具核梭杆菌灌胃对大鼠肠道菌群的影响

doi:10.6040/j.issn.1671-7554.0.2017.929

摘要/Abstract

摘要： 目的探究具核梭杆菌(Fusobacterium nucleatum)灌胃对健康大鼠肠道菌群的影响。方法将16只SD雄性大鼠随机分为2组(每组8只),处理组在大鼠第4、5、6、7、8周龄分别用具核梭杆菌灌胃1次,对照组在相同时间点以等量生理盐水灌胃。在大鼠第3、8、12周末收集粪便,提取DNA后通过Illumina Miseq平台进行16S rRNA基因高通量测序。应用蛋白印迹法检测大鼠粪便中针对具核梭杆菌的特异性IgA。结果第3周时,两组在菌群多样性、主要菌门水平方面的差异无统计学意义。第8周时,处理组菌群多样性明显低于对照组(P=0.009);处理组的厚壁菌门、变形菌门含量低于对照组,拟杆菌门含量高于对照组,差异均有统计学意义(P<0.05)。第12周时,处理组菌群多样性明显高于对照组(P=0.021),两组大鼠肠道菌群的主要菌门水平差异无统计学意义。同时,具核梭杆菌灌胃刺激机体产生了特异性IgA。结论具核梭杆菌通过未定植方式,对肠道菌群产生了影响,并可持续一定时间。具核梭杆菌刺激产生的特异性IgA可能是其不能在肠道定植的原因之一。

关键词: 具核梭杆菌, 肠道菌群, 高通量测序, 大鼠

Abstract: Objective To investigate the effects of Fusobacterium nucleatum(F. nucleatum)gavage on the intestinal microbiota of rats. Methods A total of 16 male SD rats were randomly divided into 2 groups: intervention group(n=8)and control group(n=8). The intervention group was treated with F. nucleatum when the rats were 4, 5, 6, 7 and 8 weeks old. The control group was treated with the same dosage of normal saline at the same time. Feces was collected at the end of week 3, 8 and 12 to extract DNA, and then Illumina Miseq platform was used for 16S rRNA gene sequencing. Specific IgA against F. nucleatum in stool was detected with Western blotting. Results There were no significant differences between the two groups in terms of fecal microbial diversity and relative abundance of main phyla in week 3. The microbial diversity of the intervention group was significantly lower than that of the control group in week 8(P=0.009). The relative abundances of Firmicutes and Proteobacteriodes in the intervention group were significantly less than those in the control group(P<0.05), while the relative abundance of Bacteriodes in the intervention group was significantly higher than that in the control group in week 8(P<0.05). In week 12, the intervention group showed higher diversity(P=0.021), and there was no significant difference in the relative abundance of main phyla. Moreover, the treatment of F. nucleatum stimulated the production of specific IgA. Conclusion The F. nucleatum can interfere with intestinal microbiota independent on colonization, and this effect can last for some time. The production of specific IgA induced by F. nucleatum may explain why this bacterium does not colonize in the intestine.

Key words: Fusobacterium nucleatum, Intestinal microbiota, High-throughput sequencing, Rats

中图分类号:

R574

宋立锦,顾湘,李理想,李铭,左秀丽,李延青. 具核梭杆菌灌胃对大鼠肠道菌群的影响[J]. 山东大学学报 (医学版), 2018, 56(7): 7-14.

SONG Lijin, GU Xiang, LI Lixiang, LI Ming, ZUO Xiuli, LI Yanqing. Effects of Fusobacterium nucleatum gavage on the intestinal microbiota of rats[J]. Journal of Shandong University (Health Sciences), 2018, 56(7): 7-14.

