瑞士乳杆菌对小鼠肠道粘膜免疫的影响

发布时间：2018-04-19 18:56

  本文选题：瑞士乳杆菌 + 肠道微生物区系　； 参考：《天津商业大学》2008年硕士论文

【摘要】： 益生菌作为一种活的有机体,存在于肠道粘膜上,并对肠道黏膜有保护和调节作用,可以治疗各种肠道疾病。益生菌可直接作用于宿主的免疫系统,诱发肠道免疫,并刺激胸腺、脾脏和法氏囊等免疫器官的发育,促进巨噬细胞活力或发挥佐剂作用,活化肠粘膜内相关淋巴组织,使SIgA分泌增加。益生菌在体液免疫和细胞免疫中发挥着调节作用,其保健作用的机理成为近年来研究的重点内容。本文选用瑞士乳杆菌(Lactobacillus helveticusTS206)作为研究菌株,系统地研究了其对小鼠肠道粘膜免疫的影响。本研究首先采用了改良培养基的方法,选择了瑞士乳杆菌的最适培养基;接着利用蛋白质的溶解特性和梯度离心的方法建立了菌体细胞分离纯化技术;然后采用选择性培养技术系统研究了小鼠肠道微生物区系;用放射性免疫(RIA)和酶联免疫(ELISA)检测技术系统评价了小鼠肠道粘膜中SIgA和肠道粘膜中的分泌细胞因子;同时观察条件致病菌(E.coli)灌胃组对小鼠粘膜免疫的影响,研究瑞士乳杆菌对小鼠肠粘膜免疫系统的作用方式。 实验结果表明:①瑞士乳杆菌的最适培养基为溴甲酚绿指示剂培养基。②瑞士乳杆菌对小鼠肠道微生物区系调节的最佳灌胃剂量为2×10~7cfu/d。③长期灌胃瑞士乳杆菌对小鼠肠道粘膜中SIgA含量的影响:瑞士乳杆菌灌胃组小鼠肠道粘膜中SIgA的含量在第10天达到最大值,和对照组相比差异极显著(P＜0.01),肠道粘膜中SIgA含量显著升高。短期灌胃瑞士乳杆菌对小鼠肠道粘膜中SIgA含量的影响:瑞士乳杆菌灌胃组小鼠肠道粘膜中SIgA含量在24h内持续上升。④长期灌胃瑞士乳杆菌对小鼠肠道中细胞因子的影响:瑞士乳杆菌灌胃组小鼠肠道中四种细胞因子和比对照组明显升高(P＜0.05),瑞士乳杆菌灌胃组小鼠肠道内Th1型细胞因子(IL-2,IFN-γ)在第3d前达到最大值;Th2型细胞因子(IL-4,IL-6)在第5d达到顶峰;Th1型和Th2型T细胞因子的比例和对照组相比无显著性差异(P＞0.05),肠道免疫功能未出现异常;大肠杆菌灌胃组则出现了异常。大肠杆菌灌胃组则出现异常。短期灌胃瑞士乳杆菌对小鼠肠道中细胞因子的影响:IL-2和IL-6在第6h达到最高点;IL-4在第1h达到最高点;IFN-γ在第3h达到最高值;Th2型细胞因子比Th1型的多,于是可以推测出Th1型T细胞可能是比Th2型的少一些,这是由于T细胞短时间内不同分化方向造成的。和长期灌胃组相比短期灌胃组细胞因子的波动比较大,而长期灌胃组的结果更能反映瑞士乳杆菌对小鼠肠道粘膜免疫的调理作用。
[Abstract]:Probiotics, as a living organism, exist on the intestinal mucosa and can protect and regulate the intestinal mucosa, which can treat various intestinal diseases. Probiotics can directly act on the host's immune system, induce intestinal immunity, stimulate the development of immune organs such as thymus, spleen and bursa of Fabricius, promote the activity of macrophages or play an adjuvant role, and activate the lymphoid tissues associated with intestinal mucosa. Increase the secretion of SIgA. Probiotics play a regulatory role in humoral and cellular immunity. The effects of Lactobacillus helveticus TS206 on intestinal mucosal immunity in mice were studied systematically. In this study, the method of improved culture medium was used to select the optimum medium for Lactobacillus helicus, and then the isolation and purification technology of bacteria somatic cells was established by using protein dissolution characteristics and gradient centrifugation. The intestinal microflora of mice was studied by selective culture technique, and the SIgA and secreted cytokines in intestinal mucosa of mice were evaluated by radioimmunoassay (RIA) and enzyme linked immunosorbent assay (Elisa). At the same time, the effects of E. coli on mucosal immunity in mice were observed, and the effects of Lactobacillus helicum on intestinal mucosal immune system in mice were studied. The results showed that the optimal medium for Lactobacillus Switzerland was bromocresol green indicator medium. 2 Lactobacillus helittidis for the regulation of intestinal microflora in mice. 2 脳 10~7cfu/d.3 for a long period of time. Effect of SIgA content in intestinal mucosa of mice: the content of SIgA in intestinal mucosa of Lactobacillus helicum group reached its maximum on the 10th day. Compared with the control group, the difference was very significant (P < 0.01), and the content of SIgA in intestinal mucosa was significantly higher than that in the control group (P < 0.01). Effect of Lactobacillus helicum on SIgA content in intestinal mucosa of mice: the content of SIgA in intestinal mucosa of Lactobacillus Switzerland group increased continuously within 24 hours. 4. Long term administration of Lactobacillus helicus on cytokines in intestinal tract of mice The effects of lactobacillus helicobacterium on the four cytokines in the intestinal tract of mice were significantly higher than those in the control group (P < 0.05), and the Th1 type cytokine (IL-2IFN- 纬) in the intestinal tract of the Lactobacillus hominis group reached the maximum value before the third day, and the level of IL-4 / IL-6 of Th2 cytokines was significantly higher than that of the control group (P < 0.05). On the 5th day, there was no significant difference in the proportion of T cytokines of Th1 type and Th2 type between the control group and the control group (P > 0.05), and there was no abnormal intestinal immune function. The group of Escherichia coli was abnormal. The group of Escherichia coli was abnormal. Effects of short term administration of Lactobacillus helicum on cytokines in the intestinal tract of mice, the highest levels of IL 4 and IL 2 were reached at 6 h and the highest level of IFN- 纬 at 1 h. Th2 type cytokines were more than those of Th1 type at 3 h. It can be inferred that Th1 type T cells may be less than those of Th2 type, which is due to the different differentiation directions of T cells in a short period of time. Compared with the long-term group, the fluctuation of cytokines in the short-term group was larger than that in the long-term group, and the results of the long-term group could reflect the regulation effect of Lactobacillus helicum on the intestinal mucosal immunity of mice.
【学位授予单位】：天津商业大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R371

【引证文献】

相关博士学位论文 前1条

1 刘海燕;转nisI基因植物乳杆菌Lactobacillus plantarum 590对SD大鼠肠道健康的影响研究[D];江南大学;2011年

相关硕士学位论文 前1条

1 李美君;乳铁蛋白对早期断奶仔猪肠道结构和免疫功能的影响研究[D];湖南农业大学;2011年

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