發(fā)布時間：2018-07-07 13:58

  本文選題：植物乳桿菌 + 生物吸附　； 參考：《江南大學(xué)》2014年碩士論文

【摘要】：環(huán)境污染致使大量的銅進(jìn)入食物鏈中，并不斷富集，嚴(yán)重威脅食品安全及人體健康。過量的銅會引發(fā)機(jī)體氧化應(yīng)激反應(yīng)和損傷，導(dǎo)致肝臟病變、腦部受損，出現(xiàn)肝腫大、神經(jīng)衰弱、智力下降等癥狀。有研究表明，乳酸菌能夠有效吸附銅離子，且乳酸菌具有抗氧化、消除人體自由基等免疫調(diào)節(jié)作用，此外使用成本低、屬于食品安全級（GRAS）微生物等特點(diǎn)都是其優(yōu)勢所在。因此，探究膳食攝入益生乳酸菌是否能夠緩解重金屬銅毒性是一個非常具有研究意義的課題。本論文以乳酸菌作為研究對象，以對銅離子的吸附能力及耐受能力作為指標(biāo)進(jìn)行篩選，獲得了一株對銅具有較強(qiáng)吸附能力及耐受能力的植物乳桿菌，，并對其進(jìn)行了動物實(shí)驗的功效驗證及實(shí)際應(yīng)用價值的開發(fā)。 針對菌株對銅的吸附和耐受特性，選取16株乳酸菌進(jìn)行體外篩選，得到一株能夠高效吸附銅離子并且生長不受高濃度銅抑制的理想菌株植物乳桿菌CCFM8246，其最大吸附量達(dá)到19.7mg/g，在含銅離子濃度為50mg/L，150mg/L、500mg/L的MRS培養(yǎng)基中能夠正常生長。 在植物乳桿菌CCFM8246的體內(nèi)實(shí)驗功能評價中，通過灌胃小鼠銅溶液（濃度分別為100mg/kg，150mg/kg）成功建立小鼠銅暴露損傷模型，每日灌喂植物乳桿菌CCFM8246（2.0×1010CFU/mL，0.2mL）加以干預(yù)和治療。每周收集糞便并記錄體重等生理指標(biāo)，通過水迷宮實(shí)驗研究銅暴露對小鼠智力受損情況的影響及植物乳桿菌CCFM8246對相關(guān)損傷的緩解作用。處死小鼠后收集血液、肝臟、腎臟、腦等進(jìn)行生理生化分析，比較組織和糞便中的銅含量、血清及組織勻漿的氧化應(yīng)激相關(guān)指標(biāo)。結(jié)果表明，與銅暴露模型組相比，植物乳桿菌CCFM8246灌胃組小鼠糞便中的銅含量顯著升高，最高可達(dá)到6倍以上，說明植物乳桿菌CCFM8246能夠有效降低小鼠腸道對銅的吸收。與之對應(yīng)，植物乳桿菌CCFM8246灌胃組小鼠組織中銅含量明顯降低，在最顯著實(shí)驗組中銅含量減少量可達(dá)到75.9%，進(jìn)一步驗證了其排銅作用。同時，植物乳桿菌CCFM8246對銅暴露毒性的兩大重要器官肝臟和腦部也起到了一定的保護(hù)作用。植物乳桿菌CCFM8246灌胃組小鼠肝臟勻漿中谷胱甘肽過氧化物酶（GSH-PX）、超氧化物歧化酶（SOD）活性升高，而丙二醛（MDA）含量顯著降低，表明植物乳桿菌CCFM8246能有效緩解銅暴露對肝臟造成的氧化損傷。血清中谷丙轉(zhuǎn)氨酶（ALT）、谷草轉(zhuǎn)氨酶（AST）活性降低，再次驗證了植物乳桿菌CCFM8246對肝臟損傷的緩解作用。水迷宮實(shí)驗也證實(shí)植物乳桿菌CCFM8246能夠有效緩解銅暴露導(dǎo)致的小鼠智力損傷，提高學(xué)習(xí)記憶能力。本研究結(jié)果顯示，植物乳桿菌CCFM8246確實(shí)具有緩解小鼠銅毒性作用，可以作為一個新的飲食治療方法治療銅誘導(dǎo)損傷。 植物乳桿菌CCFM8246在豆乳制品中的應(yīng)用研究表明，該菌株在豆乳中生長情況良好，感官品質(zhì)較佳。由此可見，植物乳桿菌CCFM8246具有發(fā)酵豆乳制品的潛力與優(yōu)勢，可用于相關(guān)產(chǎn)品的研究開發(fā)和商業(yè)化生產(chǎn)。
[Abstract]:Environmental pollution causes a large amount of copper into the food chain and enriching, which seriously threatens food safety and human health. Excessive copper can cause oxidative stress reaction and damage to the body, causing liver disease, brain damage, hepatomegaly, neurasthenia, mental decline and so on. Studies have shown that lactic acid bacteria can effectively adsorb copper ions, and Lactic acid bacteria have the antioxidation, eliminate the immune regulation of human free radicals and so on. In addition, it has the advantages of low cost, which belong to the food safety grade (GRAS) microorganism. Therefore, it is a very important task to explore whether the dietary intake of probiotic lactic acid bacteria can alleviate the toxicity of heavy metal copper. As an object of study, a plant lactobacillus with strong adsorption capacity and tolerance to copper was obtained by screening the adsorption capacity and tolerance capacity of copper ions, and the efficacy of animal experiments and the development of practical application value were also carried out.
In view of the adsorption and tolerance characteristics of the strain to copper, 16 strains of lactic acid bacteria were selected in vitro to obtain an ideal strain of Lactobacillus plantarum CCFM8246, which could efficiently adsorb copper ions and grow without high concentration of copper. The maximum adsorption capacity reached 19.7mg/g. In the MRS medium containing the concentration of copper ion concentration of 50mg/L, 150mg/L and 500mg/L Normal growth.