参考文献

[1] Han YW. Fusobacterium nucleatum: a commensal-turned pathogen[J]. Curr Opin Microbiol, 2015, 23:141-147.doi:10.1016/j.mib.2014.11.013.
[2] Strauss J, Kaplan GG, Beck PL, et al. Invasive potential of gut mucosa-derived Fusobacterium nucleatum positively correlates with IBD status of the host[J]. Inflamm Bowel Dis, 2011, 17(9):1971-1978.
[3] Han YW, Fardini Y, Chen C, et al. Term stillbirth caused by oral Fusobacterium nucleatum[J]. Obstet Gynecol, 2010, 115(2 Pt 2):442-445.
[4] Yang Y, Weng W, Peng J, et al. Fusobacterium nucleatum increases proliferation of colorectal cancer cells and tumor development in mice by activating Toll-like receptor 4 signaling to nuclear factor-kappaB, and up-regulating expression of microRNA-21[J]. Gastroenterology, 2017, 152(4):851-866.
[5] Swidsinski A, Dorffel Y, Loening-Baucke V, et al. Acute appendicitis is characterised by local invasion with Fusobacterium nucleatum/necrophorum[J]. Gut, 2011, 60(1):34-40.
[6] Gur C, Ibrahim Y, Isaacson B, et al. Binding of the Fap2 protein of Fusobacterium nucleatum to human inhibitory receptor TIGIT protects tumors from immune cell attack[J]. Immunity, 2015, 42(2):344-355.
[7] Rubinstein MR, Wang X, Liu W, et al. Fusobacterium nucleatum promotes colorectal carcinogenesis by modulating E-cadherin/beta-catenin signaling via its FadA adhesin[J]. Cell Host Microbe, 2013, 14(2):195-206.
[8] Nakagaki H, Sekine S, Terao Y, et al. Fusobacterium nucleatum envelope protein FomA is immunogenic and binds to the salivary statherin-derived peptide[J]. Infect Immun, 2010, 78(3):1185-1192.
[9] Abed J, Emgard JE, Zamir G, et al. Fap2 mediates Fusobacterium nucleatum colorectal adenocarcinoma enrichment by binding to tumor-expressed Gal-GalNAc[J]. Cell Host Microbe, 2016, 20(2):215-225.
[10] Yu T, Guo F, Yu Y, et al. Fusobacterium nucleatum promotes chemoresistance to colorectal cancer by modulating autophagy[J]. Cell, 2017, 170(3):548-563.e16.
[11] Stecher B, Hardt WD. Mechanisms controlling pathogen colonization of the gut[J]. Curr Opin Microbiol, 2011, 14(1):82-91.
[12] Levy M, Kolodziejczyk AA, Thaiss CA, et al. Dysbiosis and the immune system[J]. Nat Rev Immunol, 2017, 17(4):219-232.
[13] Simren M, Barbara G, Flint HJ, et al. Intestinal microbiota in functional bowel disorders: a Rome foundation report[J]. Gut, 2013, 62(1):159-176.
[14] Marchesi JR, Adams DH, Fava F, et al. The gut microbiota and host health: a new clinical frontier[J]. Gut, 2016, 65(2):330-339.
[15] Arumugam M, Raes J, Pelletier E, et al. Enterotypes of the human gut microbiome[J]. Nature, 2011, 473(7346):174-180.
[16] Tettamanti L, Gaudio RM, Cura F, et al. Prevalence of periodontal pathogens among italian patients with chronic periodontitis: a retrospective study on 2992 patients[J]. Oral Implantol(Rome), 2017, 10(1):28-36.
[17] Wang HF, Li LF, Guo SH, et al. Evaluation of antibody level against Fusobacterium nucleatum in the serological diagnosis of colorectal cancer[J]. Sci Rep, 2016, 6:33440. doi: 10.1038/srep33440.
[18] Edgar RC. Search and clustering orders of magnitude faster than BLAST[J]. Bioinformatics, 2010, 26(19):2460-2461.
[19] Lan Y, Wang Q, Cole JR, et al. Using the RDP classifier to predict taxonomic novelty and reduce the search space for finding novel organisms[J]. PLoS One, 2012, 7(3):e32491. doi: 10.1371/journal.pone.0032491
[20] Lundberg DS, Yourstone S, Mieczkowski P, et al. Practical innovations for high-throughput amplicon sequencing[J]. Nat Methods, 2013, 10(10):999-1002.
[21] Zhang C, Li S, Yang L, et al. Structural modulation of gut microbiota in life-long calorie-restricted mice[J]. Nat Commun, 2013, 4:2163. doi: 10.1038/ncomms3163.
