression was determined by real‐time quantitative PCR. P‐gp activity was evaluated in diffusion chambers to determine the efflux of rhodamine 123 (Rho 123) in the ileum from experimental sheep. The constitutive ABCB1 expression was 65‐fold higher in the liver than in the intestine (ileum). The highest ABCB1 mRNA expression along the small intestine was observed in the ileum (between 6‐ and 120‐fold higher). The treatment with DEX did not elicit a significant effect on the P‐gp gene expression levels in any of the investigated gastrointestinal tissues. Consistently, no significant differences were observed in the intestinal secretion of Rho 123, between untreated control (P_eff S‐M = 3.99 × 10^?6 ± 2.07 × 10^?6) and DEX‐treated animals (P_eff S‐M = 6.00 × 10^?6 ± 2.5 × 10^?6). The understanding of the efflux transporters expression and activity along the digestive tract may help to elucidate clinical implications emerging from drug interactions in livestock.     

LPS对肉鸡空肠多药耐药相关蛋白2(MRP2)mRNA表达及其底物酮康唑小肠渗透性的影响

为了探讨脂多糖对肉鸡空肠中多药耐药相关蛋白2(MRP2)mRNA表达及转运功能的影响,采用荧光定量PCR方法检测脂多糖处理6 h和12 h后对健康肉鸡空肠组织中MRP2 mRNA表达的影响;采用小肠原位单向灌流试验测定了不同剂量脂多糖(5 mg/kg和15 mg/kg)对酮康唑空肠渗透性的影响。结果显示:经LPS处理6和12 h后,空肠中MRP2 mRNA的表达水平均极显著降低(P<0.01);鸡小肠原位单向灌流试验显示高浓度的脂多糖(15 mg/kg)能显著增加酮康唑在小肠的有效渗透率(Peff)(P<0.05)。试验表明,脂多糖能抑制肉鸡空肠组织中MRP2 mRNA的表达并对空肠的外排转运功能产生影响,从而促进其底物药物的小肠渗透性,进而可能会影响其体内生物利用度。该结果可为炎症状态下肉鸡的合理用药提供理论支持。     

低温胁迫对狗牙根生理及基因表达的影响

以耐寒性差异较大的杂交狗牙根品种运动百慕大、天堂419和普通狗牙根品种保定狗牙根为试验材料,分析了昼夜温度为适温(30 ℃/25 ℃)、亚适温(18 ℃/12 ℃)、冷害(8 ℃/4 ℃)和冻害(4 ℃/-4 ℃)4种温度处理下,低温对狗牙根生长速率和草坪质量、MDA含量、叶绿素荧光(F_v/F_m)、光合作用(P_n、G_s、C_i、T_r)、H₂O₂和O2·-含量、抗氧化酶活性(SOD、POD、CAT、APX)、抗氧化酶及耐寒相关基因(CBF1、COR、LEA)表达的影响。结果表明,随着温度的降低,狗牙根草坪质量、生长速率、F_v/F_m和光合作用显著下降,但在耐寒性强的运动百慕大中下降幅度较小;低温下叶片MDA和H₂O₂含量及O2·-产生速率显著升高,其中在耐寒性弱的保定狗牙根中升高程度更大;狗牙根叶片抗氧化酶活性及抗氧化酶基因的表达水平随着处理温度的下降而显著升高,其中在运动百慕大中更为明显;狗牙根CBF1、COR和LEA基因表达量随着温度的下降而显著升高,尤其是在耐寒狗牙根运动百慕大中更为显著。低温诱导的抗氧化酶和耐寒相关基因上调表达,增强了细胞内的抗氧化防御,有助于狗牙根植株维持较高的光合作用和细胞膜稳定性,延缓叶片的衰老,从而提高了狗牙根的抗寒能力。     

