牦牛肠源Lactobacillus acidophilus L3对其小肠黏膜形态结构的影响

为了探讨牦牛肠源嗜酸乳杆菌Lactobacillus acidophilus L3对牦牛小肠肠道黏膜形态结构的影响,将10头健康牦牛(2~2.5岁)随机分为2组,分别为益生菌组和空白对照组,益生菌组在饲料中添加2×109CFU·kg-1L.acidophilus L3,饲喂28 d后取其小肠组织样品,采用组织学和显微测量学方法对益生菌组和空白对照组牦牛小肠绒毛高度、隐窝深度、绒毛高度/隐窝深度比值(V/C),以及回肠黏膜淋巴集结厚度进行测量和统计分析。研究结果显示,益生菌组牦牛十二指肠、空肠、回肠绒毛高度均较空白对照组显著增加,隐窝深度降低,V/C值增加,差异有统计学意义(P<0.05)。通过对回肠淋巴集结的测量发现,益生菌组牦牛回肠淋巴集结的平均厚度均较空白对照组显著增加,差异有统计学意义(P<0.05)。研究结果表明,牦牛肠源L.acidophilus L3可有效改善其肠道黏膜结构,增强肠道黏膜免疫效应部位的免疫功能。     

小鼠肠道组织结构在低温环境下的适应性调整

对长期(3个月)处于低温条件下的小鼠各肠道组织结构的适应性变化进行了比较研究。低温驯化后小鼠的体重(P0.05)、消化道总长度(P0.05)、小肠长度(P0.01)和盲肠长度(P0.05)显著增加,而十二指肠肠腔隙截面积未发生显著变化,提示小鼠的肠腔隙体积在低温环境下明显增加。同时,低温驯化小鼠十二指肠的肠道和肠壁组织截面积均显著降低(P0.05),而其绒毛高则显著增加(P0.001),提示其黏膜层厚度增加,而黏膜下层结构趋于萎缩,表明小鼠肠道组织的形态结构为应对低温条件已经产生了适应性调整,即通过增加消化道的长度和营养物质的吸收面积来增加食物摄取量、食物吸收速率和吸收效率等,进而满足低温条件下小鼠能量需求的增加。相比之下,低温鼠的胃大小、盲肠重量、大肠长度与重量、直肠组织结构均未产生显著变化,暗示这些肠道结构对外界温度变化的敏感性相对较低。研究结果表明,小型哺乳类主要通过增大消化道长度和吸收面积来提高其消化效率,通过改变消化道的形态和结构来提高消化和吸收效率,以便更好地去适应环境温度变化而导致的能量需求变化。     

青海沙蜥消化道组织结构及嗜银细胞研究

为揭示青海沙蜥消化系统的组织结构,探索其高海拔适应的组织学基础,应用解剖学与石蜡切片、H.E染色和Grimelius银染法对青海沙蜥消化道组织结构和嗜银细胞进行研究。结果显示：青海沙蜥的消化道管壁结构分为4层,从内到外依次是粘膜层、粘膜下层、肌层和浆膜。消化道各段的长度和壁厚均存在显著差异,其中小肠最长,胃幽门部的管壁最厚。粘膜皱襞和绒毛的分布也存在差异,空肠部位的小肠绒毛数量最多,其次是十二指肠和回肠。嗜银细胞形状多样,广泛分布在消化道各段的黏膜上皮基部和黏膜上皮之间。胃体是嗜银细胞分布密度最高的部位,其次是贲门,回肠最低。与栖息在低海拔的有鳞类相比,青海沙蜥为适应高海拔环境,小肠的相对长度变长,胃体部嗜银细胞增多。     

