Objective:To investigate the intracellular localization and binding of duck invariant chain (Ii) to major histocompatibility complex (MHC) class Iα or MHC class IIβ molecules. Methods:First, the genes of duck Ii, MHC Iα, and MHC IIβ were amplified by PCR, cloned into expression plasmids, and then sequenced to compare homology between ducks and selected agricultural species. Second, Rosetta or 293T cells were transfected with plasmids encoding Ii, MHC Iα, and MHC IIβ either alone or in combination to interrogate binding following expression. The fluorescent reporter molecules used to tag/image transfected cells and expressed protein were identified by western blot. Additionally, the binding interaction of the expressed products was observed by affinity purification using pull-down assay and coimmunoprecipitation. Finally, intracellular colocalization was observed by laser confocal microscopy. Results:The results of western blots demonstrated that all constructed plasmids successfully expressed proteins. Pull down and coimmunoprecipitation revealed that Ii could bind Iα or IIβ, forming Ii-Iα or Ii-IIβ complexes, and they dissociated into single molecules after SDS treatment. In 293T cells, Ii interacts with MHC Iα or MHC IIβ chains and colocalizes intracellularly. Confocal fluorescence microscopy assay indicated that MHC Iα and IIβ were located in the endoplasmic reticulum and endosome. Conclusions:Both MHC and Ii proteins exhibit direct interactions and can colocalize in the endoplasmic reticulum and endosomes. Clinical Relevance:This study investigates the relationship between duck Ii and MHC proteins, offering a theoretical foundation for exploring the role of Ii in immune responses and advancing the development of Ii-vectored avian vaccines.
Duck adenovirus type-3 (DAdV-3) infections have severe effects on duck health, and rapid detection methods are crucial to reduce the morbidity and mortality associated with this pathogen in clinical practice. In this study, we established, optimized, and validated a novel recombinase polymerase amplification (RPA)-lateral flow dipstick (LFD) assay for the detection of DAdV-3. Next, we established a clinical infection model based on the pathogenicity of the DAdV-3 strain and tested the effectiveness of the RPA-LFD assay in controlling an outbreak of DAdV-3 infection. The findings indicated that the RPA-LFD assay could be performed within 30 min at 42°C. Specificity tests indicated no cross-reactivity with other viruses. The detection limit of the assay was 1 × 101 copies/μL. We evaluated 65 clinical samples using RPA-LFD and quantitative polymerase chain reaction (qPCR), and both methods showed a positivity rate of 33.8% and a coincidence rate of 100%. The kappa (κ) value of the RPA-LFD and qPCR assays was 1 (p < 0.001). The application of this assay in experimentally infected ducklings reduced the mortality rate from 20 to 8%. Thus, the RPA-LFD assay established in this study demonstrated high specificity, sensitivity, rapidity, and efficacy, indicating its potential for rapid detection of DAdV-3 in clinical settings.
Duck adenovirus Type 3 (DAdV-3) severely affects the health of ducks; however, its pathogenicity in chickens remains unknown. The objectives of this study were to evaluate the pathogenicity and major pathological changes caused by DAdV-3 in chickens. Viral DNA was extracted from the liver of the Muscovy duck, and the fiber-2 and hexon fragments of DAdV-3 were amplified through polymerase chain reaction (PCR). The evolutionary tree revealed that the isolated virus belonged to DAdV-3, and it was named HE-AN-2022. The mortality rate of chicks that received inoculation with DAdV-3 subcutaneously via the neck was 100%, while the mortality rate for eye–nose drop inoculation was correlated with the numbers of infection, with 26.7% of chicks dying as a result of exposure to multiple infections. The main symptoms exhibited prior to death were hepatitis–hydropericardium syndrome (HHS), ulceration of the glandular stomach, and a swollen bursa with petechial hemorrhages. A histopathological examination revealed swelling, necrosis, lymphocyte infiltration, and basophilic inclusion bodies in multiple organs. Meanwhile, the results of quantitative real-time PCR (qPCR) demonstrated that DAdV-3 could affect most of the organs in chickens, with the gizzard, glandular stomach, bursa, spleen, and liver being the most susceptible to infection. The surviving chicks had extremely high antibody levels. After the chickens were infected with DAdV-3 derived from Muscovy ducks, no amino acid mutation was observed in the major mutation regions of the virus, which were ORF19B, ORF66, and ORF67. On the basis of our findings, we concluded that DAdV-3 infection is possible in chickens, and that it causes classic HHS with ulceration of the glandular stomach and a swollen bursa with petechial hemorrhages, leading to high mortality in chickens. The major variation domains did not change in Muscovy ducks or in chickens after infection. This is the first study to report DAdV-3 in chickens, providing a new basis for preventing and controlling this virus.
