抗生素所致腹泻大鼠肠屏障功能损伤及乳酸杆菌保护机制的研究

目的探讨抗生素对所致腹泻大鼠肠道屏障功能、肠道菌群结构和肠道细菌移位的影响及乳酸杆菌制剂的保护机制。方法采用细菌培养法动态测定抗生素所致腹泻大鼠肠道菌群变化及肠系膜淋巴结、肝脏、脾脏和结肠组织的移位细菌量;应用光镜和电子显微镜观察肠黏膜组织超微结构变化。结果应用抗生素可致大鼠腹泻,肠道菌群失调,肠黏膜组织受损,发生肠道细菌移位。大肠埃希菌攻击可加重肠道菌群失调和肠黏膜损伤程度,促发细菌移位发生。乳酸杆菌可扶正肠菌群结构,修复损伤的肠黏膜,抑制肠细菌移位发生。结论阐明了抗生素、肠黏膜屏障功能、肠道菌群结构和肠道细菌移位间的互为因果,相互影响的关系。微生态制剂在维持机体微生态平衡、修复肠黏膜方面具有保护作用。     

鱼油对断奶大鼠脏器指数及肠道菌群影响的研究

目的探讨鱼油对断奶大鼠脏器指数及肠道菌群的影响。方法选取（21±3）d日龄断奶SD大鼠96只,雌雄各半,随机分为2组,实验组饲喂添加0．5％鱼油的饲料,对照组饲喂正常饲料。分别于第7、14、21、28天处死各组大鼠12只,分别测定大鼠肝脏指数、胸腺指数及脾脏指数;应用梯度稀释法和培养法测定大鼠4种肠道正常菌群,即肠杆菌、双歧杆菌、乳酸杆菌、葡萄球菌。结果实验组第7天脾脏指数与对照组相比差异有显著性（P〈0．05）,第28天肝脏指数和胸腺指数与对照组相比差异有显著性（P〈0．05）;实验组第7、14和21天肠道内肠杆菌数量与对照组相比差异有显著性（P〈0．05）,第7天大鼠肠道内葡萄球菌数量与对照组相比差异有显著性（P〈0．05）,第14天大鼠肠道内葡萄球菌数量与对照组相比差异有非常显著性（P〈0．01）,实验组大鼠肠道内乳酸杆菌和双歧杆菌数量与对照组相比虽有所增高,但差异无显著性（P〉0．05）。结论鱼油可以明显提高断奶大鼠的脏器指数,并且可通过增加断奶大鼠肠道内双歧杆菌和乳酸杆菌数量,降低肠道内肠杆菌和葡萄球菌数量,进而调节断奶大鼠的肠内菌群,改善肠道内环境。     

多联益生菌对5-氟尿嘧啶诱发大鼠肠道菌群紊乱及 肠黏膜损伤的保护作用研究

摘要：目的 研究多联益生菌对5-氟尿嘧啶（5-Fu）诱发大鼠肠道菌群紊乱、肠黏膜损伤以及脾脏损伤的保护作用。方法 28只SPF级SD大鼠随机分为正常组、5-Fu+生理盐水组、5-Fu+低剂量益生菌组和5-Fu+高剂量益生菌组,每组7只。所有大鼠于化疗结束后第3天处死,应用PCR-DGGE技术测定肠道菌群变化,鲎试剂测定血浆内毒素含量,HE染色观察肠黏膜变化和脾脏变化。结果 与5-Fu+生理盐水组相比,经益生菌干预后的大鼠其肠道菌群失调和体重减轻症状有明显改善,血浆内毒素含量显著降低,肠黏膜损伤也得到了显著改善,脾脏淋巴细胞明显增多,脾脏损伤程度降低。结论 该多联益生菌制剂能有效减轻5-Fu对大鼠肠黏膜及肠道菌群的破坏,同时减轻其对脾脏的损伤,并且改善大鼠在化疗后的生存质量。     

