一株牦牛源屎肠球菌对小鼠肠道微生物、形态和免疫功能的影响
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摘要
为研究牦牛肠黏膜源屎肠球菌G2对健康小鼠肠道微生物、形态和免疫功能的影响,试验设置对照组(CON)、热灭活菌组(EHK)、活菌组(ENT)3个处理组,每组8只C57BL/6小鼠,共计24只。CON组、EHK组、ENT组于正式试验期第1、2、4、6天按0.1 mL/只分别灌胃M17培养基、热灭活菌液和活菌菌液,灌胃前无菌收集小鼠粪便进行乳酸菌总菌计数,第8天处死小鼠取样。结果表明:ENT组乳酸菌总菌数量在第3、5天显著大于CON组与EHK组(P <0.05);各组小鼠结肠病理学评分无显著差异,但ENT组隐窝深度显著高于CON组与EHK组(P <0.05);各组间肠黏膜免疫因子Reg3β、Reg3γ和MUC2基因mRNA表达量差异不显著,TNF-α表达量ENT组显著低于CON组(P <0.05)。综上可知,牦牛源屎肠球菌G2可能为一种肠道过路菌,能在短时间内提高肠道乳酸菌的总数,一定程度上具有改善肠道形态和促进肠道健康的作用。
The current study was conducted to evaluate effects of Enterococcus faecium G2,a probiotic strain isolated from yak intestinal mucosa,on intestinal microbial population,morphology and immune functions in healthy mice. A total of 24 C57 BL/6 mice(SPF level) randomly allocated into 3 groups:control group(CON),heat-killed Enterococcus faecium G2 group(EHK) and Enterococcus faecium G2 group(ENT). On day 1,2,4 and 6 of the trial,mice from CON,EHK and ENT groups were orally gavaged with0.1 mL M17 medium,heat-killed bacterial suspension and living bacterial solution,respectively. Mouse feces were collected before gavage to enumerate the total Lactic acid bacteria(LAB) counts. The colonic contents and tissues were collected after execution of the mice on day 8. The LAB counts of the ENT group was significantly higher than that of CON and EHK groups on day 3 and 5(P < 0.05). Intestinal pathological scores showed no significant difference. However crypt depth of the ENT group mice was significantly greater than the other two groups(P < 0.05).The relative mRNA expression of Reg3β,Reg3γ and MUC2 genes showed no significant difference among groups.Enterococcus faecium G2 group treated mice showed dramatically lower mRNA expression level of Tumor necrosis factor alpha(TNF-α) when compared to the control group(P < 0.05).The results indicated that Enterococcus faecium G2 could act as a potential passing bacteria,which induced a short-term increase of the fecal LAB counts,and could improve the intestinal morphology and promote health to a certain extent.
The current study was conducted to evaluate effects of Enterococcus faecium G2,a probiotic strain isolated from yak intestinal mucosa,on intestinal microbial population,morphology and immune functions in healthy mice. A total of 24 C57 BL/6 mice(SPF level) randomly allocated into 3 groups:control group(CON),heat-killed Enterococcus faecium G2 group(EHK) and Enterococcus faecium G2 group(ENT). On day 1,2,4 and 6 of the trial,mice from CON,EHK and ENT groups were orally gavaged with0.1 mL M17 medium,heat-killed bacterial suspension and living bacterial solution,respectively. Mouse feces were collected before gavage to enumerate the total Lactic acid bacteria(LAB) counts. The colonic contents and tissues were collected after execution of the mice on day 8. The LAB counts of the ENT group was significantly higher than that of CON and EHK groups on day 3 and 5(P < 0.05). Intestinal pathological scores showed no significant difference. However crypt depth of the ENT group mice was significantly greater than the other two groups(P < 0.05).The relative mRNA expression of Reg3β,Reg3γ and MUC2 genes showed no significant difference among groups.Enterococcus faecium G2 group treated mice showed dramatically lower mRNA expression level of Tumor necrosis factor alpha(TNF-α) when compared to the control group(P < 0.05).The results indicated that Enterococcus faecium G2 could act as a potential passing bacteria,which induced a short-term increase of the fecal LAB counts,and could improve the intestinal morphology and promote health to a certain extent.
