内蒙古乳制品中植物乳杆菌对小鼠肠道菌群及免疫功能的影响
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摘要
本研究对分离自内蒙古牧区乳及乳制品中的28株植物乳杆菌(Lactobacillus plantarum)进行了酸耐受性、胆盐耐受性和对细菌的抑菌性等生物学特性的研究,并对经以上试验筛选所得的L. plantarum KH13-1进行了质粒的检测和小鼠的肠道菌群及免疫调节的研究。
28株试验菌中,其中22株对酸和胆盐显示出一定的的耐受性,它们都能耐受0.3%的牛胆酸钠,且对pH3.0的生长环境表现出不同程度的耐受性。其中9株菌对酸的耐受性较强,存活率最高可达150.6%,其余菌株从23.4%～39.8%不等;这些菌株对大肠杆菌K99、金黄色葡萄球菌CMCC(B)26001、福氏志贺氏菌CMCC(B)sh51592、溶壁微球菌、枯草芽孢杆菌CMCC(B)63501等病原菌有广谱抑制作用,但对非病原菌的抑制作用较弱甚至没有,抑菌物质主要为有机酸和过氧化氢。L. plantarum KH13-1细胞中不含有质粒。
对体外试验筛选出的L. plantarum KH13-1进行了肠道菌群调节和增强机体免疫两种益生作用的研究。L. plantarum KH13-1具有调节抗生素所致菌群失调小鼠肠道菌群平衡的作用;摄入该菌后对免疫功能低下小鼠的免疫功能有增强作用,而对正常小鼠无显著影响;它的热致死菌体可促进小鼠脾细胞IL-4和IFN-γ的产生,但其活菌体对IL-4和IFN-γ的产生无影响。
In this study , the biological characteristic of 28 strains of Lactobacillus plantarum were analyzed, which isolated from traditional dairy products in pasture region of Inner Mongolia, such as tolerance to acid and bile salt, inhibition of their fermentation supernatant to the indicator . According to the screening experiment in vitro, L. plantarum KH13-1 was selected to a lot of studies, e.g checked the plasmid of L. plantarum KH13-1, its regulation function of immunity and intestinal flora of mice.
22 strains of L. plantarum showed resistance to acid and bile salt among of tested bacteria. All of them could grow in the medium contained 0.3 % sodium taurocholate, and show tolerance to acid pH3.0 in a certain degree. 9 strains of L. plantarum could resist acid well, survival rate can even reach 150.6%. The survival rate of other strains of bacteria was different from 23.4% to 39.8%; The 9 strains of L. plantarum had a wide antimicrobial activity to pathogenic bacteria, e.g Escherichia col, Staphylococcus aureus, Shigella flexneri, Micrococcus lysodeikticus, Bacillus cereus, et al. The antimicrobial activity was weak to nonpathogenic bacteria. The antimicrobial substance was mainly organic acid and. L. plantarum KH13-1 had no plasmid.
According to the screening experiment in vitro, L. plantarum KH13-1 was studied to regulation function of immunity and intestinal microflora of mice. L. plantarum KH13-1 could modulate the normal balance of intestinal microflora in mice with antibiotic-associated intestinal flora imbalance; And the results showed that the strain could enhance immune function of IDMM, but it had no effect on the specific immunity to normal mice, at the same time, its heat-killed bacteria could also induce IL-4 and IFN-γproduction by spleen cells from the mice, but live bacteria had no influence.
28株试验菌中,其中22株对酸和胆盐显示出一定的的耐受性,它们都能耐受0.3%的牛胆酸钠,且对pH3.0的生长环境表现出不同程度的耐受性。其中9株菌对酸的耐受性较强,存活率最高可达150.6%,其余菌株从23.4%～39.8%不等;这些菌株对大肠杆菌K99、金黄色葡萄球菌CMCC(B)26001、福氏志贺氏菌CMCC(B)sh51592、溶壁微球菌、枯草芽孢杆菌CMCC(B)63501等病原菌有广谱抑制作用,但对非病原菌的抑制作用较弱甚至没有,抑菌物质主要为有机酸和过氧化氢。L. plantarum KH13-1细胞中不含有质粒。
对体外试验筛选出的L. plantarum KH13-1进行了肠道菌群调节和增强机体免疫两种益生作用的研究。L. plantarum KH13-1具有调节抗生素所致菌群失调小鼠肠道菌群平衡的作用;摄入该菌后对免疫功能低下小鼠的免疫功能有增强作用,而对正常小鼠无显著影响;它的热致死菌体可促进小鼠脾细胞IL-4和IFN-γ的产生,但其活菌体对IL-4和IFN-γ的产生无影响。
In this study , the biological characteristic of 28 strains of Lactobacillus plantarum were analyzed, which isolated from traditional dairy products in pasture region of Inner Mongolia, such as tolerance to acid and bile salt, inhibition of their fermentation supernatant to the indicator . According to the screening experiment in vitro, L. plantarum KH13-1 was selected to a lot of studies, e.g checked the plasmid of L. plantarum KH13-1, its regulation function of immunity and intestinal flora of mice.
22 strains of L. plantarum showed resistance to acid and bile salt among of tested bacteria. All of them could grow in the medium contained 0.3 % sodium taurocholate, and show tolerance to acid pH3.0 in a certain degree. 9 strains of L. plantarum could resist acid well, survival rate can even reach 150.6%. The survival rate of other strains of bacteria was different from 23.4% to 39.8%; The 9 strains of L. plantarum had a wide antimicrobial activity to pathogenic bacteria, e.g Escherichia col, Staphylococcus aureus, Shigella flexneri, Micrococcus lysodeikticus, Bacillus cereus, et al. The antimicrobial activity was weak to nonpathogenic bacteria. The antimicrobial substance was mainly organic acid and. L. plantarum KH13-1 had no plasmid.
According to the screening experiment in vitro, L. plantarum KH13-1 was studied to regulation function of immunity and intestinal microflora of mice. L. plantarum KH13-1 could modulate the normal balance of intestinal microflora in mice with antibiotic-associated intestinal flora imbalance; And the results showed that the strain could enhance immune function of IDMM, but it had no effect on the specific immunity to normal mice, at the same time, its heat-killed bacteria could also induce IL-4 and IFN-γproduction by spleen cells from the mice, but live bacteria had no influence.
引文
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3 金世琳.乳酸菌的科学与技术[J]. 中国乳品工业,1998,26(2):14-15
4 Fuller,R.1989. Probiotics in man and animals.J.Appl. Bacteriol. 66:365-378;
5 Lee,Y.K.and salminen,S.l997.The coming of age of probiotics.Trends in Food Science and Technology 6:241-245
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7 Chanda,R.C.1999. Enhancing Market Value of Milk by AddingCultures. J.Dairy Sci.
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