恒定链(invariant chain,Ii)是重要的免疫分子.研究发现Ii不仅是MHCII类分子的伴侣分子,而且具有多种功能.本文首先概述了 Ii的基本结构、主要功能、胞内组装和转运、Ii作为载体的应用.其次,归纳和总结了近几年关于Ii的最新研究进展:Ii与MHC分子互相作用的功能结构域及其在细胞的定位特征、调节T和B等免疫细胞的功能与途径、Ii与巨噬细胞迁移因子相互作用从而启动炎症反应的信号通路和相应结构域;与Ii关联的多种酶类和生物活性因子及其作用特点、与Ii相关的疾病;基于Ii结构的疫苗载体及其增强免疫的机理;以及我国在畜禽、鱼类上的研究成果.最后,展望了 Ii在理论研究和畜牧兽医领域实际应用的发展趋势.本文从宏观和微观方面进行阐述,了解、认识Ii在动物免疫中的多功能作用及其机理的科学知识和最新进展,为推动相关研究提供有益借鉴和参考.
Abstract Background Duck adenovirus type-3 (DAdV-3) seriously affects the health of ducks, however, the pathogenicity of the virus in chickens remains unknown. The objectives of the study were to evaluate the pathogenicity and major pathological changes caused by DAdV-3 in chickens. Results The specific fragment of the virus was amplified by polymerase chain reaction (PCR), and the evolutionary tree showed that the isolated virus belonged to DAdV-3, named HE-AN-2022. The mortality rate of chicks was 100% after subcutaneous inoculation at the neck, while the mortality rate of eye-nose drop inoculation was correlated with the number of infections, with 26.7% of chicks dying if exposed to multiple infections. The main symptoms of chicks before death were hepatitis-hydropericardium syndrome (HHS), ulceration of the glandular stomach and swollen bursa with petechial hemorrhages. Histopathological examination showed swelling, necrosis, lymphocyte infiltration and alkaline inclusion bodies in multiple organs. The results of quantitative real-time PCR (qPCR) showed that DAdV-3 could infect most organs of chickens, and the gizzard, glandular stomach, bursa, spleen and liver carried the highest amount of virus. Surviving chicks produced extremely high antibody levels. After infecting chickens with DAdV-3 derived from Muscovy ducks, no amino acid mutation was observed in the major mutation regions ORF19B, ORF66 and ORF67 of the virus. Conclusion DAdV-3 can infect chickens, causing classic HHS with ulceration of the glandular stomach and swollenbursa with petechial hemorrhages, leading to high mortality in chickens.The major variation domains did not change after infection with the virus in Muscovy ducks and chickens. This is the first study to report the of DAdV-3 in chickens, and this study provides a new basis for the prevention and control of the virus.