引文
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[14]王利红.鸡源乳酸菌的分离筛选及其对肉鸡生产性能的影响[D].杨凌:西北农林科技大学,2014.
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[18]闫蕾,赵彩彦.肝脏-一个新的免疫器官[J].国际内科学杂志,2009,36(4):206~210.
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[20]张艳.高产细菌素乳酸菌的分离鉴定及其相关基因的克隆[D].成都:西南民族大学,2013.
[21]朱惠.牦牛源海氏肠球菌Swun3株对小鼠生长性能和肠黏膜免疫的影响[D].成都:西南民族大学,2016.
[22]邹立军,熊霞,王小城,等.肠道隐窝-绒毛轴上皮细胞更新及调控机制研究进展[J].中国科学:生命科学,2017,47(2):190~200.
[23]Ju T,Shoblak Y,Gao Y,et al.Initial Gut Microbial Composition as a Key Factor Driving Host Response to Antibiotic Treatment,as Exemplified by the Presence or Absence of Commensal Escherichia coli[J].Appl Environ Microbiol,2017,83(17):AEM.01107~17.
[24]Petersson,K.J.Domig,K.Schedle,et al.Compar ison of three methods to enumerate gut microbiota of weanling piglets fed insoluble dietary fibre differing in lignin content[J].Journal of Agricultural Science 2010,148:225~232.
[25]Rinttil a..T,Kassinen A,Malinen E,et al.Development of an extensive set of 16S rDNA-targeted primers for quantification of pathogenic and indigenous bacteria in faecal samples by real-time PCR[J].Journal of Applied Microbiology,2004,97(6):1166~1177.
[26]Wlodarska M,Willing B,Keeney K M,et al.Antibiotic treatment alters the colonic mucus layer and predisposes the host to exacerbated Citrobacter rodentium-induced colitis[J].Infection&Immunity,2011,79(4):1536~1545.
[27]Yan F,Cao H,Cover T L,et al.Soluble Proteins Produced by Probiotic Bacteria Regulate Intestinal Epithelial Cell Survival and Growth[J].Gastroenterology,2007,132(2):562~575.
[28]Zhao H W,Yue Y H,Han H,et al.Effect of toll-like receptor3 agonist poly I:C on intestinal mucosa and epithelial barrier function in mouse models of acute colitis[J].世界胃肠病学杂志(英文版),2017,23(6):999~1009.
[2]陈振,谢全喜,亓秀晔,等.复合益生菌替代抗生素对断奶仔猪生长性能、胃肠道pH和免疫器官指数的影响[J].中国畜牧杂志,2017,53(4):112~115.
[3]丁爽,郭春华,张正帆,等.产乳酸菌素屎肠球菌B13对断奶仔猪生长性能、养分消化率、血清免疫指标及粪便微生物菌群的影响[J].畜牧兽医学报,2017,48(10):1902~1911.
[4]董晓丽.益生菌的筛选鉴定及其对断奶仔猪、犊牛生长和消化道微生物的影响[D].北京:中国农业科学院,2013.
[5]方伟.屎肠球菌SF68对断奶仔猪生产性能、肠道形态和微生物区系的影响[D].扬州:扬州大学,2017.
[6]高彦华.猪抗菌肽PR-39的表达特性和调控及免疫调节机制研究[D].杭州:浙江大学,2014.
[7]侯成立,季海峰,周雨霞,等.植物乳杆菌对断奶仔猪生产性能和生化指标的影响[J].饲料研究,2011,12:14~16.
[8]胡远亮.利用分子生物技术研究益生菌对断奶仔猪生长及粪便菌群的影响[D].武汉:华中农业大学,2014.
