本文選題：乳酸菌 + 免疫調(diào)節(jié)��； 參考：《揚州大學》2016年碩士論文

【摘要】：乳酸菌是人體腸道中重要的有益菌,可以與腸道粘膜免疫系統(tǒng)相互作用,在抑制腸道致病菌、調(diào)節(jié)機體免疫功能、維持腸道微生態(tài)平衡等方面發(fā)揮了重要作用。當腸道菌群的平衡受到破壞時,宿主的正常代謝就會受到擾亂,甚至導致疾病。本文對源于廣西巴馬和新疆和田長壽人群糞便中的乳酸菌進行體外分離篩選,研究其在體內(nèi)的免疫調(diào)節(jié)作用,并利用實時熒光定量PCR技術(shù)和核磁共振技術(shù)探討其對免疫低下大鼠腸道菌群及宿主代謝的影響,從而為益生菌干預亞健康膳食提供助益。(1)從廣西巴馬和新疆和田長壽人群糞便中分離乳酸菌,并研究其體外疏水能力、對細胞黏附能力、耐酸耐膽鹽能力及抑菌特性和耐藥性。結(jié)果表明,分離獲得的153株乳酸菌中,10株乳酸菌的表面疏水能力較好、對細胞黏附能力較強；進一步研究發(fā)現(xiàn),10株乳酸菌中菌株P(guān)W1、LV108、XJH301在pH2.0的人工胃液中培養(yǎng)2h后的存活率分別為34.08%、61.35%、47.15%,在人工腸液中培養(yǎng)4h后的存活率分別為61.42%、86.03%、55.03%,在濃度0.3%的膽鹽MRS培養(yǎng)基中培養(yǎng)24h后的存活率分別為49.36%、45.24%、37.19%,顯著高于其它菌株(P0.05)；對大腸桿菌的抑菌圈分別為16.76mm、20.49mm、14.35mm,對沙門氏菌的抑菌圈分別為13.23mm、17.19mm、10.63mm,對金黃色葡萄球菌的抑菌圈分別為1176mm、14.01mm、10.10mm;菌株LV108對利福平、四環(huán)素、頭孢唑啉、氯霉素、頭孢拉定五種抗生素均敏感,對鏈霉素中度敏感,耐藥性低于其他菌株；經(jīng)API及16SrDNA測序?qū)V108菌株鑒定為鼠李糖乳桿菌(Lactobacillus rhamnosus)。(2)以左旋咪唑作為陽性對照,通過連續(xù)3天腹腔注射40mg/kg環(huán)磷酰胺的方法建立免疫低下大鼠模型。在建模期間,大鼠體重增加緩慢,胸腺生長受到抑制。血清中的IL-2、IgG、IgA含量、溶血素含量顯著降低(P0.05)；大鼠肝、脾巨噬細胞的廓清指數(shù)K、吞噬指數(shù)α顯著低于對照組(P0.05),脾淋巴細胞數(shù)量顯著下降(P0.05),建模成功。干預4周后發(fā)現(xiàn),L. rhamnosus LV108菌體懸浮液及其發(fā)酵乳能顯著提高大鼠脾臟指數(shù)和胸腺指數(shù)(P0.05)；同時還能顯著增加大鼠血清中IL-2、ⅠgG含量(P0.05)；刺激B淋巴細胞分化增殖,提高血清中的溶血素含量；顯著提高免疫低下大鼠的肝、脾巨噬細胞廓清指數(shù)K、吞噬指數(shù)α(P0.05),提高了大鼠單核巨噬細胞系統(tǒng)功能；促進大鼠脾淋巴細胞增殖,進而參與機體脾淋巴細胞的免疫應(yīng)答。(3)利用實時熒光定量PCR法研究大鼠腸道菌群的變化。結(jié)果表明,與對照組相比,免疫低下大鼠腸道菌群發(fā)生了變化,其中乳酸菌和雙歧桿菌的含量顯著降低(P0.05)而腸桿菌和腸球菌的含量顯著增加(P0.05)。干預4周后發(fā)現(xiàn),L. rhamnosus LV108菌體懸浮液及其發(fā)酵乳能顯著降低大鼠腸道中腸桿菌、腸球菌的含量(P0.05),而顯著增加大鼠腸道中乳酸菌及雙歧桿菌含量(P0.05)。L. rhamnosus LV108發(fā)酵乳對大鼠腸道菌群的改善幅度要大于L. rhamnosus LV108菌懸液,表明活的LV108菌體既能通過自身發(fā)揮作用,還能通過其代謝產(chǎn)物來改善腸道菌群。相關(guān)性分析發(fā)現(xiàn),大鼠腸道內(nèi)乳酸菌的含量同其血清中TNF-a及ⅠgA的水平呈顯著正相關(guān)(P0.05)。(4)利用核磁共振技術(shù)研究大鼠腸道代謝產(chǎn)物的變化。結(jié)果發(fā)現(xiàn),與對照組相比,免疫低下大鼠糞便代謝產(chǎn)物中異亮氨酸、丙氨酸、丁酸、琥珀酸、磷酸肌酸、氧化三甲胺、牛磺酸、β-葡萄糖、α-葡萄糖、尿嘧啶等成分含量降低。L. rhamnosus LV108菌體懸浮液及發(fā)酵乳對大鼠進行4周干預后,增加了大鼠糞便代謝產(chǎn)物中異亮氨酸、丙氨酸、賴氨酸等氨基酸的含量,改善了大鼠的氨基酸代謝和能量代謝；增加了乳酸、琥珀酸等有機酸的含量,抑制了腸道中致病菌的生長；增加了α-葡萄糖、N-乙酰糖蛋白、TMA、β-葡萄糖等成分的含量,改善了機體腸道菌群的代謝。
[Abstract]:Lactic acid bacteria, an important beneficial bacteria in the intestinal tract, can interact with the intestinal mucosal immune system. It plays an important role in inhibiting the intestinal pathogenic bacteria, regulating the immune function of the body and maintaining the intestinal microecological balance. When the balance of the intestinal flora is destroyed, the normal metabolism of the host will be disturbed and even causes the disease. In vitro isolation and screening of lactic acid bacteria derived from the feces of Guangxi Bama and Xinjiang Hetian long - lived population, the immunomodulatory effect of the lactic acid bacteria in the body was studied in this paper. The effects on intestinal microflora and host metabolites of immunocompromised rats were investigated by real time fluorescence quantitative PCR technique and nuclear magnetic resonance technique, so as to interfere with subhealth for probiotics. (1) isolation of lactic acid bacteria from the feces of Guangxi Bama and Xinjiang Hetian long - lived population, and study the hydrophobic ability in vitro, cell adhesion, acid resistance, salt tolerance and antimicrobial resistance and resistance. The results showed that of the 153 Lactobacillus Isolated, 10 Lactobacillus strains had better surface hydrophobicity and cell adhesion energy. The survival rate of the 10 strains of lactic acid bacteria PW1, LV108 and XJH301 in the artificial gastric juice of pH2.0 was 34.08%, 61.35%, 47.15% respectively. The survival rate of 4H in the artificial intestinal juice was 61.42%, 86.03%, 55.03%, respectively, and the survival rate of 24h in the bile salt MRS culture medium with the concentration of 0.3% was 49.36%, respectively. 