新生期大鼠补充罗伊乳杆菌DSM 17938 对肠道菌群及肠道发育的影响

目的探讨新生期应用罗伊乳杆菌DSM 17938对大鼠肠道菌群的影响。方法 24只自然分娩出生的新生SD大鼠随机分为对照组和益生菌组(每组12只),益生菌组大鼠在生后2 d(PND2)开始予罗伊乳杆菌DSM 17938[1×10~6 CFU/(g·bw),1次/d]灌胃,至出生后第6天(PND6),对照组同期予等量生理盐水灌胃。PND7及PND42时,两组分别处死6只SD大鼠,留取空肠、结肠黏膜及其肠内容物标本,采用16S rDNA V4区二代测序法检测空肠、结肠菌群构成;采用Image Pro Plus 6.0测量空肠、结肠绒毛长度及隐窝深度。结果益生菌组和对照组大鼠的体质量增量差异无统计学意义(t值分别为0.410、0.856,均P0.05),PND42时两组大鼠空肠、结肠黏膜绒毛长度及隐窝深度差异均无统计学意义(t值分别为1.796、0.115、1.122和0.715,均P0.05),两组大鼠空肠和结肠菌群的α多样性及β多样性差异在PND7及PND42时差异均无统计学意义(均P0.05),不同组间的肠道菌群LEfSe分析显示PND7及PND42时益生菌组和对照组大鼠肠道菌群组成无差异。结论自然分娩出生的健康SD大鼠生命早期添加罗伊乳杆菌DSM 17938不影响生命早期、远期肠道菌群的总体组成及远期肠道上皮组织结构的发育。     

不同品种、日龄和饲养方式对肉鸡肠道菌群的影响

目的研究品种、日龄和饲养方式对肉鸡肠道菌群的影响。方法选择北京油鸡和AA肉鸡各120只,分笼养和地面平养两种饲养方式,每种饲养方式60只鸡(5个重复,每个重复12只),于7、14、21和42日龄无菌收集十二指肠、回肠和盲肠内容物,分析各肠段肠道菌群变化。结果 (1)从十二指肠、回肠到盲肠微生物种类越来越丰富,相同肠段不同处理间随日龄的增加微生物种类也不断增加,十二指肠、回肠主要菌是植物乳酸杆菌和双歧杆菌,盲肠主要菌是拟杆菌和梭菌,且AA肉鸡拟杆菌数量大于北京油鸡;(2)品种对十二指肠和回肠条带数影响不显著,但北京油鸡大于AA肉鸡,日龄对回肠条带数的影响显著。品种与日龄的互作对盲肠的影响显著,品种、日龄和饲养方式三个处理的互作对回肠条带数的影响显著。(3)十二指肠各处理间肠道菌群的相似性在42日龄达到稳定,回肠在21日龄稳定,盲肠从7日龄到42日龄各处理间肠道菌群相似性变化不大。结论不同肠段的微生物区系有明显差异,品种、日龄和饲养方式对肉鸡肠道微生物区系有不同影响,但微生物区系会随着肉鸡日龄的增加趋于稳定。     

旋毛虫感染小鼠肠道菌群变化的研究

目的通过观察黑龙江株旋毛虫感染小鼠肠道分泌物中分泌型免疫球蛋白A、肠道菌群的变化,探讨感染小鼠肠道菌群的变化。方法分别于小鼠感染黑龙江株旋毛虫后7、14、21、28和35d,观察模型组及对照组小鼠肠道分泌物中的分泌型免疫球蛋白A、肠道双歧杆菌、乳酸杆菌、肠杆菌、肠球菌的菌群变化。sIgA采用放射免疫法检测。结果模型组sIgA分泌水平在感染后14d达高峰,随后缓慢下降但始终保持高水平（P〈0．01）。模型组肠道双歧杆菌的数量在感染后7d略低于对照组（P〈0．05）,第14天降至最低水平,随后逐渐升高,至感染后35d恢复正常水平。乳酸杆菌的数量在感染后7d略低于对照组,第14天降至最低水平,随后逐渐增加（P〈0．05）。肠杆菌的数量在感染后7d略高于对照组,感染后14d明显高于对照组,随后始终保持下降趋势（P〈0．05）。肠球菌在感染后7d略高于对照组（P〈0．05）,在14d明显高于对照组,随后缓慢下降,至感染后35d恢复正常水平。结论旋毛虫感染小鼠sIga的分泌在肠道免疫中发挥重要作用,同时也影响肠道菌群;肠道菌群的变化可能与旋毛虫感染小鼠免疫系统中sIgA的分泌有关。     

