基于微生物组学探讨肠复方抑制裸鼠肠癌原位移植瘤的作用机制  

Exploring the Mechanism on Inhibition of Orthotopic Transplantation of Intestinal Cancer in Nude Mice by Intestinal Compound Based on Microbiomics

作　　者：梁慧[1] 王莹雪 马璐文 谭骏岚 刘赳 LIANG Hui;WANG Yingxue;MA Luwen;TAN Junlan;LIU Jiu(Hunan Cancer Hospital,Changsha 410013 Hunan,China;Hunan University of Chinese Medicine,Changsha 410208 Hunan,China)

机构地区：[1]湖南省肿瘤医院,湖南长沙410013 [2]湖南中医药大学,湖南长沙410208

出　　处：《中药新药与临床药理》2025年第1期103-113,共11页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：湖南省中医药管理局重点项目(2021035);湖南省“十四五”第二批中医药学科带头人项目。

摘　　要：目的基于微生物组学探讨肠复方抑制裸鼠肠癌原位移植瘤的作用机制。方法复制荧光标记HCT-116肠癌细胞原位移植瘤裸鼠模型。术后第10天,采用小动物活体成像系统检测小鼠腹腔荧光信号。将模型复制成功的小鼠随机分为模型组、肠复方组(36g·kg^(-1))、抗生素组、抗生素联合肠复方组(简称联合组),另取健康裸鼠作为空白对照组,每组6只。灌胃给药(20mL·kg^(-1)),每日1次,连续干预4周。实验期间每2d称小鼠体质量并记录;采用小动物活体成像系统评估小鼠体内肿瘤生长情况;HE染色法观察肠癌组织病理变化;免疫组化法检测肠癌组织Ki67表达水平;采集小鼠粪便,进行肠道菌群16S rDNA扩增子测序及生物信息学分析。结果(1)与空白对照组比较,模型组小鼠的体质量显著降低(P<0.001)。与模型组比较,肠复方组小鼠体质量显著升高(P<0.01),肠癌细胞总荧光值明显降低(P<0.05),肠癌组织中Ki67表达显著下调(P<0.001);抗生素组小鼠体质量明显降低(P<0.05),肠癌细胞总荧光值有升高趋势,但差异无统计学意义(P>0.05),肠癌组织中Ki67表达显著上调(P<0.01)。与抗生素组比较,联合组小鼠体质量显著升高(P<0.001),肠癌细胞总荧光值明显降低(P<0.05),肠癌组织中Ki67表达显著下调(P<0.001)。(2)与空白对照组比较,模型组的chao1指数明显下降(P<0.05),变形菌门占比明显增加(P<0.05),屎肠球菌、克雷伯杆菌占比明显增加(P<0.05),肠道菌群的双组分系统丰度明显上升(P<0.05)。与模型组比较,肠复方组的chao1、simpson指数均无明显变化(P>0.05),疣微菌门占比增加(P>0.05),Akk菌占比增加(P>0.05),LEfSe分析显示疣微菌门、Akk菌为优势菌种,肠道菌群的双组分系统及肽酶丰度明显下降(P<0.05);抗生素组的simpson指数明显下降(P<0.05),chao1指数无明显变化(P>0.05),变形菌门占比显著增加(P<0.01),克雷伯杆菌占比显著增加(P<0.01),拟杆菌占比明显Objective To investigate the mechanism of the inhibitory effect of intestinal compound on orthotopic transplantation of intestinal cancer in nude mice through the analysis of microbiomics.Methods The orthotopic transplantation nude mouse model of fluorescently labeled HCT-116 intestinal cancer cells was established.On the 10 th day of post-operation,small animal imaging system was used to detect fluorescence signals in the abdominal cavity of mice.The successfully modeled mice were randomly divided into model group,intestinal compound group(36 g·kg^(-1)),antibiotic group,antibiotic-combined intestinal compound group,and blank control group,with 6 mice in each group.The corresponding drug was administered at 20 mL·kg^(-1)/day orally for 4 consecutive weeks.The body weight of mice in each group were observed at every two day intervals.Tumor growth in mice was evaluated by using small animal imaging system.HE staining was used for detection of histopathological changes in various groups of intestinal cancer,and immunohistochemistry was applied for detection of Ki67 expression.Feces from mice in each group were collected for high throughput 16S rDNA amplicon sequencing and bioinformatic analysis.Results(1)Compared with the blank group,the body weight of mice in the model group was significantly decreased(P<0.001).Compared with the model group,the body weight of mice in the intestinal compound group was elevated(P<0.01),the total fluorescence value and Ki67 expression of intestinal cancer tissue in the intestinal compound group obviously decreased(P<0.05,P<0.001).The body weight of the antibiotic group was reduced(P<0.05),and there was a trend of increase in the total fluorescence value of intestinal cancer cells,but the difference was not statistically significant(P<0.05).Ki67 expression of intestinal cancer tissue significantly up-regulated(P<0.01).Compared with the antibiotic group,the body weight of the mice in the combination group was elevated(P<0.001),the total fluorescence value of intestinal cancer cells si

关 键 词：肠癌 肠复方 抗生素 肠道菌群 微生物组学 Akk菌 HCT-116肠癌细胞 裸鼠 

分 类 号：R285.5[医药卫生—中药学]