高脂状态下肠道菌群结构改变与大肠癌发生关系的实验研究  

Experimental study about the relationship between the change of the structure of intestinal flora and the occurrence of colorectal cancer by the intervention of high-fat diet

作　　者：朱庆超[1] 汪昱[1] ZHU Qing-chao;WANGYu(Department of General Surgery,The Sixth People＇s Hospital Amliated to Shanhai Jiao-tong University,Shanghai 200233,China)

机构地区：[1]上海交通大学附属第六人民医院普外科,上海200233

出　　处：《中国现代普通外科进展》2018年第10期757-763,共7页Chinese Journal of Current Advances in General Surgery

基　　金：国家自然科学基金资助项目(No:81230057)

摘　　要：目的:构建大肠癌的动物模型,观察高脂饮食干预对结肠肿瘤发生情况的影响,对高脂饮食干预下肠道菌群结构的改变及其与大肠癌发生的关系进行初步探讨。方法:4周龄的雄性Wistar大鼠140只随机分为4组:普通饮食+EDTA对照的非诱导肿瘤组(SDC)、普通饮食+DMH的诱导肿瘤组(SDT)、高脂饮食+EDTA对照的非诱导肿瘤组(HFDC)、高脂饮食+DMH的诱导肿瘤组(HFDT)。根据组别的不同分别给予13.5%脂肪含量的普通饮食和45%脂肪含量的高脂饮食,诱导肿瘤组大鼠每周腹腔注射1,2-二甲基肼(DMH),剂量为40 mg/kg,连续注射10周,对照组大鼠每周腹腔注射同等剂量的EDTA+生理盐水。实验结束后将收集的肿瘤组织标本利用QIAamp DNA Mini试剂盒进行基因组DNA抽提,后续基因组DNA鉴定、PCR扩增、荧光定量、16S rRNA V3可变区测序和生物信息学分析;比较高脂饮食与普通饮食干预下组织中菌群结构的差异及其与肿瘤发生情况的关系。结果:饮食干预后的焦磷酸测序结果显示高脂能够导致肠道菌群结构失衡,增加大肠癌的发病风险;经LEfSe分析发现梭状杆菌属Fusobacterium可被归为组间最主要的差异性菌属。结论:高脂饮食能够改变肠道菌群结构,增加多种潜在性致病菌丰度,特别是梭状杆菌属,结合高脂状态下肿瘤的发生情况,提示该菌属在大肠癌的发生发展中起重要的作用。Objective： To observe the effect of high-fat diet on the incidence of colorectalcancer, and to investigate the change of intestinal flora and the relationship between them. Meth-ods： The grouping method and the diet intervention was 4 week old male Wistar rats were slectedand randomly divided into four groups： the normal diet ＋ EDTA control of non induced tumor group（SDC）, normal diet ＋DMH induced tumor group（SDT）, high fat diet ＋ EDTA control non induced tumorgroup （HFDC）, high fat diet ＋DMH induced tumor group （with video associated. According to thegroups rats were given 13.5%fat content of normal diet group and 45% fat content of high fat diet,the rats of tumor group undergone intraperitoneal injection of 1,2-dimethyl hydrazine （DMH） induceddose 40mg/kg every week for 10 weeks, the control group rats weekly by intraperitoneal injection ofthe same dose could saline. After the tissue samples were collected, genomic DNA was extractedusing the QIAamp DNA Mini Kit according to the manufacturer’s instructions. Afterwards, the determinationof genomic DNA, PCR amplification, fluorescence quantitation, pyrosequencing of 16SrRNA V3 region and the bioinformatic analysis were performed as described above in order to determinethe key bacterial phylotypes that might play important roles in the development of CRC; Re-sults： There was marked difference in the microbial composition of cancerous tissues fromdifferentdiet-treated rats. And we indicated that Fusobacterium could be classified as the key phylotypewhich contributing to the separation of microbiota in CRC rats and healthy rats. Conclusion： Combinedwith the occurrence of tumors in the state under high-fat, our results suggested that thisspecies might play an important role in the development of colorectal cancer.

关 键 词：结肠肿瘤 动物模型 高脂饮食 菌群失衡揖 

分 类 号：R735.35[医药卫生—肿瘤]