通过建立雏鹅高尿酸血症模型,研究高蛋白日粮对雏鹅血清尿酸水平、肝脏和肾脏超微结构及AB-CG2表达的影响.超微结构观察发现,对照组(CP)肝脏和肾脏细胞器结构完好,高蛋白1组(HP1)、高蛋白2组(HP2)肝脏部分线粒体开始出现肿胀、空泡变性,内质网肿胀消失,糖原减少.肾脏细胞部分线粒体开始出现肿胀、空泡变性,核膜间隙增大.雏鹅7日龄时,HP2组鹅的血清尿酸水平极显著高于CP组和HP1组(P<0.01);雏鹅14日龄时,HP2组鹅的血清尿酸水平极显著高于CP组和HP1组(P<0.01),HP1组鹅血清尿酸水平极显著高于CP组(P<0.01).qPCR、Western-boltting及免疫组化结果显示,HP2组鹅十二指肠、空肠中ABCG2基因和蛋白的表达显著或极显著高于CP组和HP1组(P<0.05或P<0.01),鹅肾脏ABCG2基因和蛋白的表达显著或极显著低于CP组和HP1组(P<0.05或P<0.01);HP1组鹅ABCG2基因和蛋白在空肠中的表达显著高于CP组(P<0.05),在肾脏中的表达显著低于CP组(P<0.05).综上所述,饲喂高蛋白日粮导致雏鹅肝脏、肾脏细胞受损,血清尿酸水平升高,引发雏鹅高尿酸血症,空肠、十二指肠中ABCG2的表达增加,而肾脏中ABCG2的表达降低.
[目的]探究益生菌对肉仔鸡血清免疫球蛋白与肠道Toll样受体通路的影响.[方法]选择90只1日龄白羽肉杂鸡,随机分为3组,即对照组、益生菌低剂量组和益生菌高剂量组.其中对照组饲喂基础日粮,益生菌低剂量组在每千克基础日粮中添加复合益生菌109 cfu,益生菌高剂量组在每千克基础日粮中添加复合益生菌2×109 cfu.第21天,每组随机选取10只鸡放血致死,并立即采集样本,进行ELISA、免疫组化、qPCR及Western blot检测.[结果]在第21天,益生菌低剂量组和益生菌高剂量组鸡的料肉比分别比对照组降低9%和12%(P<0.05),血清IgG含量分别升高28%和40% (P<0.01),血清IgM含量分别升高44%和58% (P<0.01).益生菌低剂量组TLR4蛋白表达和mRNA相对表达量比对照组分别升高33%和28%,AP-1蛋白表达和mRNA相对表达量分别比对照组升高106%和67% (P<0.01);益生菌高剂量组TLR4蛋白表达及mRNA相对表达量比对照组分别升高106%和69%,AP-1蛋白表达及mRNA相对表达量比对照组分别升高163%和98% (P<0.01).[结论]饲喂益生菌可降低肉仔鸡料肉比,提高血清中IgG、IgM的含量,并且通过调节Toll样受体通路蛋白表达可以提高机体免疫力.