[22] Kaplan CW, Lux R, Haake SK, et al. The Fusobacterium nucleatum outer membrane protein RadD is an arginine-inhibitable adhesin required for inter-species adherence and the structured architecture of multispecies biofilm[J]. Mol Microbiol, 2009, 71(1):35-47.
[23] Nwizu NN, Marshall JR, Moysich K, et al. Periodontal disease and incident cancer risk among postmenopausal women: results from the Womens Health Initiative Observational Cohort[J]. Cancer Epidemiol Biomarkers Prev, 2017, 26(8):1255-1265.
[24] Michaud DS, Liu Y, Meyer M, et al. Periodontal disease, tooth loss, and cancer risk in male health professionals: a prospective cohort study[J]. Lancet Oncol, 2008, 9(6):550-558.
[25] Barton MK. Evidence accumulates indicating periodontal disease as a risk factor for colorectal cancer or lymphoma[J]. CA Cancer J Clin, 2017, 67(3):173-174.
[26] Momen-Heravi F, Babic A, Tworoger SS, et al. Periodontal disease, tooth loss and colorectal cancer risk: results from the nurses’ health study[J]. Int J Cancer, 2017, 140(3):646-652.
[27] Bertrand KA, Shingala J, Evens A, et al. Periodontal disease and risk of non-Hodgkin lymphoma in the health professionals follow-up Study[J]. Int J Cancer, 2017, 140(5):1020-1026.
[28] 束蓉. 牙周病病因及治疗研究进展[J]. 上海交通大学学报(医学版), 2007,27(6):625-628. SHU Rong. Research advances in etiology and treatment of periodontal disease[J]. Journal of Shanghai Jiaotong University(Medical Science), 2007, 27(6):625-628.
[29] Saygun I, Nizam N, Keskiner I, et al. Salivary infectious agents and periodontal disease status[J]. J Periodontal Res, 2011, 46(2):235-239.
[30] Nakajima M, Arimatsu K, Kato T, et al. Oral administration of P. gingivalis induces dysbiosis of gut microbiota and impaired barrier function leading to dissemination of enterobacteria to the liver[J]. PLoS One, 2015, 10(7):e0134234. doi:10.1371/journal.pone.0134234.
[31] 赵力, 李理想, 陈飞雪, 等. 枯草芽孢杆菌灌胃对小鼠肠道菌群的影响[J]. 山东大学学报(医学版), 2017,55(10):28-35. ZHAO Li, LI Lixiang, CHEN Feixue, et al. Effect of Bacillus subtilis gavage on the intestinal microbiota of mice[J]. Journal of Shandong University(Health Sciences), 2017, 55(10):28-35.
[32] Dethlefsen L, McFall-Ngai M, Relman DA. An ecological and evolutionary perspective on human-microbe mutualism and disease[J]. Nature, 2007, 449(7164):811-818.
[33] Stecher B, Hardt WD. The role of microbiota in infectious disease[J]. Trends Microbiol, 2008, 16(3):107-114.
[34] Kabat AM, Srinivasan N, Maloy KJ. Modulation of immune development and function by intestinal microbiota[J]. Trends Immunol, 2014, 35(11):507-517.
[35] Macpherson AJ, Uhr T. Induction of protective IgA by intestinal dendritic cells carrying commensal bacteria[J]. Science, 2004, 303(5664):1662-1665.
[36] Norman JM, Handley SA, Baldridge MT, et al. Disease-specific alterations in the enteric virome in inflammatory bowel disease[J]. Cell, 2015, 160(3):447-460.
[37] Shah MS, DeSantis TZ, Weinmaier T, et al. Leveraging sequence-based faecal microbial community survey data to identify a composite biomarker for colorectal cancer[J]. Gut, 2018, 67(5):882-891.
[38] Monaco CL, Gootenberg DB, Zhao G, et al. Altered virome and bacterial microbiome in human immunodeficiency virus-associated acquired immunodeficiency syndrome[J]. Cell Host Microbe, 2016, 19(3):311-322.
[39] Kostic AD, Gevers D, Siljander H, et al. The dynamics of the human infant gut microbiome in development and in progression toward type 1 diabetes[J]. Cell Host Microbe, 2015, 17(2):260-273.
[40] Wright EK, Kamm MA, Teo SM, et al. Recent advances in characterizing the gastrointestinal microbiome in Crohn's disease: a systematic review[J]. Inflamm Bowel Dis, 2015, 21(6):1219-1228.
[41] 谢莉敏, 李丹, 程秀芳, 等. 益生菌的种类及其在人体营养保健中的应用研究[J]. 安徽农业科学, 2013(17):7694-7695. XIE Limin, LI Dan, CHENG Xiufang, et al. On species and application of probiotics in human nutrition and health care [J]. Journal of Anhui Agri, 2013, 41(17):7694-7695.