In the evaluation of the experimental function of Lactobacillus plantarum CCFM8246 in vivo, the mice copper exposure damage model was established by gavage of mice copper solution (100mg/kg, 150mg/kg). The mice were fed with Lactobacillus plantarum CCFM8246 (2 x 1010CFU/mL, 0.2mL) daily to intervene and treat them. The effect of copper exposure on mental impairment in mice and the alleviating effect of Lactobacillus plantarum CCFM8246 on related damage in mice was studied. The blood, liver, kidney and brain were collected and the copper content in tissue and feces were compared, and the related indexes of oxidative stress in serum and tissue homogenate were compared. Compared with the exposed model group, the copper content in the feces of the mice treated with Lactobacillus plantarum CCFM8246 was significantly higher than that of 6 times, indicating that Lactobacillus plantarum CCFM8246 could effectively reduce the absorption of copper in the intestinal tract of mice. The copper content in the mouse group of Lactobacillus plantarum CCFM8246 was significantly reduced and the most significant experimental group was in the most significant experimental group. The reduction of copper content could reach 75.9%, which further verified the effect of copper excretion. At the same time, Lactobacillus plantarum CCFM8246 also played a protective role in the liver and brain of two important organs of copper exposure toxicity. The glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD) in the liver homogenate of mice treated with Lactobacillus plantarum The activity increased and the content of malondialdehyde (MDA) decreased significantly, indicating that Lactobacillus plantarum CCFM8246 could effectively alleviate the oxidative damage caused by copper exposure to the liver. The activity of cereal alanine aminotransferase (ALT) and glutarine transaminase (AST) in serum was reduced. The remission of Lactobacillus plantarum CCFM8246 to liver injury was verified. The water maze test also confirmed the plant milk. Bacilli CCFM8246 can effectively alleviate the mental impairment in mice caused by copper exposure and improve the learning and memory ability. The results of this study showed that Lactobacillus plantarum CCFM8246 did alleviate copper toxicity in mice and could be used as a new dietary therapy to treat copper induced injury.
The application of Lactobacillus plantarum CCFM8246 in soybean milk products showed that the strain grew well in soybean milk and had better sensory quality. Thus, Lactobacillus plantarum CCFM8246 had the potential and advantage of fermented soybean milk products, and could be used in the research and development of related products and commercial production.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R965

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