低聚木糖对断奶仔猪肠道pH和肠黏膜形态结构的影响

本试验通过在日粮中添加低聚木糖探讨其对早期断奶仔猪肠道结构功能的影响。试验选用72头35日龄、体重9.81±0.41 kg杜＆#215;长＆#215;大三元杂交仔猪,随机分配成4组,每组3个重复,每个重复6头仔猪,公母各半。第1组为对照组,饲喂玉米-豆粕基础日粮;其余3组分别在各组日粮中添加0.01%、0.02%、0.04%低聚木糖作为试验组。试验预饲期2 d,正试期为28 d。于试验的最后1 d从各重复中随机选取1头仔猪屠宰,并测定胃、十二指肠、空肠、回肠、盲肠和结肠内容物的pH及十二指肠、空肠和回肠黏膜上皮的绒毛高度和隐窝深度。试验结果显示,试验组猪的胃及各肠段的pH比对照组均有所降低,尤其是0.02%低聚木糖组,差异显著（P＜0.05）;试验组各肠段的绒毛高度均高于对照组,差异显著（P＜0.05）,隐窝深度低于对照组,但差异不显著（P＞0.05）,绒毛高度与隐窝深度之比显著高于对照组（P＜0.05）。本试验结果表明,在早期断奶仔猪的日粮中添加低聚木糖,能降低胃肠道内容物的pH,改善早期断奶仔猪肠道的形态结构,以0.02%低聚木糖组效果最为明显。     

肉用犊牛产生应激的原因及机理分析

肉用犊牛在转至育肥场会出现一系列的应激反应,对肉牛育肥产业带来极大损失,本文通过对犊牛运输应激及断奶应激全面剖析的基础上,对产生犊牛应激综合征提出了新的看法,并提出要从犊牛的心理、断奶、环境改变、饲草料变化和运输等方面所产生的一系列应激叠加角度上考虑,综合预防犊牛过渡应激的横发,降低犊牛应激综合征给肉牛生产造成的损失。     

Extrusion decreases the negative effects of kidney bean on enzyme and transport activities of the rat small intestine

The objective of the present study was to evaluate the influence of raw and extruded kidney bean (Phaseolus vulgaris L. var. Pinto) consumption on the gut physiology of young growing rats. The intestinal enzyme activity (sucrase, maltase, Na(+) /K(+) ATPase, aminopeptidase N, dipeptidylpeptidase IV, alkaline phosphatase) and the uptake of sugar (d-galactose) and amino acids (l-leucine) were measured in brush border membrane vesicles. Five groups of growing male Wistar rats were fed ad libitum for 15 days on five different 10% protein diets: one containing casein as the main source of protein (Control, C), and four containing raw (RKB1, RKB6) or extruded kidney bean (EKB1, EKB6) at 1% and 6% of total protein content respectively. Extrusion treatment significantly reduced the content of bioactive factors (phytates, tannins) and abolished lectins, trypsin, chymotrypsin, and α-amylase inhibitory activities. Rats fed raw beans (especially RKB6) showed lower growth rate and food intake as compared to those fed extruded legumes, probably due to the high levels of lectins and other anti-nutritive factors in the raw beans. Gut enzymatic activities and uptake of d-galactose and l-leucine were lower in RKB6 and RKB1-fed animals, although they significantly improved in the groups fed extruded beans. Enzymatic activity and uptake in EKB1 were similar to those of casein-fed rats, whereas the uptake and growth rate of EKB6 were different to the control. This is attributable to the higher non-thermolabile biofactor content in the EKB6 diet, especially phytates and tannins, than in EKB1. This article shows the dose-dependent toxicological effects of bioactive factors contained in kidney beans on gut function. The extrusion process reduced their adverse impact on gut physiology and growth rate.     

寒冷应激对阿勒泰羊硬脂酰辅酶A去饱和酶表达的影响以及序列分析

本试验以阿勒泰羊为研究对象,对硬脂酰辅酶A去饱和酶(SCD)基因的部分编码区的cDNA进行克隆、测序,与GenBank中已收录的其他12种动物的SCD基因序列进行同源性分析,并构建分子进化树,采用RT-PCR方法,研究不同的寒冷应激温度对阿勒泰羊SCD mRNA表达的影响。结果显示,所克隆的SCD基因与绵羊SCD基因序列同源性最高,达98.91%;与虾夷扇贝和斑马鱼序列同源性最低,分别为56.52%与63.14%。在寒冷条件下阿勒泰羊和湖羊各种组织中SCD mRNA的表达都有所增加,阿勒泰羊尤为显著。结果表明,寒冷应激条件下,阿勒泰羊组织中表达较高水平的SCD,其高效表达能够提高脂膜的流动性,提升抵御低温的能力,这说明阿勒泰羊对寒冷应激的耐受性高于湖羊,体现了阿勒泰羊的种间优越性。序列分析结果也为阿勒泰羊SCD基因的生物学功能研究和遗传进化研究提供分子依据。     