[9]黄怡.屎肠球菌EF1对仔猪小肠黏膜屏障功能的影响[D].杭州:浙江大学,2012.
[10]李雨宸.RegIIIγ在乳酸菌拮抗结肠炎黏膜免疫中的作用研究[D].长春:吉林农业大学,2015.
[11]刘丹,卢放根.益生菌对小鼠急性溃疡性结肠炎模型疗效观察及机制研究[J].中国医学工程,2008,16(3):180~182.
[12]谯仕彦,候成立,曾祥芳.乳酸菌对猪肠道屏障功能的调节作用及其机制[J].动物营养学报2014,26(10):3052~3063.
[13]唐志刚.益生菌对饲喂不同营养水平日粮肉鸡消化和免疫功能的影响[D].南京:南京农业大学,2010.
[14]王利红.鸡源乳酸菌的分离筛选及其对肉鸡生产性能的影响[D].杨凌:西北农林科技大学,2014.
[15]王晓成,孙亚楠,任发政,等.益生菌对健康小鼠肠道形态发育的影响[J].中国奶牛,2017,6:49~54.
[16]王永,杨维仁,张桂国.饲粮中添加屎肠球菌对断奶仔猪生长性能、肠道菌群和免疫功能的影响[J].动物营养学报2013,25(5):1069~1076.
[17]徐基利.不同乳酸菌及其添加量对肉仔鸡生长性能、盲肠菌群和免疫功能的影响[D].哈尔滨:东北农业大学,2011.
[18]闫蕾,赵彩彦.肝脏-一个新的免疫器官[J].国际内科学杂志,2009,36(4):206~210.
[19]杨利娜,边高瑞,朱伟云.单胃动物肠道微生物菌群与肠道免疫功能的相互作用[J].微生物学报,2014,54(5):480~486.
[20]张艳.高产细菌素乳酸菌的分离鉴定及其相关基因的克隆[D].成都:西南民族大学,2013.
[21]朱惠.牦牛源海氏肠球菌Swun3株对小鼠生长性能和肠黏膜免疫的影响[D].成都:西南民族大学,2016.
[22]邹立军,熊霞,王小城,等.肠道隐窝-绒毛轴上皮细胞更新及调控机制研究进展[J].中国科学:生命科学,2017,47(2):190~200.
[23]Ju T,Shoblak Y,Gao Y,et al.Initial Gut Microbial Composition as a Key Factor Driving Host Response to Antibiotic Treatment,as Exemplified by the Presence or Absence of Commensal Escherichia coli[J].Appl Environ Microbiol,2017,83(17):AEM.01107~17.
[24]Petersson,K.J.Domig,K.Schedle,et al.Compar ison of three methods to enumerate gut microbiota of weanling piglets fed insoluble dietary fibre differing in lignin content[J].Journal of Agricultural Science 2010,148:225~232.
[25]Rinttil a..T,Kassinen A,Malinen E,et al.Development of an extensive set of 16S rDNA-targeted primers for quantification of pathogenic and indigenous bacteria in faecal samples by real-time PCR[J].Journal of Applied Microbiology,2004,97(6):1166~1177.
[26]Wlodarska M,Willing B,Keeney K M,et al.Antibiotic treatment alters the colonic mucus layer and predisposes the host to exacerbated Citrobacter rodentium-induced colitis[J].Infection&Immunity,2011,79(4):1536~1545.
[27]Yan F,Cao H,Cover T L,et al.Soluble Proteins Produced by Probiotic Bacteria Regulate Intestinal Epithelial Cell Survival and Growth[J].Gastroenterology,2007,132(2):562~575.
[28]Zhao H W,Yue Y H,Han H,et al.Effect of toll-like receptor3 agonist poly I:C on intestinal mucosa and epithelial barrier function in mouse models of acute colitis[J].世界胃肠病学杂志(英文版),2017,23(6):999~1009.