1
目 录
Abstract1
Key words1
引言2
1材料与方法2
1.1实验材料2
1.1.1 实验动物2
1.1.2试剂耗材2
1.1.3仪器设备2
1.2实验方法3
1.2.1饲养方法3
1.2.2切片制作3
1.2.2.1石蜡切片制作3
1.2.2.2冰冻切片制作3
1.2.3仔猪结肠、回肠HE染色3
1.2.4仔猪结肠、回肠免疫荧光染色3
1.2.5仔猪结肠、回肠TNFα及IL1β含量测定3
2结果与分析4
2.1仔猪肠黏膜形态学观察4
2.2杯状细胞数量变化5
2.3 TNFα及IL1β表达量比较 7
2.3.1仔猪结肠、回肠TNFα表达量7
2.3.2仔猪结肠、回肠IL1β表达量8
3讨论9
3.1 C30BS对仔猪肠绒毛发育的影响9
3.2 C30BS对仔猪肠道杯状细胞数量的影响9
3.3 C30BS对仔猪肠道细胞因子TNFα及IL1β表达量的影响9
4小结9
致谢9
参考文献10
重组表达类胡萝卜素的枯草芽孢杆菌
对仔猪肠黏膜的调控机制
动物药学专业学生 朱怡蕾
指导教师 庾庆华
摘要：C30类胡萝卜素是通过crtM和crtN两种类胡萝卜素合成相关酶，在枯草芽孢杆菌中合成的高纯度类胡萝卜素。本实验构建磷酸盐缓冲液（PBS）、枯草芽孢杆菌（Bacillus, BS）、表达C30类胡萝卜素的枯草芽孢杆菌（C30Bacillus, C30BS）三种饲料添加剂模型，分别对同批仔猪进行饲喂，一月后取仔猪肠组织。通过HE染色、免疫荧光染色及酶联免疫吸附试验等实验方法初步探明C30BS对仔猪肠道的作用，包括对肠黏膜形态结构、杯状细胞数量和分布的影响，对促炎细胞因子TNFα和IL1β的影响。进而推断C30BS对肠道黏膜免疫的调控作用。C30BS能促进肠道绒毛生长 *51今日免费论文网|www.jxszl.com +Q: ￥351916072￥ 
，增加隐窝深度，维护肠黏膜屏障的稳定；能增加肠隐窝内杯状细胞数量，利于肠道菌群的调控；饲喂后表达促炎细胞因子水平低，不会产生炎症反应。因此可以初步说明C30BS是一种安全的饲料添加剂。
Regulatory mechanism of Bacillus subtilis expressing carotenoids on intestinal mucosa of piglets
Student majoring in veterinary pharmacy Zhu Yilei
Tutor Yu Qinghua
Abstract：C30 carotenoids are highpurity carotenoids synthesized in Bacillus subtilis by two carotenoid synthetic enzymes called crtM and crtN. In this experiment, three kinds of feed additive models are built as PBS, Bacillus (BS) and C30Bacillus (C30BS) which were used to feed the same piglets, and the intestinal tissue of the piglets was taken in one month later. The effects of C30BS the intestinal tract of piglets, the structure of intestinal mucosa, the distribution of goblet cells, and effects on cell factors such as TNFα and IL1β were studied by HE staining, immunofluorescence staining and enzymelinked immunosorbent assay (ELISA). So as to deduce the regulation of intestinal mucosal immunity by C30BS. The C30BS can promote the growth of intestinal villi, and increase the crypt depth, make the intestinal mucosa presents a healthy state. It can increase the quantity of goblet cells within the intestinal crypt, which is advantageous to the intestinal flora regulation. The low expression levels of proinflammatory cytokines also shows that the C30BS is a safe type of feed additives which will not cause inflammatory reaction.

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