45.24%, 37.19%, significantly higher than the other strains (P0.05), the Bacteriostasis of Escherichia coli were 16.76mm, 20.49mm, 14.35mm, and the bacteriostatic ring of Salmonella was 13.23mm, 17.19mm, 10.63mm, respectively, 1176mm, 14.01mm, 10.10mm for Staphylococcus aureus, and LV108 against Li Fuping, tetracycline, cefazolin, chloramphenicol, head. Five kinds of antibiotics were sensitive, and streptomycin was moderately sensitive and drug resistance was lower than that of other strains. LV108 was identified as Lactobacillus rhamnosus (Lactobacillus rhamnosus) by API and 16SrDNA sequencing. (2) levamisole was used as a positive control, and the immunocompromised rat model was established by intraperitoneal injection of 40mg/kg cyclophosphamide for 3 days. During the modeling period, the body weight increased slowly and the thymus growth was inhibited. The content of IL-2, IgG, IgA in the serum, the content of the hemolysin decreased significantly (P0.05), the clearance index of the liver and spleen macrophages in rats was K, the phagocytic index alpha was significantly lower than that of the control group (P0.05), the number of spleen lymphocytic cells decreased significantly (P0.05), and the modeling was successful. After 4 weeks of intervention, L. was found, L. Rhamnosus LV108 mycelium suspension and its fermented milk could significantly increase the spleen index and thymus index (P0.05) in rats, and also significantly increase the serum IL-2, gG content (P0.05), stimulate the proliferation of B lymphocytes and increase the content of hemolysin in the serum, and significantly improve the liver and spleen macrophage clearance finger in the immunocompromised rats. K, phagocytic index alpha (P0.05), increased the function of mononuclear macrophage system in rats, promoted the proliferation of spleen lymphocyte in rats, and then participated in the immune response of the spleen lymphocyte of the body. (3) the changes of intestinal flora in rats were studied by real-time fluorescence quantitative PCR. The results showed that the intestinal flora of immunocompromised rats was changed compared with the control group. The content of lactic acid bacteria and bifidobacteria significantly decreased (P0.05) and the content of enterobacteria and Enterococcus increased significantly (P0.05). After 4 weeks of intervention, it was found that L. rhamnosus LV108 mycelium suspension and its fermented milk could significantly reduce intestinal bacilli and Enterococcus (P0.05) in rats, and significantly increase the lactic acid bacteria and bifidobacteria in the intestine of rats. The improvement of intestinal microflora by bacilli content (P0.05).L. rhamnosus LV108 is greater than that of L. rhamnosus LV108 bacteria suspension. It shows that the living LV108 bacteria can not only play its role, but also improve the intestinal microflora through its metabolites. The correlation analysis shows that the content of lactic acid bacteria in the intestine of rats is the same as that of the serum in the rat. There was a significant positive correlation with the level of I gA (P0.05). (4) the changes in intestinal metabolites of rats were studied by NMR. Results showed that compared with the control group, isoleucine, alanine, butyric acid, succinic acid, creatine phosphocreatine, trimethylamine, taurine, beta glucose, alpha glucose, uracil, and so on were compared with the control group. The content of.L. rhamnosus LV108 bacterial suspension and fermented milk was reduced for 4 weeks. The contents of isoleucine, alanine, lysine and other amino acids were increased in rats' fecal metabolites, which improved the amino acid metabolism and energy metabolism in rats, increased the content of lactic acid, succinic acid and other organic acids, and inhibited the intestinal tract. The growth of pathogenic bacteria increased the content of alpha glucose, N- acetylglycoprotein, TMA, beta glucose and other components, and improved the metabolism of intestinal flora.