多联益生菌对5-氟尿嘧啶诱发大鼠肠道菌群紊乱及肠黏膜损伤的保护作用

目的研究多联益生菌对5-氟尿嘧啶(5-Fu)诱发大鼠肠道菌群紊乱、肠黏膜损伤以及脾脏损伤的保护作用。方法 28只SPF级SD大鼠随机分为正常组、5-Fu+生理盐水组、5-Fu+低剂量益生菌组和5-Fu+高剂量益生菌组,每组7只。所有大鼠于化疗结束后第3天处死,应用PCR-DGGE技术测定肠道菌群变化,鲎试剂测定血浆内毒素含量,HE染色观察肠黏膜变化和脾脏变化。结果与5-Fu+生理盐水组相比,经益生菌干预后的大鼠其肠道菌群失调和体重减轻症状有明显改善,血浆内毒素含量显著降低,肠黏膜损伤也得到了显著改善,脾脏淋巴细胞明显增多,脾脏损伤程度降低。结论该多联益生菌制剂能有效减轻5-Fu对大鼠肠黏膜及肠道菌群的破坏,同时减轻其对脾脏的损伤,并且改善大鼠在化疗后的生存质量。     

抗生素对新生大鼠肠道免疫发育影响及双歧杆菌干预的研究

目的 研究新生大鼠口服抗生素对肠道免疫发育的影响及双歧杆菌干预的效果.方法 选用50只7日龄新生SD大鼠,每组10只,随机分为5组:对照组(A)、抗生素组(B)、益生菌组(C)、益生菌干预组(D)和生理盐水组(E).A组为空白对照,B组给予头孢克洛灌胃,C组给予双歧杆菌灌胃,D组先给头孢克洛灌胃,2h后再灌长双歧杆菌,E组每天灌以等量的生理盐水,持续2周后处死大鼠,取少许新鲜盲肠内容物粪便涂片,免疫组化方法检测末端回肠肠组织中CD4、CD8的表达和组织学观察.结果 粪便涂片结果显示:B组与其余四组相比G+b占肠道总细菌数比率明显下降,G-b及G+c占总细菌数比率明显升高(P＜0.01).组织形态学观察:C组与E组肠黏膜绒毛和腺体发育良好,上皮结构完整,排列整齐,而A组腺体发育少,绒毛高度小;B组肠黏膜绒毛和腺体萎缩,黏膜水肿,部分上皮细胞变性、坏死、脱落;D组肠黏膜绒毛和腺体排列整齐绒毛显示清楚,少部分黏膜上皮细胞脱落、坏死.组肠组织中CD4、CD8的表达:B组CD4、CD8表达程度受到抑制,灰度值增大(P＜0.05);C组与E组相比CD4、CD8表达增加,灰度值比较差异有统计学意义(P＜0.05).结论 肠道菌群的正常定植,刺激肠道免疫系统的发育.新生大鼠口服抗生素后肠黏膜结构被破坏以及干扰肠道菌群的定植,影响肠道免疫系统的发育.长双歧杆菌的能预防抗生素引起的菌群紊乱,维持肠道黏膜的完整性,保护肠道免疫系统的正常发育.     

肝硬化患者肠道菌群失衡的研究现状

肝硬化是我国的一种常见病,近年来越来越多的研究表明肝硬化及其并发症(如门静脉高压、自发性腹膜炎、肝性脑病及肝癌等)都与肠道菌群失衡有着密切的联系。肝脏和肠道通过"肠-肝轴"紧密联系在一起,肝硬化时因小肠细菌过度生长、肠黏膜屏障功能受损、机体免疫功能下降等因素,导致细菌移位、肠道微生态失衡。而肠道微生态失衡又会使肝功能障碍进一步发展,引起肝性脑病等并发症。本文就目前国内外对肝硬化及其并发症与肠道细菌及真菌菌群失衡的研究进行综述。     