体外法探究猪空肠和回肠黏膜微生物对低聚半乳糖和低聚甘露糖的发酵特性

【背景】小肠黏膜微生物是肠道菌群的重要组成部分,大量研究表明日粮添加低聚半乳糖（galacto-oligosaccharides,GOS）和低聚甘露糖（manno-oligosaccharides,MOS）能够调控猪的大肠菌群结构,但关于其调控小肠黏膜微生物的研究较少。【目的】通过体外发酵法探究猪空肠黏膜和回肠黏膜微生物发酵GOS和MOS的规律。【方法】以生长猪的空肠黏膜微生物和回肠黏膜微生物作为接种物,以GOS和MOS作为底物进行厌氧发酵,在发酵0、6、12、24 h时采样测定总菌数量、pH、氨态氮（ammonia nitrogen,NH₃-N）、菌体蛋白（microbial crude protein,MCP）和有机酸,在24 h收集微生物提取DNA进行细菌定量分析。【结果】在24 h时,回肠黏膜组的NH₃-N浓度显著低于空肠黏膜组,而MCP浓度显著高于空肠黏膜组（P<0.05）。在发酵的前6 h各组pH无明显变化,有机酸积累较少。在12 h时,MOS组的乳酸、乙酸、丁酸和总短链脂肪酸产量显著高于GOS组（P<0.05）,此时只有回肠黏膜组有少量丙酸产生。在24 h时,MOS回肠黏膜组乳酸产量最高而pH值最低（P<0.05）。相较于MOS组,GOS组显著提高了丙酸的产量（P<0.05）。相较于GOS组,MOS组显著提高了乙酸的产量,在空肠黏膜组中显著提高了丁酸和总短链脂肪酸的产量（P<0.05）。定量结果表明,在24 h时,各处理组的厚壁菌门数量都接近总菌数量,属于优势菌门。相较于MOS组,GOS组显著提高了拟杆菌门、链球菌属、韦荣氏球菌属和普拉梭菌细菌的数量,提高了空肠黏膜组中Clostridium cluster IV和回肠黏膜组中Clostridium cluster XIVa的数量（P<0.05）。相较于GOS组,MOS组显著提高了大肠杆菌和乳酸杆菌属的数量,提高了回肠黏膜组中罗氏菌属的数量（P<0.05）。【结论】猪小肠黏膜微生物对GOS和MOS具有不同的发酵模式,主要表现在有机酸的产生和促进细菌的增殖方面。GOS具有产丙酸优势,提高了拟杆菌门和韦荣氏球菌属的数量;MOS促进了乙酸的产生,提高了大肠杆菌和乳酸杆菌的数量。     

光周期和温度对布氏田鼠产热的影响

布氏田鼠（Ｍｉｃｒｏｔｕｓ ｂｒａｎｄｔｉ）分组驯化在：（１）长光照温暖环境（ＬＷ，１６Ｌ：８Ｄ，２５℃）；（２）长光照 低温环境（ＬＣ，１６Ｌ：８Ｄ，５℃）；（３）短光照温暖环境（ＳＷ，８Ｌ：１６Ｄ，２５℃）；（４）短光照低温环境（ＳＣ， ８Ｌ：１６Ｄ，５℃）。驯化四周后，长光照动物的体重比短光动物的体重有增加的趋势；四组动物的体温 没有明显差异；低温和短光照均促使静止代谢率（ＲＭＲ）增加；ＳＣ组动物的非颤抖性产热（ＮＳＴ）高 于ＬＷ组。低温诱导肝和褐色脂肪组织（ＢＡＴ）细胞线粒体蛋白增加，短光照再度地增强此作用。短 光照诱导肝细胞线粒体状态－４及状态－３呼吸活力增加，低温没有明显作用。低温和短光照均刺激肝 和ＢＡＴ线粒体细胞色素Ｃ氧化酶活力提高，但后者作用强度大于前者。低温明显激活ＢＡＴ线粒体 的ａ－磷酸甘油氧化酶的活力，短光照无明显影响、结果表明：低温和短光照均能提高布氏田鼠的产 热能力，短光照与低温因子两者的协同作用增强了对布氏四鼠热能代谢的调节。     

