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大鼠派伊尔结内生Lactobacillus reuteri的分离及其调节sIgA产生的研究

发表时间:2016-08-15  浏览量:1260  下载量:315
全部作者: 王盼盼,李 亚,齐 策,施用晖,乐国伟,孙 进
作者单位: 江南大学食品学院,食品营养与功能因子研究中心;江南大学食品科学与技术国家重点实验室
摘 要: 目的:研究大鼠派伊尔结(Peyer’s patch,PP)内Lactobacillus reuteri对大鼠sIgA分泌的影响。方法:从PP分离乳杆菌,进行鉴定和分型分析,用分离的典型菌株(PP1和PP2)进行动物实验。将大鼠随机分为正常饲料组(CT)和维生素A缺乏饲料组(VAD),饲喂6周至VAD组形成维生素A缺乏症状,然后每组再分别分为三组,一组为对照,另两组分别灌胃PP1和PP2,即CT组、CT+PP1组、CT+PP2组和VAD组、VAD+PP1组、VAD+PP2组。第10周结束实验,用ELISA测定各组大鼠sIgA含量;免疫组化分析各组大鼠结肠黏膜IgA浆细胞分布;用末端限制性片段多态性(terminal restriction fragment polymorphism,T-RFLP)技术和乳杆菌特异性T-RFLP技术分析粪便菌群及PP和回肠内容物(ileum content,IC)乳杆菌构成。结果:大鼠PP分离的乳杆菌经16S rRNA基因测序鉴定均为L. reuteri,用扩增片段长度多态性(amplified fragment length polymorphism,AFLP)分型分为两个表型,代表菌株为PP1和PP2. 大鼠饲喂6周后VAD组粪便sIgA含量显著低于CT组(P<0.05),PP1和PP2都可以增加CT和VAD大鼠肠道菌群的β多样性,但仅使CT大鼠PP乳杆菌β多样性显著增加。粪便游离sIgA与PP乳杆菌多样性存在显著正相关关系(R2=0.795 8,P=0.011),也与多种PP内乳杆菌出现率显著正相关(R2=0.897 3,P=0.003),包括L. casei/L. paracasei、L. paraplantarum、L. acetotolerans、L. kefiranofaciens subsp. Kef.、L. curvatus和L. sakei. 结论:PP内L. reuteri促进sIgA分泌,该调节作用依赖于维生素A和PP乳杆菌多样性的改变。
关 键 词: 食品营养学;派伊尔结;维生素A缺乏;末端限制性片段多态性;肠道菌群
Title: Isolation of Lactobacillus reuteri from rat Peyer’s patches and its effect on sIgA production
Author: WANG Panpan, LI Ya, QI Ce, SHI Yonghui, LE Guowei, SUN Jin
Organization: Food Nutrition and Functional Factors Research Center, School of Food Science and Technology, Jiangnan University; State Key Laboratory of Food Science and Technology, Jiangnan University
Abstract: Objective: This study was focused on the effect of Peyer’s patch (PP) Lactobacillus reuteri in sIgA production of rats. Methods: SD rats were randomly divided into control group (CT) and vitamin A deficent group (VAD) and fed for 6 weeks. Lactobacilli were isolated from normal rat PP. All isolates were L. reuteri belong to two phenotypes according to amplified fragment length polymorphism (AFLP) analysis. Specific strain of each cluster, PP1 and PP2, were used to treat CT (CT+PP1, CT+PP2) and VAD (VAD+PP1, VAD+PP2) rats to 10th week, respectively. Fecal sIgA was determined with ELISA. sIgA distribution in colon mucosa was determined with immunohistochemistry. Terminal restriction fragment polymorphism (T-RFLP) and lactobacillus specific T-RFLP were used to analysis the fecal flora characteristics and lactobacillus differences between PP and ileum content (IC). Results: Fecal sIgA was significantly lower in VAD rats than control (P<0.05) after 6 weeks. Both PP1 and PP2 increased beta diversity of intestinal flora no matter in CT and VAD rats. They significantly increased beta diversity of PP Lactobacillus only in CT rats. Fecal free sIgA was correlated with PP Lactobacillus diversity (R2=0.795 8, P=0.011) and with the occurrence of L. casei/L. paracasei, L. paraplantarum, L. acetotolerans, L. kefiranofaciens subsp. Kef., L. curvatus and L. sakei (R2=0.897 3, P=0.003). Conclusion: Modulation of sIgA production by PP L. reuteri is dependent on vitamin A and change of Lactobacillus diversity in PP.
Key words: food nutriology; Peyer’s patch; vitamin A deficiency; terminal restriction fragment polymorphism; intestinal microflora
发表期数: 2016年8月第15期
引用格式: 王盼盼,李 亚,齐 策,等. 大鼠派伊尔结内生Lactobacillus reuteri的分离及其调节sIgA产生的研究[J]. 中国科技论文在线精品论文,2016,9(15):1603-1616.
 
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