菌群移植对氟尿嘧啶抗鼠源结肠肿瘤的协同作用  

Synergistic effect of fecal microbiota transplantation on fluorouracil against colon tumor in mice

作　　者：杨秋雨 孙宇航[2] 檀梦天 白净[2] 李磊[2] 杨勇 吴芳[2] 李贺 付泽娴 YANG Qiuyu;SUN Yuhang;TAN Mengtian;BAI Jing;LI Lei;YANG Yong;WU Fang;LI He;FU Zexian(Hebei University of Engineering,Hebei Handan 056000,China;Affiliated Hospital of Hebei University of Engineering,Hebei Handan 056038,China)

机构地区：[1]河北工程大学,河北邯郸056000 [2]河北工程大学附属医院,河北邯郸056038

出　　处：《现代肿瘤医学》2024年第17期3213-3219,共7页Journal of Modern Oncology

基　　金：河北省教育厅科学研究项目(编号:ZD2022039)。

摘　　要：目的:观察菌群移植对氟尿嘧啶抑制结肠癌MC38荷瘤小鼠移植瘤生长的协同作用效果。方法:选取雌性C57BL/6N小鼠21只作为研究对象,分为空白组、对照组、实验组。培养MC38小鼠结肠癌细胞接种于小鼠右前肢背部皮下构建结肠癌荷瘤模型。空白组无任何处理,取其粪便制作干预菌液,实验组和对照组自分组后每两天1次给予0.1 mL菌液/生理盐水灌胃至实验结束。成瘤后第6天,按照65 mg/kg剂量连续5天腹腔注射氟尿嘧啶,3天后处死实验动物,计算肿瘤体积和抑瘤率。采用HE染色观察结直肠组织情况。采用酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)法检测白细胞介素-6(interleukin-6,IL-6)和肿瘤坏死因子Alpha(tumor necrosis factor alpha,TNF-α)血清水平,通过16S测序进行肠道菌群检测和分析。结果:与空白组相比,实验组和对照组模型小鼠体重均明显降低(P<0.05),但对照组和实验组体重变化未见明显差异(P>0.05)。对照组荷瘤平均体积大于实验组荷瘤平均体积(P<0.05)。与对照组相比,实验组结直肠组织病理镜下可见更多的隐窝细胞,绒毛更完整。实验组IL-6血清水平高于对照组和空白组(P<0.05);实验组和对照组TNF-α血清水平与空白组相比升高明显(P<0.05),但实验组和对照组差异无统计学意义(P>0.05)。16S测序结果提示自体菌群移植干预后,实验组肠道菌群更接近空白组,对照组菌群丰富度最低,但三组未见明显区别(P>0.05)。进一步分析三组菌群差异,发现在属(Genus)水平上,与空白组相比,实验组的Mar-Odoribacter和Tan-Parabacteroide富集增多,对照组Ent-Escherichia-Shigella富集增多。结论:菌群移植后上调Mar-Odoribacter和Tan-Parabacteroide菌,下调Ent-Escherichia-Shigella菌,可保护肠道黏膜,改善机体代谢,激活机体免疫系统,促进了IL-6和TNF-α细胞因子表达,提高抗肿瘤免疫能力,对氟尿嘧啶抑制MC38荷瘤小鼠移植瘤增殖起到了�Objective:To observe the synergistic effect of fecal microbiota transplantation on the inhibition of graft tumor growth by fluorouracil in MC38 mouse colon cancer cells loaded mice.Methods:21 female C57BL/6N mice were selected as study subjects and divided into blank group,control group and experimental group.MC38 mouse colon cancer cells were used to inoculate the mice subcutaneously on the back of the right forelimb to construct a colon cancer hormonal tumor model.The blank group was treated without any treatment,and its feces were taken to make the intervention bacterial solution,and the experimental and control groups were given 0.1 mL bacterial solution/saline gavage once every two days since the grouping until the end of the experiment.On the 6th day after tumor formation,fluorouracil was injected intraperitoneally for 5 consecutive days at a dose of 65 mg/kg,and the experimental animals were executed after 3 days to calculate the tumor volume and tumor suppression rate.The colorectal tissues were observed by HE staining.Serum levels of interleukin-6( IL-6) and tumor necrosis factor alpha( TNF-α) were detected by enzyme-linked immunosorbent assay( ELISA),and intestinal flora was detected and analyzed by 16S sequencing.Re sults:Compared with the blank group,the body weight of the model mice in both the experimental and control groups was significantly reduced( P < 0.05),but no significant difference in body weight change was observed between the control and experimental groups( P > 0.05).The average volume of the control group's loaded tumors was larger than the average volume of the experimental group 's loaded tumors( P < 0.05).Compared with the control group,more crypt cells with more intact villi were seen microscopically in the histopathologic histological sections of colorectal tissue in the experimental group.The serum level of IL-6 in the experimental group was higher than that in the control group and the blank group( P < 0.05).The serum level of TNF-α in the experimental group and the control gro

关 键 词：粪群移植 肿瘤免疫 荷瘤小鼠 皮下移植瘤模型 

分 类 号：R735.3[医药卫生—肿瘤]