双歧三联活菌胶囊对结直肠癌术后肠道菌群及肠黏膜通透性的影响

目的探讨双歧三联活菌胶囊对结直肠癌术后肠道菌群及肠黏膜通透性的影响。方法选取拟行手术治疗的结直肠癌患者78例,随机分为观察组和对照组。两组患者术前常规禁食、肠道准备,采用开放根治性结/直肠癌切除手术治疗,术后予以围手术期常规治疗,并予以等氮量、等热量的营养支持。观察组术后第3天加用双歧三联活菌胶囊630mg/次,2次/d,连用7d。观察两组患者术前及治疗7d后肠道菌群(双歧杆菌、乳酸杆菌、大肠埃希菌和粪肠球菌)及肠黏膜通透性指标[血清二胺氧化酶(DAO)]的变化,并比较感染并发症的发生率。结果治疗7d后,两组患者双歧杆菌、乳酸杆菌和粪肠球菌数量较前明显下降,大肠埃希菌数量较前明显上升(t=2.17、2.25、2.21、2.32、2.89、3.08、2.97、3.12,P0.05或P0.01),且观察组下降或上升的幅度明显小于对照组(t=2.14、2.25、2.18、2.23,P0.05);两组血清DAO水平均较前明显上升(t=2.27、3.21,P0.05或P0.01),且观察组上升幅度明显小于对照组(t=2.23,P0.05);同时观察组患者感染并发症的总发生率明显低于对照组(χ2=5.03,P0.05)。结论结直肠癌术后存在一定程度的肠道菌群的紊乱,肠黏膜通透性上升和肠道屏障功能受损。双歧三联活菌胶囊用于结直肠癌术后可纠正与调节肠道菌群的紊乱,降低肠黏膜通透性,保护与修复肠黏膜屏障功能,从而减少或避免肠道细菌和内毒素的移位,降低感染并发症的发生。     

An early stimulation of solid feed intake slightly influences the morphological gut maturation in the rabbit

The impact of dietary factors on the gut morphological maturation is poorly documented in rabbits. The weights of the digestive segments as well as the morphology of villi and crypts along the small intestine were analysed weekly from day 14 till day 49, in two rabbit groups weaned at either 21 (W21 group, n = 12 litters) or 35 days (W35 group, n = 12 litters) of age. From 21 till 35 days, the W21 group ate 57% more solid feed than the W35 group (P < 0.01), and presented slighter body weights from day 28 till day 49 (-9%, P < 0.05). Tissue weights of the empty digestive segments, as expressed relative to the body weights, were higher in the W21 than in the W35 group from day 28 till day 49 (P < 0.001), whereas absolute tissue weights appeared similar (except for the proximal colon). From day 28 to day 49, small intestinal villi grew in height and surface area (P < 0.05) whereas the crypts deepened. Villous height followed a proximo-distal decreasing gradient from the duodenum to the ileum (P < 0.05) from day 28 onward. The villous height to width ratio changed with the beginning of significant solid feed intake: from a thin shape until day 21, villi became wider from day 28 on. The effect of weaning age on mucosal morphology was insignificant, except for the jejunal crypts whose surface area and depth were higher in the W21 group. The present results showed that morphological changes in the digestive tract of young rabbits were weakly influenced by an early stimulation of solid feed intake.     

Effects of Enterococcus faecium and Bacillus cereus var. toyoi on the morphology of the intestinal mucous membrane in piglets

Eighty piglets aged 14, 28, 35 and 56 days — weaned at day 28 — were subjected to this investigation. Each age-group consisted of five animals which were fed an Enterococcus faecium NCIMB 10415 (Cylactin®) and Bacillus cereus var. toyoi (Toyocerin®) based diet. Five animals served as controls. Tissue samples were collected immediately after sacrifice at 8.30 h a.m. from duodenum, jejunum, ileum, cecum and colon to examine intestinal morphology and histochemistry. The results showed that with respect to villus height and crypt depth supplementation of probiotics in piglets feed seemed to influence the morphology and enlargement factor not at all or only to a certain extent. With respect to the number of goblet cells, the difference between probiotic fed animals and control animals was generally extremely low. The shape of the villi of the small intestinal segments greatly varied in all age groups of control and probiotic fed animals. However, this morphological variety does not depend on the mode of feeding.     