猪胃肠道黏膜二糖酶的性质

研究了猪小肠中麦芽糖酶,蔗糖酶和乳糖酶3种二糖酶的生化性质及活性分布。试验以3头“杜加”生长猪为对象,屠宰并刮取胃底部,十二指肠,空肠上段和回肠黏膜。其中空肠黏膜用于3种二糖酶生化性质的研究,包括酶的最适温度,热稳定性,最适pH ,pH稳定范围和金属离子对酶活性的影响;胃底部,十二指肠,空肠上段和回肠黏膜用于测定3种二糖酶活性以揭示其在胃扬中的分布规律。试验结果表明：麦芽糖酶,蔗糖酶和乳糖酶的最适反应温度分别为50,45和55℃,最适pH分别为6.8,6.5和6．0;乳糖酶的耐热温度（70℃）高于麦芽糖酶和蔗糖酶（50℃）;不同pH对3种二糖酶活性影响不大;金属离子Cu^2 和Fe^2 对3种二糖酶均有激活作用;而Mn^2 有抑制作用。此外,Zn^2 能抑制麦芽糖酶活性,提高蔗糖酶活性,而不影响乳糖酶活性。3种二糖酶活性在肠道中由高到低的分布为：空肠,回肠,十二指肠和胃;其中麦芽糖酶在空肠和回肠中活性相近。回肠和空肠黏膜中的麦芽糖酶活性均显著高于十二指肠和胃中的酶活性,十二指肠酶活性显著高于胃;空肠中蔗糖酶和乳糖酶活性显著高于回肠,十二指肠和胃底部。从3种二糖酶活性大小看,胃底部和十二指肠中的麦芽糖酶活性显著高于蔗糖酶和乳糖酶;空肠上段和回肠中的麦芽糖酶活性显著高于蔗糖酶活性,蔗糖酶活性又显著高于乳糖酶。上述结果表明,3种二糖酶的生化性质具有一定的差异,在胃肠道中的活性分布规律相似。     

GLP-2对小鼠小肠黏膜的作用及不同脏器分布的影响

目的：观察胰高血糖素样肽-2对小鼠小肠黏膜的作用效果及其受体在不同脏器的分布和表达。方法：选用5-6周龄的雄性健康BALB/C小鼠,体重17～20g。随机分为3组。对照组8只;脑内注射GLP-2组9只,每天按25μg/kg脑内注射GLP-2液2次,连续3天;腹腔注射GLP-2组9只,每天按250μg/kg腹腔注射GLP-2液2次,连续3天。3天后,处死小鼠,做组织切片进行HE染色,观察小鼠小肠黏膜的组织学变化,用免疫组化方法检测GLP-2R在不同脏器的表达和分布。结果：小鼠经腹腔注射GLP-2后,小肠不同肠段黏膜的绒毛高度较对照组明显增加（P〈0.05）,脑内注射组的肠黏膜无明显形态学变化;GLP-2R在小鼠胃、空肠与结肠部位均有表达,而食管与肝显阴性;腹腔注射组的GLP-2R表达较对照组显著下调（P〈0.05）。结论：GLP-2能刺激小肠黏膜上皮增厚,增加小肠不同肠段黏膜的绒毛高度;小鼠的胃、空肠与结肠经GLP-2（腹腔注射）处理后其受体表达下调。     

布氏田鼠对低温的适应性产热

将布氏田鼠置１到３０天的低温（６±１℃）环境下暴露,其体重与体温没有明显变化;静止代谢率（ＲＭＲ）、非震颤性产热（ＮＳＴ）及血清中三碘甲腺原氨酸（Ｔ３）含量,随冷暴露时间的延长而增加;肝线粒体状态３呼吸及细胞色素Ｃ氧化酶活力亦随冷暴露时间的延长而升高。肝线粒体蛋白质含量经冷暴露３０天时有明显升高,表明动物在低温适应中肝线粒体蛋白合成增加与呼吸功能增强是个体ＲＭＲ提高的细胞学机制之一。布氏田鼠褐色脂     

布氏田鼠适应性产热过程中血清甲状腺激素的变化

早春季节对布氏田鼠（Microtusbrandti）施以不同光照和温度处理,其血清甲状腺素（T4）含量没有明显变化;低温明显地升高了三碘甲腺原氨酸（T3）的含量和T3／T4的比率。短光照只在低温环境中能升高T3／T4的比率。秋节温暖环境中的短光照或褪黑激素（Melatonin,MLT）处理,都能升高血清T3的浓度和增加T3／T4的比率。表明甲状腺激素在布氏田鼠短光照和低温的适应性产热中起着重要的调控作用,褪黑激素诱导布氏田鼠产热与甲状腺激素有密切的联系。     

