MNNG胃癌大鼠肠道菌群与转录组学特征分析  

Characterization of intestinal flora and transcriptomics in rats with gastric cancer

作　　者：张晨曦 李继娟 张飞程 高田宇 梁新月 潘丽佳 ZHANG Chenxi;LI Jijuan;ZHANG Feicheng;GAO Tianyu;LIANG Xinyue;PAN Lijia(College of Acupuncture and Massage,Hebei University of Traditional Chinese Medicine,Shijiazhuang 050200,China)

机构地区：[1]河北中医药大学针灸推拿学院,石家庄050200

出　　处：《中国实验动物学报》2025年第1期70-81,共12页Acta Laboratorium Animalis Scientia Sinica

基　　金：河北省自然科学基金(H2023423029);河北省中医药管理局科研计划项目(2022361,2021095);河北中医药大学2022年博士科研基金项目(BSZ2022006)。

摘　　要：目的分析N-甲基-N-硝基-N-亚硝基胍(N-methyl-N’-nitro-N-nitrosoguanidine,MNNG)胃癌大鼠与正常大鼠肠道菌群差异、转录组学差异,以探究MNNG胃癌大鼠肠道菌群与转录组学特征,并分析二者的相关性,为以MNNG胃癌大鼠为模型的相关研究提供参考。方法12只Wistar大鼠随机分为正常(Normal,NM)组、胃癌(Gastric cancer,GC)组,GC组给予浓度20 mg/mL的MNNG灌胃,剂量为100 g/mL,每天1次,NM组给予相同剂量的生理盐水灌胃,连续干预16周后取材检测。取大鼠胃组织,苏木素-伊红(HE)染色观察两组大鼠胃黏膜形态变化,转录组测序检测差异基因的表达水平;无菌Ep管收集盲肠内容物用于16S rRNA测序。结果(1)肉眼观察及HE结果:肉眼观察,NM组大鼠胃组织黏膜体积正常,表面有光泽,胃壁有弹性,黏膜皱襞走向有规律,无增生无出血点,GC组大鼠胃组织胃黏膜体积缩小,胃壁变薄,弹性变差,皱襞走向错乱无规律,出现隆起且伴有黄黑色角质增生;HE染色,NM组胃黏膜鳞状上皮层、粘膜下层、粘膜肌层层次清晰,无增生和角化,GC组胃粘膜层次和细胞极性紊乱,细胞形态不一,鳞状上皮层被破坏,鳞状上皮细胞增生和角化,并向下增殖侵入肌层,造模成功;(2)肠道菌群测序结果显示,MNNG胃癌大鼠阿克曼氏菌和乳杆菌的丰度显著减少,瘤胃球菌科、普雷沃氏菌、布劳特氏菌的丰度显著增加;(3)转录组学测序:京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析得到的3条关键通路分别是阿米巴病、系统性红斑狼疮和酯酰肌醇3激酶-丝氨酸/苏氨酸蛋白激酶(phosphatidylinositol 3 kinase-RAC-αserine/threonine-protein kinase,PI3K-Akt)信号通路,富集到这3条通路的5个差异基因分别是MCPT8I2、IGH-6、IGHG1、ACTN2、VEGF-D;(4)肠道菌群与转录组学联合分析结果显示,瘤胃球菌科_UCG-005、普雷沃氏菌_UCG-003和布劳特氏菌可能与阿米巴病、系统�Objective To analyze differences in the intestinal microbiota and transcriptomics between N-methyl-N’-nitro-N-nitrosoguanidine(MNNG)gastric cancer rats and normal rats and to analyze the correlation between the two,so as to provide a reference for related studies using MNNG gastric cancer rats as a model.Methods A total of 12 Wistar rats were randomly divided into normal(NM)and gastric cancer(GC)groups.The GC group was given a concentration of 20 mg/mL of MNNG by gavage with a dose of 100 g/mL once a day,and the NM group was given the same amount of normal saline by gavage.Samples were collected for testing after 16 weeks of continuous intervention.The gastric tissues were collected and stained by HE staining to observe morphological changes in the gastric mucosa of the two groups,and the expression levels of differential genes were detected by transcriptome sequencing.The cecal contents were collected for 16S rRNA sequencing.Results(1)Visual observation and HE results showed that the volume of gastric mucosa in the NM group was normal,the surface was glossy,the gastric wall was elastic,the direction of the mucosal folds was regular,there were no hyperplasia or hemorrhagic spots.In the GC group,the volume of gastric mucosa was reduced,the gastric wall was thinned,elasticity was poor,the direction of the folds was disordered and irregular,and there was a bulge accompanied by yellow-black keratotic hyperplasia.In the NM group,the squamous epithelial layer,submucosa,and muscular layer of the gastric mucosa were clear,with no hyperplasia and keratinization.In the GC group,the gastric mucosa had disorganized layers and cell polarity,with different cell morphologies;the squamous epithelial layer was destroyed,and squamous epithelial cells were hyperplasic,keratinized,and had invaded the muscular layer by proliferation.The modeling was considered successful.(2)The results of intestinal microbiota sequencing showed that the abundance of Akkermansia and Lactobacillus in MNNG gastric cancer rats decreased significantly,a

关 键 词：胃癌 胃癌模型 肠道菌群 转录组学 

分 类 号：Q95-33[生物学—动物学]