[目的]探讨高蛋白日粮对雏鹅生长性能、血清炎症因子及肝脏、肾脏、空肠、十二指肠中黄嘌呤脱氢酶(XDH)表达的影响.[方法]选用1日龄雁鹅72只,随机分成对照组(CP)、高蛋白组1(HP1)和高蛋白组2(HP2),分别饲喂含180.00,230.00和280.00 g/kg粗蛋白的日粮,每组3个重复,每个重复8只鹅.试验为期14 d,每周测定各组雏鹅采食量、体质量.于7和14 d采血,分离血清,用ELISA法测定血清中黄嘌呤氧化酶(XOD)活性及肿瘤坏死因子(TNF-α)、白细胞介素1β(1L-1β)、白细胞介素6(IL-6)、尿酸(UA)含量;采用RT-qPCR、免疫组化法、West-ern-blotting法测定14日龄雏鹅肝脏、肾脏、空肠、十二指肠中XDH基因及其蛋白的表达水平.[结果])7日龄时,HP2组雏鹅的平均体质量显著小于CP组(P<0.05),极显著小于HP1组(P<0.01),HP1组雏鹅的平均日增重显著大于HP2组(P<0.05);14日龄时,CP组雏鹅的平均体质量、平均日增重、日采食量极显著高于HP1和HP2组(P<0.01).2)7日龄时,HP2组雏鹅血清中的1L-1β、TNF-α、UA含量及XOD活性均极显著高于CP组(P<0.01),1L-1β、TNF-α、UA含量显著高于HP1组(P<0.05);HP1组雏鹅血清中TNF-α质量浓度极显著高于CP组(P<0.01).14日龄时,HP2组雏鹅血清中1L-1β、IL-6、TNF-α、 UA含量及XOD活性均显著或极显著高于CP组和HP1组;HP1组雏鹅血清中的1L-1β、IL-6、TNF-α、UA含量及XOD活性极显著高于CP组(P<0.01).3)HP2组雏鹅肝脏、肾脏、十二指肠的XDH基因及其蛋白的表达显著或极显著高于CP组(P<0.05或P<0.01),肝脏、肾脏中XDH基因及其蛋白的表达显著或极显著高于HP1组(P<0.05或P<0.01);HP1组雏鹅肝脏、十二指肠中XDH基因及其蛋白的表达显著或极显著高于CP组(P<0.05或P<0.01).各组雏鹅空肠中XDH基因及其蛋白的表达无显著差异(P>0.05).[结论]摄入过高水平的蛋白会导致雏鹅生长性能降低,机体发生炎症反应,肝脏、肾脏、十二指肠中XDH基因和蛋白表达量增加,从而引发雏鹅高尿酸血症.
[目的]采用色谱-质谱联用的代谢组学方法分析高蛋白日粮对雏鹅血清中内源性小分子代谢产物的影响.[方法]选用1日龄雁鹅48只,随机分成2组,每组24只鹅.对照组(CP)饲喂基础日粮(含18%粗蛋白),高蛋白组(HP)饲喂含23%粗蛋白的日粮,试验期为14 d.[结果]:与对照组相比,雏鹅7、14日龄时,高蛋白组鹅血清中谷丙氨酸氨基转移酶、天门冬氨酸氨基转移酶、乳酸脱氢酶活性和总蛋白、尿酸含量极显著升高(P<0.01),球蛋白含量显著升高(P<0.05);14日龄时,高蛋白组鹅血清中总胆红素、间接胆红素、尿素、肌酐含量极显著升高(P<0.01).正交偏最小二乘-判别分析法((OPLS-DA))分析结果显示,2组间有明显的分离趋势,提示HP组与CP组血清代谢谱有差异.对LC-TOF/MS和GC-TOF/MS分析结果进行代谢富集和途径分析表明,氨酰tRNA生物合成、嘌呤代谢、嘧啶代谢、β-丙氨酸代谢、赖氨酸降解、磷酸肌醇代谢、泛酸和辅酶A生物合成途径发生了显著改变.热图分析显示高蛋白组和对照组差异代谢物含量显著不同.[结论]本试验通过非靶向代谢组学筛选出的雏鹅血清中黄嘌呤、瓜氨酸、尿酸等41种代谢物与高蛋白日粮诱导雏鹅高尿酸血症发生相关,为潜在标志物.
本研究旨在探明鸡恒定链(invariant chain,Ii)与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征.首先,用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii(Cyt-Tra)]、Ii CLIP(classⅡ-associated invar-iant chain peptide)-三聚体区[Ii(CLIP-TRIM)]和Ii突变体[Ii(M81-87aa)、Ii(M91-99aa)和Ii(M81-99aa)]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒.其次,将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T,观察它们的共定位.将构建的原核重组质粒进行表达和纯化,最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域.结果 表明,成功构建Ii结构域及Ii突变体的重组质粒.Ii(Cyt-Tra)及Ii突变体均能与Rab5a和Rab7b在细胞内共定位,而Ii(CLIP-TRIM)与空载体却不能.Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域,而不是CLIP与三聚体区.综上所述,鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区,而不是内质网腔区.这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制,为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径.