Effect of acute deficiency of dietary proline on proline balance in the rat small intestine and liver

Introduction Proline is widely found in all types of mammalian tissue, and accounts for about 20% of the amino acids that constitute collagen (A dams 1970). Proline is nutritionally nonessential but biologically it is an important amino acid and consequently mammalian organisms synthesize the required amounts of proline even in the absence of sufficient proline consumption via food. The metabolism of higher animals is unique, and amino acid metabolism differs from one tissue to the next. Some organs are capable of synthesizing nonessential amino acids for use throughout the body. For example, arginine is primarily synthesized in the kidney and then released and distributed throughout the body. Pyrroline-5-carboxylate reductase is the enzyme responsible for the final stage of proline synthesis, and its activity has been confirmed in many important organs and tissue such as the cartilage, liver, small intestine, kidney and thymus gland (H erzfeld et al. 1977; S mith and P hang 1978). However, the different levels of pyrroline-5-carboxylate reductase activity among these organs has led to the belief that different amounts of proline are synthesized in these organs. In order to ascertain biological responses to dietary proline deficiency, it is important to identify the organs that release and distribute proline throughout the body when insufficient proline is consumed through the diet, thus reducing the blood proline concentration. Few studies have investigated this issue, but when ascertaining biological responses to dietary proline deficiency, it is more important to elucidate the effect of dietary proline deficiency on the metabolism of proline and other amino acids that are closely related metabolically to proline, in proline synthesizing organs. One of the most effective ways to assess amino acid metabolism in a target tissue of higher animals is to measure the difference between the arterial and venous concentrations of amino acids. I shikawa (1974) measured arteriovenous differences in order to examine the release of proline from the kidney and small intestine of fasted rats and the uptake of proline by the liver. In a previous study, it was found that when the plasma proline concentration was reduced to the fasting level by the consumption of a proline-deficient diet, proline was released from the kidney (W atanabe et al. 1995, 1997). In the present study, to ensure the induction of dietary proline deficiency, a completely purified diet containing all amino acids except for proline was prepared and fed to rats under experimental conditions. To investigate the role of the small intestine and liver in supplying and ingesting proline when the uptake of proline through food is restricted, the release and uptake of amino acids in the small intestine and liver were assessed by measuring carotid artery–portal vein and portal–hepatic vein differences in proline in rats.     

Effect of heat-stable enterotoxin of Escherichia coli and theophylline on ion transport in porcine small intestine.

The effect of a heat-stable enterotoxin of Escherichia coli was compared with that of theophylline on ion transport in the pig jejunum, using both in vivo and in vitro techniques. The maximal electrical response to heat-stable enterotoxin was only one-half that of theophylline even though the magnitude of the net secretory response was similar. A net, active secretion of HCO3 was partially responsible for the secretory response induced by heat-stable enterotoxin, whereas theophylline induced an active secretion of chlorine which could account for the entire secretory response. Heat-stable enterotoxin elevated tissue cyclic guanosine monophosphate levels, whereas theophylline elevated both cyclic adenosine monophosphate and cyclic guanosine monophosphate. Cyclic guanosine monophosphate levels induced by heat-stable enterotoxin were markedly potentiated by theophylline. Results suggest that HCO3 secretion in the pig jejunum may be controlled by the cyclic guanosine monophosphate system and this system also activates a neutral secretory process which at high heat-stable enterotoxin doses accounts for the bulk of the net secretion observed. Conversely, the chlorine secretion elicited by theophylline is entirely electrogenic and is consistent with results obtained in other species.     

Pre-weaning eating activity and morphological parameters in the small and large intestine of piglets

The effect of pre-weaning eating activity on the intestinal morphology in piglets was investigated on the day of weaning and 5 days post-weaning. The eating activity of each piglet was recorded by means of direct observations. On day 5 post-weaning the villous heights were reduced and the crypt depths were increased irrespective of the pre-weaning eating activity. In the caecum and colon the crypt depth was increased post-weaning and no effect of pre-weaning eating activity was observed. The muscle thickness in the small intestine did not change during the immediate post-weaning period whereas the muscle thickness in the large intestine was doubled in the same period. In conclusion, pre-weaning eating activity did not affect the structural alterations of the small and large intestine. However, the actual number of piglets consuming creep feed prior to weaning was low and the eating activity was highly variable. Therefore, more or later weaned piglets may be needed to show effects of pre-weaning eating activity.     