牦牛肠源Lactobacillus acidophilus L3对其肠道分泌型免疫球蛋白A及免疫相关因子的影响

为了研究牦牛肠源Lactobacillus acidophilus L3(嗜酸乳杆菌)对其肠道分泌型免疫球蛋白A(SIg A)及免疫相关因子的影响,将10头健康牦牛(2~2.5岁)随机分为2组,分别为益生菌组和空白对照组,益生菌组在饲料中添加2×109CFU·kg-1L.acidophilus L3,饲喂28 d后取其小肠样品,ELISA检测试验组和对照组十二指肠、空肠、回肠中SIg A、白细胞介素-2(IL-2)、IL-4、IL-6、干扰素γ(IFN-γ)的含量。研究结果表明,L.acidophilus L3可有效增加试验组牦牛肠道SIg A的分泌量(P<0.05),组内比较发现SIg A在回肠含量最高,其次为空肠和十二指肠。IL-2、IL-4、IL-6在试验组小肠中的表达量较对照组显著增加(P<0.05),试验组小肠IFN-γ的表达量较对照组降低(P<0.05)。证实L.acidophilus L3可提高牦牛肠道的黏膜免疫功能。     

Adaptation of intestinal morphology in the temperature-acclimated carp,Cyprinus carpio L.

Summary The effects of temperature and photoperiod acclimation upon the morphology of carp intestinal mucosa have been studied using morphometric techniques. Carp intestine showed an absence of anatomical regionalisation. There was a gradual reduction in the dimensions of villi along the tract. The decrease in the dimensions of the villi was greatest in the anterior half. Temperature acclimation had no effect on intestinal-somatic indices. Acclimation to 10° C or 30° C resulted in large differences in the dimensions of villi. Cold acclimation produced significant increases in mean villus height and breadth along the entire intestine. These villus shape changes resulted in a 58% increase in total mucosal surface area and a 102% increase in total volume of villi in cold-acclimated fish relative to warm-acclimated fish. Surface area of the unmodified intestinal tube increased with cold acclimation by 28%. The total number of villi remained unchanged by thermal acclimation. Because normalisation to a nominal surface area does not take account of the possibility of differentially developed mucosal surfaces in differently acclimated animals, experiments comparing transepithelial transport rates of differently-acclimated fish, using unstripped preparations, overestimates the differences in area-specific transport capacity.     

急性冷胁迫对中华鳖幼鳖肠道黏膜组织的影响

为了探究急性冷胁迫对中华鳖(Pelodiscus sinensis)幼鳖肠道不同区段黏膜组织学特征的影响, 实验检测了急性冷胁迫前后血清二胺氧化酶(Diamine oxidase, DAO)的活性, 同时观察了急性冷胁迫前后中华鳖肠道黏膜的相关组织形态的差异。DAO活性显示: (1)在第一次急性冷胁迫实验中, 中华鳖血清DAO活性随急性冷胁迫时间的增加而呈现降低趋势, 并在冷胁迫到达48h降到了最低水平; (2)在急性冷胁迫及复温实验中, 中华鳖血清DAO的活性, 在冷胁迫3d后显著降低, 但随着温度的恢复, DAO活性又恢复到正常水平。组织病理结果显示: (1)急性冷胁迫对中华鳖肠道(回肠后段和大肠)黏膜上皮的形态没有明显影响; (2)急性冷胁迫对回肠后段的杯状细胞数目、肠绒毛长度和绒毛长度/隐窝深度的比值没有显著影响, 但会使回肠后段黏膜厚度显著降低; (3)急性冷胁迫会使大肠的杯状细胞数目降低。这表明急性冷胁迫会改变中华鳖肠道黏膜的结构, 但在不同的肠段, 这种改变是不同的。回肠后段和大肠在同样的冷胁迫方式下黏膜机械屏障的不同变化情况, 提示中华鳖肠道各段对急性冷胁迫具有特殊的应对方式。     

Effects of Trichinella spiralis infection on intestinal pathology in mice lacking interleukin-4 (IL-4) or intestinal trefoil factor (ITF/TFF3)