【學位授予單位】：揚州大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R151

【參考文獻】

相關(guān)期刊論文 前10條

1 陳超;鄭成中;;腸道菌群對人體免疫及兒童過敏性疾病的作用[J];國際兒科學雜志;2013年01期

2 胡旭;王濤;王瀝;金鋒;;腸道共生微生物與健康和疾病[J];中國微生態(tài)學雜志;2012年12期

3 呂穎堅;黃俊明;蔡玟;黃瓊;黃建康;李文立;陳壁鋒;胡帥爾;;環(huán)磷酰胺和氫化可的松建立免疫抑制模型方法比較[J];衛(wèi)生研究;2012年06期

4 楊會玲;高玉鵬;周利勇;許堯興;李艷麗;許少春;;合生素對肉仔雞腸道微生物區(qū)系的影響[J];中國農(nóng)業(yè)科學;2011年23期

5 徐仁應(yīng);萬燕萍;方啟宇;陸薇;張洪波;蔡威;;高脂飲食對大鼠腸道益生菌和肝脂肪含量的影響[J];營養(yǎng)學報;2011年04期

6 陳帥;周方方;王蔭榆;郭本恒;;干酪乳桿菌LC2W對胃上皮細胞的粘附性質(zhì)及其與菌體表面性質(zhì)的關(guān)系[J];中國微生態(tài)學雜志;2010年11期

7 徐麗丹;鄒積宏;文姝;李華軍;袁杰利;;降血壓乳酸菌發(fā)酵乳對原發(fā)性高血壓大鼠的降壓效果[J];中國微生態(tài)學雜志;2010年10期

8 宋園亮;李海燕;柳陳堅;;乳酸菌治療乳糖不耐受癥的研究進展[J];中國微生態(tài)學雜志;2010年08期

9 王春敏;李麗秋;;人體腸道正常菌群的研究進展[J];中國微生態(tài)學雜志;2010年08期

10 高權(quán)新;吳天星;王進波;;腸道微生物與寄主的共生關(guān)系研究進展[J];動物營養(yǎng)學報;2010年03期

，

本文編號：1820262