Special sugar expression on apoptotic epithelial cells of Peyer's patches and intestinal villi in rat small intestine

Our previous study clarified that the apical regions of both the follicle-associated epithelium (FAE) of Peyer's patches and the intestinal villi are the only adhesion sites of indigenous bacteria in rat jejuno-ileum. To survey the ligands against bacterial lectins, sugar expression patterns on epithelial cells were lectin-histochemically investigated using 21 lectins in the jejuno-ileal Peyer's patches of rats. As a result, (D-glcNAc)(2-4), detected by Solanum tuberosum (STL) and by Lycopersicon esculentum (LEL), and beta-D-gal(1-3)-D-galNAc detected by Peanut agglutinin (PNA), were strongly expressed on the brush borders of the apical regions of the FAE and the intestinal villi. On the other hand, neither sugar was expressed on the brush borders of the basal regions of both FAE and intestinal villi. The positive intensities for the lectins correlated with the progression of epithelial apoptosis in the FAE and in the intestinal villi. Moreover, the double staining with lectin histochemical method and the in situ nick end-labeling method could simultaneously detect the strong expression of both sugars and nuclear DNA fragmentation in epithelial cells at the late apoptotic stage. Other sugar expression patterns in the intestinal villi were similar with those in the FAE. There were no lectins specific for M cells in the FAE. From these findings, the possible sugars of ligands against some indigenous bacterial lectins, expressing specially on the apoptotic epithelial cells, might be narrowed down in rat jejuno-ileum.     

Low crude protein diets supplemented with casein hydrolysate enhance the intestinal barrier function and decrease the pro-inflammatory cytokine expression in the small intestine of pigs

To reduce nitrogen excretion and lower feeding costs, low crude protein (CP) diets are sometimes proposed, however, a great reduction of dietary CP concentration (>4% reduction vs. recommended concentration), even supplemented with essential and nonessential amino acids (AA) can detrimentally affect small intestinal barrier function and immunity, possibly due to the excessive lack of peptides. Here we hypothesize that with an extremely low CP concentration diet, protein-derived peptides, rather than AA supplementation, can improve intestinal barrier development and health. To test this hypothesis, 21 growing pigs (19.90 ± 1.00 kg body weight) were randomly assigned to 3 treatments with control diet (16% CP), or low CP diets (13% CP) supplemented with AA (LCPA) or casein hydrolysate (LCPC) for 28 days. In comparison with the control diet, the LCPA diet decreased the protein expression level of jejunal barrier factor zonula occludens-1 (ZO-1) and stem cell proliferation factor leucine-rich repeat-containing G-protein-coupled receptor-5, whereas the LCPC diet enhanced intestinal barrier function by increasing the protein expression level of jejunal occludin and ZO-1 and ileal mucin-2. The LCPA diet reduced Lactobacillus counts, whereas the LCPC diet increased Lactobacillus counts and reduced Escherichia coli counts in the ileum. The LCPA diet also increased protein expression levels of pro-inflammatory cytokine interleukin-6 (IL-6) and IL-22, whereas the LCPC diet decreased protein expression levels of pro-inflammatory IL-1β, IL-17A and tumor necrosis factor-α in the ileum. Collectively, the casein hydrolysate supplementation of low CP diets showed beneficial effects on the small intestinal barrier, bacterial community, and immunity in pigs, pointing to the important role of protein-derived peptides in small intestinal health in cases of low crude protein diets.     

运输应激对2种Hal基因型猪肌纤维特性的影响

以二花脸与皮特兰杂交F2代猪为研究对象,应用组织学、组织化学方法与电镜技术研究运输应激对2种猪背最长肌琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、糖原以及肌纤维超微结构的影响,并分析比较了2种氟烷基因(HalNN,Halnn)猪背最长肌各肌纤维类型。研究发现:(1)HalNN猪背最长肌SDH活性比Halnn猪的极显著高(P<0.01);(2)背最长肌LDH活性Halnn猪比HalNN猪的极显著高(P<0.01);(3)Halnn猪糖原含量比HalNN猪的显著高(P<0.05);(4)透射电镜显示HalNN猪背最长肌肌纤维超微结构形态较清晰完整,而Halnn猪肌纤维收缩剧烈,肌节混乱,明带暗带区分不明显,有的肌节甚至降解,线粒体不同程度肿胀和裂解,肌浆网不明显等。结果表明:氟烷基因野生型HalNN猪对运输应激相对不敏感;而阴性纯合子Halnn猪则对应激很敏感,其肌纤维损伤较大。     

