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實(shí)時(shí)熒光定量PCR檢測(cè)小鼠腸道乳酸菌及細(xì)胞因子含量的研究

發(fā)布時(shí)間:2018-04-24 08:34

  本文選題:乳酸菌 + 實(shí)時(shí)熒光定量PCR ; 參考:《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2015年碩士論文


【摘要】:本實(shí)驗(yàn)用酸奶、駝奶、仔豬糞便、雞腸道分離的6株乳酸菌的活菌體制成菌懸液灌服小鼠,一日一次,連續(xù)灌服30 d,并分別在第15、30 d和停灌后7d每組抽出10只小鼠處死立即取腸道及腸道內(nèi)容物和脾臟,然后提腸道及腸道內(nèi)容物的DNA和脾臟總RNA后用實(shí)時(shí)熒光定量PCR檢測(cè)小鼠腸道及腸道內(nèi)容物乳酸菌數(shù)量和小鼠脾臟中細(xì)胞因子IL-2、IL-4、IL-6、TNF-α、IFN-γ表達(dá)量變化的研究。結(jié)果表明,6株乳酸菌能夠定植小鼠腸道內(nèi),但不同來源乳酸菌的定植能力不一樣,跟對(duì)照組比較:飼喂第15d時(shí)LT組極顯著增多(P0.01),其次LZ1組顯著增多(P0.05),其余乳酸菌與對(duì)照組沒有顯著差異;第30d時(shí)LT、LZ1組顯著增多(P0.05),其余乳酸菌沒有顯著差異;停喂后7d LZ1組顯著增多(P0.05),其余乳酸菌沒有顯著差異。對(duì)于飼喂的天數(shù),連續(xù)飼喂乳酸菌可以保持小鼠腸道內(nèi)一定量的乳酸菌,一旦停止飼喂乳酸菌,小鼠腸道中定植的乳酸菌含量逐漸的減少。六株乳酸菌跟對(duì)照組比較:飼喂第15、30d時(shí)LJ2組極顯著提高了小鼠脾臟中IL-2、IL-4、IL-6、TNF-α、IFN-γ的表達(dá)量(P0.01),其次LZ2組顯著增加(P0.05),其余乳酸菌與對(duì)照組沒有顯著差異:停喂后7dLS、LZ2組顯著增加了小鼠脾臟中IL-2、IL-4、IL-6、TNF-α、IFN-γ的表達(dá)量(P0.05),其余乳酸菌與對(duì)照組沒有顯著差異。
[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- 偽 and IFN- 緯 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- 偽 -IFN- 緯 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- 偽 -IFN- 緯 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- 偽 and IFN- 緯 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.
【學(xué)位授予單位】:內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2015
【分類號(hào)】:S852.6

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