基于16S rRNA测序技术研究口服益生菌对结肠癌皮下瘤小鼠肠道菌群结构的影响  

Effects of oral probiotics on gut microbiota structure in subcutaneous tumors in mice with colon cancer based on 16S rRNA sequencing technology

作　　者：张树玲[1] 王军威 左鲁玉 胡世亮 孙君志[1] 王纯[1] ZHANG Shuling;WANG Junwei;ZUO Luyu;HU Shiliang;SUN Junzhi;WANG Chun(Chengdu Sport University,Chengdu 610041,China)

机构地区：[1]成都体育学院,成都610041

出　　处：《中国实验动物学报》2023年第12期1564-1572,共9页Acta Laboratorium Animalis Scientia Sinica

基　　金：四川省科技计划项目资助(2022JDRC0137);运动医学四川省重点实验室2023年创新课题(2023-A014)。

摘　　要：目的基于16S rRNA测序技术探讨口服益生菌对结肠癌皮下荷瘤小鼠肠道菌群多样性、群落结构及组间差异的影响。方法24只6周龄BALB/c雄性小鼠,适应性喂养1周后,随机分为对照组(NC组,n=8),模型组(M组,n=8),益生菌+模型组(PM组,n=8)。PM组采用灌胃200μL益生菌混合溶液(长双歧杆菌和德氏乳杆菌保加利亚亚种混合冻干粉,2×10^(8)CFU),NC组、M组则给予200μL生理盐水,每周3次,连续7周。于10周龄末,M组和PM组左后肢侧背部皮下接种0.2 mL CT26.WT细胞悬液(浓度为1×10^(7)/mL),NC组相同部位接种0.2 mL生理盐水。观察小鼠一般状态、监测皮下肿瘤生长情况,利用16S rRNA测序法检测肠道菌群结构的变化。结果M组小鼠皮下瘤凸显,PM组小鼠皮下瘤体积与重量显著减少(P<0.05)。与NC组相比,M组小鼠肠道菌群Alpha多样性指数降低,组间Beta多样性存在显著性差异(P<0.01),补充益生菌对M组小鼠肠道菌群结构多样性具有一定的回调作用。与M组相比,PM组拟杆菌门(Bacteroidetes)、变形菌门(Proteobacteria)、Muribaculaceae、拟杆菌属(Bacteroides)等相对丰度升高,厚壁菌门(Firmicutes)、脱硫菌门(Desulfobacterota)、毛螺菌属NK4AB6(Lachnospiraceae_NK4A136_group)、另枝菌属(Alistipes)等相对丰度降低。LEfSe分析表明,PM组Muribaculaceae、拟杆菌属(Bacteroides)为丰度较高的差异物种(LDA值>4)。结论口服益生菌可能通过增加结肠癌皮下瘤小鼠肠道菌群中有益菌Muribaculaceae和拟杆菌属(Bacteroides)的相对丰度,进而改善肠道菌群失调状态。Objective To investigate the effects of oral probiotics on gut microbiota diversity,colony structure,and intergroup differences in mice with subcutaneous colon cancer tumors,based on 16S rRNA sequencing technology.Methods Twenty-four 6-week-old male BALB/c mice were divided randomly into normal control group(NC group,n=8),model group(M group,n=8),and probiotic+model group(PM group,n=8)after adaptive feeding for 1 week.Mice in the PM group were given 200μL probiotic mixed solution(Bifidobacterium longum and Lactobacillus delbrueckii subsp.bulgaricus mixed lyophilized powder,2×10^(8) colony-forming units)by gavage three times/week for 7 weeks,while the M group and PM group received 200μL normal saline.At 10 weeks old,0.2 mL CT26.WT cell suspension(1×10^(7)/mL)was inoculated subcutaneously into the left hind limbs of M group and PM group,while NC group were inoculated with 0.2 mL normal saline.The general state of mice was observed,the growth of subcutaneous tumor was monitored,and the changes of intestinal flora structure were detected by 16S rRNA sequencing.Results The subcutaneous tumors of the M group were prominent,and the subcutaneous tumor volume and weight of the PM group were significantly reduced(P<0.05).Compared with NC group,Alpha diversity index was lower in the M group,and a significant difference of Beta diversity inter groups(P<0.01).And supplementation of probiotics had a certain effect on gut microbiota diversity in the M group.Compared with M group,the relative abundance of Bacteroidetes,Proteobacteria,Muribaculaceae,Bacteroides were higher in the PM group,while the relative abundance of Firmicutes,Desulfobacterota,Lachnospiraceae_NK4A136_group,Alistipes were lower in the PM group.LEfSe analysis showed that Muribaculaceae and Bacteroides in the PM group were different species with high abundance(LDA values>4).Conclusions Oral probiotics may improve the gut microbiota by increasing the relative abundance of beneficial Muribaculaceae and Bacteroides in subcutaneous tumors in mice with colon canc

关 键 词：益生菌 结肠癌 皮下瘤 肠道菌群 

分 类 号：Q95-33[生物学—动物学]