Changes in the small intestine of Schistosoma mansoni-infected mice fed a high-fat diet

The consumption of a high-fat diet modifies both the morphology of the small intestine and experimentally tested effects of schistosomiasis mansoni. However, whether a schistosomiasis infection associated with a high-fat diet causes injury to the small intestine has never been investigated. Mice were fed either a high-fat or a standard-fat diet for 6 months and were then infected with Schistosoma mansoni cercariae. Physical characteristics of the intestinal tissue (mucosal thickness, small intestinal villi length and height, and abundance of goblet cells and enterocytes on the villous surface) and the distribution of granulomas along the intestinal segments and their developmental stage were measured at the time of sacrifice (9 or 17 weeks post-infection). The group fed a high-fat diet exhibited different granuloma stages, whereas the control group possessed only exudative granulomas. The chronically infected mice fed a high-fat diet exhibited higher granuloma and egg numbers than the acutely infected group. Exudative, exudative/exudative-productive and exudative-productive granulomas were present irrespective of diet. Computer-aided morphometric analysis confirmed that villus length, villus width, muscular height and submucosal height of the duodenal and jejunal segments were affected by diet and infection. In conclusion, a high-fat diet and infection had a significant impact on the small intestine morphology and morphometry among the animals tested.     

Toxic Effects of Maternal Zearalenone Exposure on Intestinal Oxidative Stress,Barrier Function,Immunological and Morphological Changes in Rats

The present study was conducted to investigate the effects of maternal zearalenone (ZEN) exposure on the intestine of pregnant Sprague-Dawley (SD) rats and its offspring. Ninety-six pregnant SD rats were randomly divided into four groups and were fed with diets containing ZEN at concentrations of 0.3 mg/kg, 48.5 mg/kg, 97.6 mg/kg or 146.0 mg/kg from gestation days (GD) 1 to 7. All rats were fed with mycotoxin-free diet until their offspring were weaned at three weeks of age. The small intestinal fragments from pregnant rats at GD8, weaned dams and pups were collected and studied for toxic effects of ZEN on antioxidant status, immune response, expression of junction proteins, and morphology. The results showed that ZEN induced oxidative stress, affected the villous structure and reduced the expression of junction proteins claudin-4, occludin and connexin43 (Cx43) in a dose-dependent manner in pregnant rats. Different effects on the expression of cytokines were also observed both in mRNA and protein levels in these pregnant groups. Ingestion of high levels of ZEN caused irreversible damage in weaned dams, such as oxidative stress, decreased villi hight and low expression of junction proteins and cytokines. Decreased expression of jejunal interleukin-8 (IL-8) and increased expression of gastrointestinal glutathione peroxidase (GPx2) mRNA were detected in weaned offspring, indicating long-term damage caused by maternal ZEN. We also found that the Nrf2 expression both in mRNA and protein levels were up-regulated in the ZEN-treated groups of pregnant dams and the high-dose of ZEN group of weaned dams. The data indicate that modulation of Nrf2-mediated pathway is one of mechanism via which ZEN affects gut wall antioxidant and inflammatory responses.     

Age-dependent development in protein digestibility and intestinal morphology in weaned pigs fed different protein sources

Today, weaner diets are optimised using digestibility coefficients obtained from grower-finisher pigs, which may overestimate the digestibility in weaners. The aim of this study was to evaluate the standardised ileal digestibility (SID) of CP and amino acids (AAs), and the intestinal morphology in pigs 0–4 weeks postweaning when fed different protein sources. The experiment included 128 pigs weaned at day 28 and the protein sources were wheat, soybean meal (SBM), enzyme-treated soybean meal (ESBM), hydrothermally treated rapeseed meal (HRSM) and casein. The experiment was conducted as a difference method study including wheat in all diets. Eight pigs were slaughtered on the day of weaning (day 0) and six pigs/treatment were slaughtered at days 7, 14, 21, and 28 postweaning. The SID of CP and AA, as average over the four weeks, was lowest for ESBM and highest for wheat and casein, with SBM and HRSM being intermediate. The SID of CP and AA increased (both linear and quadratic, P < 0.05) over time after weaning. The average SID of CP for all protein sources postweaning was 0.38, 0.59, 0.76, and 0.71 on days 7, 14, 21, and 28, respectively. These differences were significant (P < 0.05) between days 7 and 21, and between days 7 and 28 (P < 0.05), whereas there tended to be a difference between days 7 and 14 (P = 0.06). Protein source did not affect the small intestinal morphology response parameters, whereas time after weaning did. Villous height and villous height to crypt depth ratio differed (P < 0.05) between the days 0 and 7, with shorter villi and a higher ratio at day 7. Crypt depth was not altered between days 0 and 7, or between days 7 and 14. For villi density, crypt density and small intestinal length, a significant increase from days 7 to 14 was observed, but there was no further increase to or difference between days 21 and 28. In conclusion, the low SID of CP in casein on day 7 (0.50) illustrates the challenges related to protein digestion in weanling pigs. The SID of CP and AA is very low during the first two weeks postweaning and time after weaning is more important for protein digestibility, than the source of protein. Fewer mature epithelial cells and less absorptive area in the small intestine in the early postweaning period may partly explain the poor protein digestibility.     