The aim of the present study was to investigate the immunoregulatory effects of Astragalus polysaccharide (APS) on RAW264.7 cells. The production of cytokines by RAW264.7 cells was analyzed using ELISA, while cell viability and optimal concentration of APS were assessed using the Cell Counting Kit-8 assay. In addition, the mRNA levels of IL-6, inducible nitric oxide synthase (iNOS) and TNF-alpha were determined by reverse transcription-quantitative PCR analysis. The levels of co-stimulatory molecules and cell cycle distribution were assessed by flow cytometry. Electrophoretic mobility shift assay was used to determine the effects of APS on p65 expression. Compared with controls, APS enhanced the production of NO, the gene expression of TNF-alpha, IL-6 and iNOS and the protein levels of phosphorylated p65, p38, Jun N-terminal kinase and extracellular signal regulated kinase in RAW264.7 cells, whereas these effects of APS were alleviated by pyrrolidine dithiocarbamate. The results of the present study indicated that the immunoregulatory effects of APS are mediated, at least in part, via the activation of the NF-kappa B p65/MAPK signaling pathway.
[目的]研究高蛋白饲粮对肉鸡肾氧化损伤及黄嘌呤氧化酶(xanthine oxidase,XOD)的影响.[方法]选取120羽20日龄健康白羽肉杂鸡,随机分为4组,每组5个重复,每个重复6羽.对照组饲喂基础饲粮(蛋白质含190 g/kg),试验组饲喂在基础饲粮中添加豆粕使粗蛋白含量分别达到220,250,280 g/kg的饲粮.试验期为14 d.于试验第0,7和14天,分别从每组随机选取5羽鸡经翅下静脉采血,分离血清,用于肾功能(血清钙(Ca)、肌酐(CREA)、尿素氮(BUN)、尿酸(UA)、血清磷(P)、镁(Mg)含量)、脂质过氧化(血清丙二醛(MDA)、一氧化氮(NO)、羟自由基(? OH)含量和总抗氧力(T-AOC)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活性)及XOD活性检测.试验结束时,每组选5羽鸡进行解剖,取肾脏组织,用于检测XOD基因mRNA的表达量.[结果]1)与对照组相比,各高蛋白饲粮组肉鸡血清Ca、CREA、BUN、UA含量显著或极显著上升(P<0.05或P<0.01),血清P、Mg含量变化不显著(P>0.05).2)与对照组相比,各高蛋白饲粮组肉鸡血清MDA、NO、?OH含量显著或极显著升高(P<0.05或P<0.01),T-AOC、GSH-Px、SOD活性显著或极显著降低(P<0.05或P<0.01).3)与对照组相比,各高蛋白饲粮组XOD活性显著升高(P<0.05),肾组织XOD mRNA表达量显著增加(P<0.05).[结论]高蛋白饲粮可改变肉鸡肾功能与脂质过氧化,导致血清XOD活性以及肾组织XOD mRNA表达量升高,导致鸡肾脏损伤.
随机选取体重相近[(12.14±0.59) kg]的3月龄白山羊断奶羔羊21只,断奶时间不超过3d,随机分为对照组、低浓度(0.625 g· kg-1)茶皂素组和高浓度(2.5 g· kg-1)茶皂素组,每组7只,饲喂56 d后采集小肠上皮组织,检测其形态结构变化,并取小肠内容物,分析其微生物组成,探究茶皂素对安徽白山羊断奶羔羊小肠黏膜形态和肠道菌群的影响.结果 表明:①与对照组相比,添加茶皂素极显著增加十二指肠、空肠、回肠绒毛长度及绒毛长度/隐窝深度的比值,极显著降低隐窝深度.②与对照组相比,茶皂素组广古菌门、拟杆菌门、疣微菌门、红椿杆菌纲、产芽孢菌纲、双歧杆菌目、甲烷杆菌目、双歧杆菌科和甲烷杆菌科相对丰度显著增加,茶皂素组放线菌纲相对丰度显著下降;与低浓度茶皂素组相比,高浓度茶皂素组红蝽菌目、红蝽菌科和丹毒丝菌科相对丰度显著升高.综上所述,茶皂素能促进白山羊羔羊小肠各段发育,提高肠道有益菌群丰度,降低有害菌群比例,改善肠道菌群结构.
