实时荧光定量PCR检测小鼠肠道乳酸菌及细胞因子含量的研究

发布时间：2018-04-24 08:34

本文选题：乳酸菌 + 实时荧光定量PCR　；参考：《内蒙古农业大学》2015年硕士论文

【摘要】：本实验用酸奶、驼奶、仔猪粪便、鸡肠道分离的6株乳酸菌的活菌体制成菌悬液灌服小鼠,一日一次,连续灌服30 d,并分别在第15、30 d和停灌后7d每组抽出10只小鼠处死立即取肠道及肠道内容物和脾脏,然后提肠道及肠道内容物的DNA和脾脏总RNA后用实时荧光定量PCR检测小鼠肠道及肠道内容物乳酸菌数量和小鼠脾脏中细胞因子IL-2、IL-4、IL-6、TNF-α、IFN-γ表达量变化的研究。结果表明,6株乳酸菌能够定植小鼠肠道内,但不同来源乳酸菌的定植能力不一样,跟对照组比较：饲喂第15d时LT组极显著增多(P0.01),其次LZ1组显著增多(P0.05),其余乳酸菌与对照组没有显著差异；第30d时LT、LZ1组显著增多(P0.05),其余乳酸菌没有显著差异；停喂后7d LZ1组显著增多(P0.05),其余乳酸菌没有显著差异。对于饲喂的天数,连续饲喂乳酸菌可以保持小鼠肠道内一定量的乳酸菌,一旦停止饲喂乳酸菌,小鼠肠道中定植的乳酸菌含量逐渐的减少。六株乳酸菌跟对照组比较：饲喂第15、30d时LJ2组极显著提高了小鼠脾脏中IL-2、IL-4、IL-6、TNF-α、IFN-γ的表达量(P0.01),其次LZ2组显著增加(P0.05),其余乳酸菌与对照组没有显著差异：停喂后7dLS、LZ2组显著增加了小鼠脾脏中IL-2、IL-4、IL-6、TNF-α、IFN-γ的表达量(P0.05),其余乳酸菌与对照组没有显著差异。
[Abstract]:In this experiment, mice were administered with live bacteria suspension of 6 strains of lactic acid bacteria isolated from yogurt, camel milk, piglet feces and chicken intestines once a day. After 30 days of continuous administration, 10 mice in each group were killed at 1530 days and 7 days after stopping the administration, and the intestinal tract, intestinal contents and spleen were taken immediately. Then DNA of intestinal tract and intestinal contents and total RNA of spleen were extracted. The changes of lactic acid bacteria in intestinal tract and intestinal contents of mice and the expression of cytokine IL-2, IL-4, IL-4, TNF- 伪 and IFN- 纬 in the spleen of mice were detected by real-time fluorescence quantitative PCR. The results showed that 6 strains of lactic acid bacteria could colonize the intestinal tract of mice, but the colonization ability of lactic acid bacteria from different sources was different. Compared with the control group, the LT group significantly increased P0.01N on the 15th day, followed by the LZ1 group significantly increased the P0.05A, and the other lactic acid bacteria were not significantly different from the control group, and on the 30th day, the LZ1 group increased the P0.05N significantly, but the other lactic acid bacteria had no significant difference. In LZ1 group, there was a significant increase in P0.05, but there was no significant difference in other lactic acid bacteria. For the days of feeding, continuous feeding of lactic acid bacteria can keep a certain amount of lactic acid bacteria in the intestinal tract of mice. Once the feeding of lactic acid bacteria is stopped, the content of lactic acid bacteria in the intestinal tract of mice gradually decreases. Six strains of lactic acid bacteria compared with the control group: the LJ2 group significantly increased the expression of IL-2TNF- 伪 -IFN- 纬 in the spleen of mice on the 30th day after feeding, followed by the LZ2 group significantly increased the expression of P0.05, and the other lactic acid bacteria showed no significant difference between the control group and the control group: after 7 days of feeding, the LZ2 group significantly increased the expression of TNF- 伪 -IFN- 纬 in the spleen of mice, followed by the increase of P0.05 in the LZ2 group, and there was no significant difference between the other lactic acid bacteria and the control group. The expression of IL-2, IL-4, IL-6, TNF- 伪 and IFN- 纬 in the spleen of mice was compared with that of the control group (P 0.05). There was no significant difference between the other lactic acid bacteria and the control group.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.6

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