The nematode Trichinella spiralis induces pathological changes in the small intestine of the host, which are known to be controlled by immune and inflammatory mediators. The detail of this control has still to be completely understood. Mice deficient in interleukin 4 (IL-4) or in intestinal trefoil factor/trefoil family factor 3 (ITF/TFF3) were infected with T. spiralis and the resultant changes in the intestinal mucosa followed by quantifying numbers of mucosal mast cells, goblet cells, Paneth cells and by monitoring structural changes in villus length and crypt depth. Mice lacking IL-4 were unable to mount a normal protective response to infection, such that worm survival was increased. These mice failed to mount a mucosal mast cell response, but did make goblet cell and Paneth cell responses comparable to normal controls. Mice lacking ITF/TFF3 similarly made normal levels of goblet cell and Paneth cell responses. They also underwent profound changes in mucosal architecture, with marked villus atrophy and crypt hyperplasia. These results are discussed in relation to known patterns of T cell and cytokine control of protective immunity to T. spiralis. They suggest that increased numbers of goblet cell and Paneth cell are not, by themselves, required for protective immunity. ITF/TFF3 appears not to influence cellular responses and does not alter parasite-induced pathological changes in the small intestine.     

Acute distal intestinal obstruction in gnotobiotic rats. Intestinal morphology and cell renewal.

1. Complete mechanical obstruction of the distal small intestine was produced in gnotobiotic rats. 72 h after the operation small intestinal morphology and epithelial cell renewal were investigated proximal and distal to the site of obstruction. 2. Proximal to the site of obstruction there were minor changes in villus height, base length and in villus cell number, a large increase in depth and diameter of the crypts and an approximately threefold increase in cell renewal. 3. Distal to the site of obstruction there were no differences between the intestines of rats with obstruction and controls. 4. The apparent lack of secretion by the goblet cells and the reduced number of intraepithelial leucocytes suggest that the barrier function of the small intestine is impaired in obstruction.     

The mucosa of the small intestine: development of the cellular lipid composition during enterocyte differentiation and postnatal maturation

Alterations in lipids linked to intestinal maturation and enterocyte differentiation were reviewed. The 3 main lipid components of cell membranes, ie cholesterol, phospholipids and glycolipids, were examined. Cell phospholipid content increases from the crypts to the mid-villus, which accounts for membrane development and organelle growth in differentiating cells. Changes in the proportion of phospholipid polar head groups occur in brush border membrane during postnatal maturation of the small intestine. The possibility that phospholipid fatty acid composition in differentiating cells might be altered by dietary lipids is discussed. Cholesterol biosynthesis mainly occurs in crypt and lower villus cells whereas its absorption from luminal content and esterification into lipoproteins occur in upper villus mature cells. Cholesterol cell content increases in mature cells in comparison to immature cells on the one hand, and in the distal by comparison with proximal parts of the intestine on the other. Increasing cholesterol content is generally correlated with decreasing membrane fluidity, which in turn could modulate functional properties of the mucosa. Glycosphingolipids are mainly found in the brush border membrane, which contains 20-30% glycolipids by weight of total lipids. These components tend to reinforce the membrane stability and significantly contribute to the surface properties of epithelial cells. The latter undergo noticeable changes during cell differentiation and postnatal maturation. Significant changes in both the glycosidic and lipophilic parts of glycosphingolipid molecules occur in differentiating cells and are of possible importance in the process of mucosal maturation. It is possible that the addition of a terminal sialic acid (sialyltransferase activity) instead of a terminal galactose (galactosyltransferase) to an endogenous acceptor (lactosylceramide) could constitute an important event in the differentiation process, and may account for the increasing content of hematosides along the intestinal villus of rat. Alterations in lipid counterpart mainly consist of hydroxylation of fatty acids in hematosides during postnatal maturation or in glucosylceramides during cell differentiation. Collectively these intestinal lipid changes may contribute in part to the development of mucosal barrier, selective permeability and functional properties of the mature intestinal mucosa.     