ABSTRACT β-Conglycinin and glycinin are known to induce various allergic reactions, however, but little is known about the mechanism underlying the development of allergy to soybean antigen proteins. In this study, porcine intestinal epithelial cells (IPEC-J2) were used to investigate the effects of soybean antigen proteins, β-conglycinin and glycinin, on cells to determine whether the caspase-3/mitochondrion-regulated apoptotic pathway underlies the allergic reaction. IPEC-J2 cells were treated with different concentrations (0, 5, and 10 mg mL−1) of β-conglycinin or glycinin, and 50 μM z-DEVD-FMK (a caspase-3 inhibitor). The results show that the apoptosis rate, mitochondrion-regulated mRNA and protein expression levels, caspase-3 activation, cyt-c release, cytoskeleton and tight junction protein expression, mitochondrial membrane potential depolarization and mitochondrial injury were significantly aggravated with cultured in increasing concentrations of β-conglycinin or glycinin. While these effects were inhibited by application of the caspase-3 inhibitor. Thus, we concluded that β-conglycinin and glycinin cause IPEC-J2 cell apoptosis via the caspase-3/mitochondrion-regulated apoptotic pathway.
为建立快速检测鹅星状病毒(GAstV)的方法,本研究根据GenBank中GAstV ORF2基因序列设计1对特异性引物,构建重组质粒pMD19-T-GAstV,以其作为标准品建立了GAstV的SYBR Green I荧光定量RT-PCR检测方法.优化其反应条件及体系,进行特异性、敏感性和重复性试验以及临床样本检测.试验结果显示,除GAstV外,禽白血病病毒(ALV)、血清4型禽腺病毒(FAdV-4)、鸡传染性喉气管炎病毒(ILTV)和鸡传染性贫血病病毒(CIAV)等常见禽病病原利用该方法检测均呈阴性,表明其特异性良好;建立的荧光定量RT-PCR检出的最低模板含量为1μl 3.75×101拷贝,敏感性较高;组内和组间重复性试验变异系数均小于1%.利用该方法对来自安徽地区的32份临床样品进行检测,阳性检出率为43.75%,常规PCR方法阳性检出率为18.75%,阳性检出符合率100%,表明该方法可用于临床样品检测.该方法的建立为临床样品中GAstV的快速高效检测提供了技术支持.
