發(fā)布時(shí)間：2018-09-18 06:52

【摘要】：人體內(nèi)有許多致病因素會(huì)影響腸道正常菌群的生長(zhǎng),從而導(dǎo)致其失調(diào),出現(xiàn)諸如腹痛、腹瀉等臨床癥狀。微生態(tài)學(xué)研究表明益生菌能粘附腸上皮細(xì)胞,形成天然生物屏障,顯著降低這類疾病的發(fā)生率。進(jìn)入腸道內(nèi)的益生菌通過粘附定植于腸粘膜表面以保護(hù)腸粘膜上皮細(xì)胞免受各種病原微生物的損傷,因此益生菌在腸道內(nèi)的粘附定植是其發(fā)揮生理作用的前提和基礎(chǔ)。目前,國(guó)內(nèi)外微生態(tài)制劑的評(píng)估方法有很多,但是缺乏橫向比較的資料。因此,本研究試圖對(duì)比益生菌粘附能力評(píng)估模型,以期提供這方面的基礎(chǔ)資料。 方法:以培菲康產(chǎn)品菌為模示菌株,采用革蘭氏染色、ELISA、放射性同位素標(biāo)記法和粘附抑制四種模型,分別評(píng)估了嗜酸乳桿菌、雙歧桿菌、糞腸球菌的粘附能力。通過靈敏度、時(shí)間、費(fèi)用方面的對(duì)比,確定最佳的評(píng)估模型。采用氯化鋰對(duì)乳桿菌的S層蛋白進(jìn)行提取,觀察表面蛋白(S層蛋白)對(duì)乳桿菌粘附能力評(píng)估模型的影響。 結(jié)果:益生菌的粘附性能檢測(cè)結(jié)果表明,三種粘附模型對(duì)益生菌粘附能力的評(píng)估基本一致,革蘭氏染色模型相對(duì)其他模型更為直觀、省時(shí),而且也不需要價(jià)格昂貴的分析儀器,可以作為益生菌粘附能力評(píng)價(jià)標(biāo)準(zhǔn)模型。益生菌的表面(S層)蛋白被抽提以后,其粘附能力被顯著影響(P0.01)。因此采用粘附模型評(píng)估微生態(tài)制劑應(yīng)盡量避免化學(xué)試劑對(duì)S層蛋白的影響。 結(jié)論:革蘭氏染色模型可以用做標(biāo)準(zhǔn)益生菌的粘附能力評(píng)估模型,具有靈敏度高,操作簡(jiǎn)易等特點(diǎn)。益生茵可抑制致病菌對(duì)腸上皮樣細(xì)胞HT-29的粘附和侵襲,減弱致病菌對(duì)腸上皮細(xì)胞樣HT-29的損傷。
[Abstract]:There are many pathogenic factors that affect the growth of normal intestinal flora, leading to its disorders, such as abdominal pain, diarrhea and other clinical symptoms. Microecological studies showed that probiotics could adhere to intestinal epithelial cells and form natural biological barrier, which significantly reduced the incidence of these diseases. Probiotics entering the intestinal tract can protect the intestinal epithelial cells from the damage of various pathogenic microorganisms by adhering to the intestinal mucosal surface, so the adhesion and colonization of probiotics in the intestinal tract is the premise and basis of its physiological function. At present, there are many evaluation methods for microecological agents at home and abroad, but lack of horizontal comparison data. Therefore, this study attempts to compare the evaluation model of probiotic adhesion in order to provide basic data. Methods: the adhesiveness of Lactobacillus acidophilus, Bifidobacterium faecalis and Enterococcus faecalis were evaluated by Gram-stained ELISA, radioisotope labeling and adhesion inhibition. Determine the best evaluation model by comparing sensitivity, time, and cost. The S-layer protein of Lactobacillus was extracted by lithium chloride, and the influence of surface protein (S-layer protein) on the adhesion ability of Lactobacillus was observed. Results: the results of adhesion test of probiotics showed that the three adhesion models were basically consistent in evaluating the adhesion ability of probiotics, and Gram staining model was more intuitive, time-saving, and did not need expensive analytical instruments compared with other models. It can be used as the standard model for evaluating the adhesion ability of probiotics. The adhesion of probiotic surface protein (S layer) was significantly affected (P0.01). Therefore, the effect of chemical reagent on S layer protein should be avoided by using adhesion model to evaluate microecological preparation. Conclusion: Gram-staining model can be used to evaluate the adhesion ability of standard probiotics with high sensitivity and easy operation. Probiotics could inhibit the adhesion and invasion of pathogenic bacteria to intestinal epithelial-like cells (HT-29) and attenuate the damage of pathogenic bacteria to intestinal epithelial-like HT-29.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R371

【引證文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 許飛利;我國(guó)食品工業(yè)常用益生乳酸菌菌種分型與溯源數(shù)據(jù)庫(kù)的研究[D];江南大學(xué);2011年

相關(guān)碩士學(xué)位論文 前1條

1 鄒芳;雞源乳酸菌對(duì)肉雞腸道粘蛋白的粘附性研究[D];東北農(nóng)業(yè)大學(xué);2010年

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本文編號(hào)：2247119