Observation of mucosal pathology after praziquantel treatment in experimental Fibricola seoulensis infection in rats

It is well known that the duodenum of mice or rats infected with Fibricola seoulensis shows atrophy of villi (shortening, blunting, widening, fusion) and hyperplasia of crypts. This study was performed to observe healing process of these pathological changes after deworming with anthelmintic treatment. Albino rats infected each with 1,000 metacercariae of F. seoulensis were treated with single dose of 10 mg/kg praziquantel on day 15 post-infection. On day 1, 3, 5, 7, 15, 21 and 28 after the treatment, they were sacrificed and their duodenums were histopathologically studied. Control (uninfected) rats showed their normal finger-like projections of duodenal villi and well arranged crypts. In comparison, untreated (infected) controls revealed severe mucosal changes characteristic of villous atrophy and crypt hyperplasia in their duodenum. The damaged duodenal mucosa was found to restore its normal morphology after praziquantel treatment; until day 3 post-treatment the mucosa was severely atrophied; on day 5 long and slender villi sometimes appeared among the fused and stout ones; after day 15 the villi were in their normalizing process. From this experiment, it was shown that the mucosal changes in the duodenum of rats caused by F. seoulensis infection were completely reversible in 21-28 days after anthelmintic treatment.     

Insights into Embryo Defenses of the Invasive Apple Snail Pomacea canaliculata: Egg Mass Ingestion Affects Rat Intestine Morphology and Growth

Background

The spread of the invasive snail Pomacea canaliculata is expanding the rat lungworm disease beyond its native range. Their toxic eggs have virtually no predators and unusual defenses including a neurotoxic lectin and a proteinase inhibitor, presumably advertised by a warning coloration. We explored the effect of egg perivitellin fluid (PVF) ingestion on the rat small intestine morphology and physiology.

Methodology/Principal Findings

Through a combination of biochemical, histochemical, histopathological, scanning electron microscopy, cell culture and feeding experiments, we analyzed intestinal morphology, growth rate, hemaglutinating activity, cytotoxicity and cell proliferation after oral administration of PVF to rats. PVF adversely affects small intestine metabolism and morphology and consequently the standard growth rate, presumably by lectin-like proteins, as suggested by PVF hemaglutinating activity and its cytotoxic effect on Caco-2 cell culture. Short-term effects of ingested PVF were studied in growing rats. PVF-supplemented diet induced the appearance of shorter and wider villi as well as fused villi. This was associated with changes in glycoconjugate expression, increased cell proliferation at crypt base, and hypertrophic mucosal growth. This resulted in a decreased absorptive surface after 3 days of treatment and a diminished rat growth rate that reverted to normal after the fourth day of treatment. Longer exposure to PVF induced a time-dependent lengthening of the small intestine while switching to a control diet restored intestine length and morphology after 4 days.

Conclusions/Significance

Ingestion of PVF rapidly limits the ability of potential predators to absorb nutrients by inducing large, reversible changes in intestinal morphology and growth rate. The occurrence of toxins that affect intestinal morphology and absorption is a strategy against predation not recognized among animals before. Remarkably, this defense is rather similar to the toxic effect of plant antipredator strategies. This defense mechanism may explain the near absence of predators of apple snail eggs.