[目的]旨在研究饲喂高钙、高蛋白日粮对肉仔鸡生长性能、血清生化指标、抗氧化指标及盲肠菌群的影响.[方法]选取160只体质量相近的健康1日龄肉仔鸡,随机分为4组,每组5个重复,每个重复8只,分别为对照组(CP含量21%,Ca含量1%)、高蛋白组(CP含量31%,Ca含量1%)、高钙组(CP含量21%,Ca含量5%)和高钙高蛋白组(CP含量31%,Ca含量5%).[结果]结果表明:(1)仔鸡21日龄时,对照组鸡的平均体质量、日增质量、日采食量均极显著高于其他3组(P<0.01),料重比极显著小于高钙高蛋白组(P<0.01);高蛋白组鸡的平均体质量、日采食量显著高于高钙组和高钙高蛋白组(P<0.05);高钙组鸡的平均体质量、日增质量显著高于高钙高蛋白组(P<0.05)而料重比显著小于高钙高蛋白组(P<0.05).(2)高钙、高蛋白日粮可导致仔鸡血清中AST、ALT、UA、CREA-S、UREA、GSH-Px、SOD、MDA、T-AOC、XOD活性或含量异常.(3)IlluminaMiseq高通量测序结果表明,仔鸡盲肠中的厚壁菌门Firmicutes、变形菌门Proteobacteria、厌氧杆菌属Barnesiella、另枝菌属Alistipes、粪便杆菌属Faecalibacterium、理研菌属Rikenella、卟啉单胞菌属Porphyromonas、梭状芽孢杆菌属第十八类群Clostridium_ⅩⅧ、嗜碱菌Alkaliphilus、颤杆菌克属Oscillibacter、梭状芽孢杆菌Clostridium的物种丰富度发生显著变化(P<0.01或P<0.05);PICRUSt功能预测结果显示,葡萄糖和一些氨基酸和核苷酸的合成代谢出现紊乱.(4)对照组鸡肝肾组织正常,结构清晰;其他三组鸡肝肾组织发生不同程度的病变.[结论]高钙高蛋白日粮导致仔鸡生长性能、抗氧化能力降低,肝肾功能受损,引起仔鸡高尿酸血症及盲肠菌群结构异常.
Green tea polyphenols (GTPs) exhibit beneficial effects towards obesity and intestinal inflammation; however, the mechanisms and association with gut microbiota are unclear. We examined the role of the gut microbiota of GTPs treatment for obesity and inflammation. Canines were fed either a normal diet or high-fat diet with low (0.48% g/kg), medium (0.96% g/kg), or high (1.92% g/kg), doses of GTPs for 18 weeks. GTPs decreased the relative abundance of Bacteroidetes and Fusobacteria and increased the relative abundance of Firmicutes as revealed by 16S rRNA gene sequencing analysis. The relative proportion of Acidaminococcus, Anaerobiospirillum, Anaerovibrio, Bacteroides, Blautia, Catenibactetium, Citrobacter, Clostridium, Collinsella, and Escherichia were significantly associated with GTPs-induced weight loss. GTPs significantly (P<.01) decreased expression levels of inflammatory cytokines, including TNF-α, IL-6, and IL-1β, and inhibited induction of the TLR4 signaling pathway compared with high-fat diet. We show that the therapeutic effects of GTPs correspond with changes in gut microbiota and intestinal inflammation, which may be related to the anti-inflammatory and anti-obesity mechanisms of GTPs.
[目的]探讨高钙高蛋白日粮对海兰褐蛋鸡肝脏的影响.[方法]将80只40日龄海兰褐蛋鸡随机均分为4组,1组饲喂基础日粮(对照组),2、3和4组分别饲喂高钙、高蛋白和高钙十高蛋白饲料(试验组).试验第95天,各组随机取6只鸡采集血清,检测生化指标,随后扑杀,采集肝脏组织检测黄嘌呤氧化酶活性(XOD)、白细胞介素1(IL-1)、白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α),并将肝脏包埋切片进行HE和油红染色.[结果]各试验组与对照组相比,血清尿酸水平和肝中XOD活性升高;天门冬氨酸氨基转移酶、丙氨酸氨基转移酶升高,2、4组的总蛋白、白蛋白和球蛋白升高;总胆红素、直接胆红素和间接胆红素降低;高密度脂蛋白、低密度脂蛋白和胆固醇降低,甘油三酯升高;肝中IL-1、IL-6和TNF-α升高,而高密度脂蛋白、低密度脂蛋白、胆固醇、甘油三酯与对照组相近.显微镜下观察发现:肝内有坏死灶和炎性细胞浸润,肝细胞索排列紊乱,肝细胞空泡变性,少数鸡肝细胞内有脂肪滴.[结论]饲喂高钙和/或高蛋白饲料使鸡肝的结构和功能不同程度受损,高钙+高蛋白组受损更为严重.