Ultrastructural study on the morphological changes in indigenous bacteria of mucous layer and chyme throughout the rat intestine

Indigenous bacteria in the alimentary tract are exposed to various bactericidal peptides and digestive enzymes, but the viability status and morphological changes of indigenous bacteria are unclear. Therefore, the present study aimed to ultrastructurally clarify the degeneration and viability status of indigenous bacteria in the rat intestine. The majority of indigenous bacteria in the ileal mucous layer possessed intact cytoplasm, but the cytoplasm of a few bacteria contained vacuoles. The vacuoles were more frequently found in bacteria of ileal chyme than in those of ileal mucous layer and were found in a large majority of bacteria in both the mucous layer and chyme throughout the large intestine. In the dividing bacteria of the mucous layer and chyme throughout the intestine, the ratio of area occupied by vacuoles was almost always less than 10%. Lysis or detachment of the cell wall in the indigenous bacteria was more frequently found in the large intestine than in the ileum, whereas bacterial remnants, such as cell walls, were distributed almost evenly throughout the intestine. In an experimental control of long-time-cultured Staphylococcus epidermidis on agar, similar vacuoles were also found, but cell-wall degeneration was never observed. From these findings, indigenous bacteria in the mucous layer were ultrastructurally confirmed to be the source of indigenous bacteria in the chyme. Furthermore, the results suggested that indigenous bacteria were more severely degenerated toward the large intestine and were probably degraded in the intestine.     

外源 Si 对盐胁迫下甘草幼苗形态及生理指标的影响

为深入了解 Si 对盐胁迫下甘草幼苗的影响,本实验采用培养皿滤纸床进行发芽试验,以乌拉尔甘草幼苗为试验材料,在两种浓度 NaCl 溶液胁迫下设置不同浓度的硅溶液对甘草幼苗进行处理,通过测定幼苗形态（胚根、胚芽）及生理（质膜相对透性、丙二醛和过氧化物酶）指标,以探讨在盐胁迫下外源硅对甘草幼苗形态及生理指标的影响。结果表明,在 NaCl 胁迫下,加入硅（Si）溶液后可以有效增加幼苗胚芽和胚根的长度及重量。在低盐（100 mmol／L）胁迫下,Si 浓度为0．4 mmol／L 时对胚根生长的促进最强;在高盐（200 mmol／L）胁迫下,Si 浓度为0．8 mmol／L 时对胚根生长的促进作用最强。外源 Si 也不同程度提高了过氧化物酶活性、渗透调节物质脯氨酸含量,以及丙二醛含量和质膜相对透性。由此可见,在盐胁迫条件下,加入适宜浓度的外源硅,可以通过保持一定的抗渗透胁迫能力和清除活性氧的能力,进而保持膜的稳定,来缓解盐胁迫对甘草幼苗生长的抑制作用。     

模拟干旱胁迫下镁对高羊茅种子萌发的影响

采用PEG-6000(100,200,300g/L)模拟水分胁迫,研究不同浓度的硫酸镁溶液(0.2,0.4,0.6,0.8g/L)浸种对高羊茅(Festuca arundinacea)种子萌发的影响。结果表明:随着干旱胁迫程度的增加,高羊茅的发芽率,发芽指数,苗长,根长,幼苗鲜重及根鲜重等指标均呈下降趋势。在种子萌发阶段,轻度干旱胁迫(PEG=100g/L)下,硫酸镁浸种对高羊茅种子的萌发促进作用不明显,其发芽率、发芽势、发芽指数与对照差异不显著;中度干旱胁迫(PEG=200g/L)下,0.6g/L的镁使得高羊茅种子的萌发相对最佳,其发芽等指标与对照差异显著(P0.05);重度干旱胁迫下(PEG=300g/L),0.4g/L镁处理明显的促进了高羊茅种子的萌发,其发芽指标与对照差异显著(P0.05),而当镁浓度是0.8g/L时,则抑制了高羊茅种子的萌发。在幼苗的生长前期,轻度干旱和中度干旱胁迫下,0.6g/L镁处理对高羊茅的苗长、根长、幼苗鲜重及根鲜重等生长指标有显著的促进作用;重度干旱胁迫下,0.4g/L和0.6g/L的镁处理使其生长指标高于对照且差异显著(P0.05)。