Intrauterine Growth Restriction Impairs Small Intestinal Mucosal Immunity in Neonatal Piglets

Intrauterine growth restriction (IUGR) is a very common problem in both piglet and human neonate populations. We hypothesized that IUGR neonates have impaired intestinal mucosal immunity from birth. Using neonatal piglets as IUGR models, immune organ weights, the weight and length of the small intestine (SI), intestinal morphology, intraepithelial immune cell numbers, levels of cytokines and immunoglobulins, and the relative gene expression of cytokines in the SI were investigated. IUGR neonatal piglets were observed to have lower absolute immune organ weight and SI length, decreased relative weights of the thymus, spleen, mesenteric lymph node, and thinner but longer SIs. Damaged and jagged villi, shorter microvilli, presence of autophagosomes, swelled mitochondria, and decreased villus surface areas were also found in the SIs of IUGR neonatal piglets. We also found a smaller number of epithelial goblet cells and lymphocytes in the SIs of IUGR neonates. In addition, we detected reduced levels of the cytokines TNF-α and IFN-γ and decreased gene expression of cytokines in IUGR neonates. In conclusion, IUGR was shown to impair the mucosal immunity of the SI in neonatal piglets, and the ileum was the major site of impairment.     

Pre- and postnatal development of differentiated functions in rat intestinal epithelial cells

A panel of monoclonal antibodies to intestinal cell surface components has been used to compare the expression of differentiation-specific antigens in the epithelial cells of fetal, suckling, and adult rat small intestine. Indirect immunofluorescence staining, and immunopurification of detergent-solubilized membrane proteins, followed by single- and two-dimensional slab gel electrophoretic analysis, have demonstrated that fetal intestinal cells (at day 21 of gestation) express most differentiation-specific markers typical of adult absorptive villus cells. A marked heterogeneity in antigen expression was observed among different villus cell populations in suckling rat intestine, and three cell surface components were identified which are exclusively present during this period of intestinal development. Striking changes in the patterns of antigen expression in crypt and villus cells, and variations in the apparent isoelectric points for most luminal membrane components, were associated with the maturation of the intestinal mucosa at weaning. These changes could not be prematurely induced by cortisone injection in newborn rats, suggesting that factors other than glucocorticoids are responsible for the postnatal development of the intestinal epithelium. These results suggest that basic differences in biological properties and regulatory mechanisms exist among intestinal epithelial cells at different stages of pre- and postnatal maturation.     

Effects of environmental temperature and exercise on the biochemical characteristics of hamster intestine

The effects of increasing environmental temperature and of exercise on some biochemical characteristics of the intestinal mucosa were analyzed in hamsters to determine whether damage occurs to the intestine during exercise, because long-distance runners complain of cramp, diarrhea, or retrostaltic symptoms, especially when exercise is performed at high temperatures. Two sets of experiments were carried out on groups of five animals. First, one group stayed at rest at 20 degrees C while another group performed exercise for 30 min at the same temperature. Second, one group of animals remained at rest at 20 degrees C for 16 h, a second group was placed at 32 degrees C for the same period, and a third group was subjected to the latter treatment but in addition performed two 20-min exercises. The animals were killed immediately after the experiment. After the small bowel was removed, biopsies were taken for histological examination, and the remaining small bowel tissue was homogenized for biochemical analysis. During exercise performed at 20 degrees C or during exposure to 32 degrees C, the DNA weight (expressed as a function of the protein weight) increased; the specific activity of sucrase, leucine aminopeptidase, diamine oxidase, and maltase decreased; spermine and putrescine content generally decreased; and the weight of mucosal proteins per length of intestine did not vary significantly. When exercise was performed at 32 degrees C, we noted few modifications in the values of the intestinal parameters tested, i.e., changes in only the weight of mucosa expressed as a function of bowel length and, perhaps, the spermine or putrescine content.     

Intectin, a novel small intestine-specific glycosylphosphatidylinositol-anchored protein, accelerates apoptosis of intestinal epithelial cells

Intestinal epithelial cells undergo rapid turnover and exfoliation especially at the villus tips. This process is modulated by various nutrients especially fat. Apoptosis is one of the important regulatory mechanisms of this turnover. Therefore, identification of the factors that control epithelial cell apoptosis should help us understand the mechanism of intestinal mucosal turnover. Here, we report the identification of a novel small intestine-specific member of the Ly-6 family, intectin, by signal sequence trap method. Intectin mRNA expression was exclusively identified in the intestine and localized at the villus tips of intestinal mucosa, which is known to undergo apoptosis. Intectin mRNA expression was modulated by nutrition. Intestinal epithelial cells expressing intectin were more sensitive to palmitate-induced apoptosis, compared with control intestinal epithelial cells, and such effect was accompanied by increased activity of caspase-3. Intectin expression also reduced cell-cell adhesion of intestinal epithelial cells.