相关文章 15

[1]	吴逸南葛志明李方贺红姜虹张运. 自发性高血压大鼠肾脏血管紧张素转换酶2的表达[J]. 山东大学学报(医学版), 2209, 47(6): 5-.
[2]	祝林胡三元张光永丁祥就. 前列腺素E2对阻塞性黄疸大鼠小肠粘膜形态的保护作用[J]. 山东大学学报(医学版), 2209, 47(6): 12-.
[3]	孙涛张道来谢珊珊王玉卓冯玉新辛华. 酒精对原代培养的神经前体细胞间隙连接蛋白43表达的影响[J]. 山东大学学报(医学版), 2209, 47(6): 20-.
[4]	张道来孙涛谢珊珊王玉卓赵玲冯玉新辛华. 体外原代培养胎鼠大脑皮层神经元NMDAR1亚基表达的发育性变化[J]. 山东大学学报(医学版), 2209, 47(6): 28-32.
[5]	赵万霞,詹群璋,金婷,刘玉新,曲崇正,吴剑纯. 基于孟德尔随机化分析肠道菌群、血液代谢物和肥胖的因果关系[J]. 山东大学学报 (医学版), 2026, 64(4): 72-82.
[6]	徐晶晶,王新起,张洋,许旺旺,高进. P2X7受体抑制剂对青春期创伤后应激障碍大鼠行为及肠道菌群的影响[J]. 山东大学学报 (医学版), 2025, 63(4): 1-9.
[7]	王小磊,方骏,王安,朱武晖,史光军. 两样本孟德尔随机化分析肠道菌群与肝外胆管癌的因果关系[J]. 山东大学学报 (医学版), 2025, 63(4): 44-50.
[8]	杜凯豪,侯立朝,东小鸽,薛伟伟,何洁洁,罗兰明慧,蒋威,汪占金,王展. 东亚人肠道菌群与胰腺癌关系:基于孟德尔随机化方法的遗传学证据[J]. 山东大学学报 (医学版), 2025, 63(12): 44-52.
[9]	刘绍庭,张薇薇,冉茜,王建. 食品乳化剂与肠道健康[J]. 山东大学学报 (医学版), 2024, 62(8): 18-26.
[10]	赵子辉,王旭阳,张鹏,方宁宁,石端博,杨帆,杨晓玫,吴剑波. 经膈肌穿刺法快速准确评估大鼠肺动脉高压[J]. 山东大学学报 (医学版), 2024, 62(2): 20-28.
[11]	于丽,王弋嘉,杨勇,刘学焕,万雪花,包翠萍,苗蓓亮,李斯琪,李静,刘筠. 基于光学表面波技术探究具核梭杆菌介导的自噬与细胞折射率的关系[J]. 山东大学学报 (医学版), 2024, 62(10): 87-97.
[12]	陈泰宇,唐学贵,唐诗宇,张智彬,陈思敏,李敏,庞峻. 白头翁汤对短肠综合征小鼠肠道功能的影响[J]. 山东大学学报 (医学版), 2023, 61(9): 29-37.
[13]	祁少俊,唐延金,张正铎,吴虹,张佳程,秦川,刘锐,高希宝. 补充多种微量元素对高糖饮食大鼠的保护作用[J]. 山东大学学报 (医学版), 2023, 61(7): 19-26.
[14]	管伟,白云峰,杜胜杰,王月兰,李希明. 基于RNA高通量测序研究右美托咪定对人神经母细胞瘤细胞基因表达的影响[J]. 山东大学学报 (医学版), 2023, 61(7): 27-33.
[15]	华月帆,何珂瑶,张家豪,钱梦凡,刘怡文,孔金玉,杨海军,周福有. 具核梭杆菌诱导缺氧诱导因子及血管生成因子高表达对食管鳞癌患者生存预后的影响[J]. 山东大学学报 (医学版), 2023, 61(11): 59-67.

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed