黄芩苷调节肠道菌群抑制高脂饮食诱导的结直肠癌肝转移研究
Baicalin modulates gut microbiota and inhibits liver metastasis of colorectal cancer induced by high⁃fat diet
- 上海中医药杂志 2023年57卷第10期 页码：59-67
- 作者机构：
1.上海中医药大学中西医结合学院（上海 201203）
- 作者简介：
魏娇，女，硕士研究生，主要从事中西医结合防治肿瘤研究工作
赵玲，副研究员，硕士研究生导师；E-mail：lzhao@shutcm.edu.cn
- 基金信息：国家自然科学基金项目(82104955);上海市科委“科技创新行动计划”启明星项目(22QA1408700)
DOI：10.16305/j.1007-1334.2023.2304067
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魏娇,征宗梅,侯新新,等.黄芩苷调节肠道菌群抑制高脂饮食诱导的结直肠癌肝转移研究[J].上海中医药杂志,2023,57(10):59-67.
WEI Jiao,ZHENG Zongmei,HOU Xinxin,et al.Baicalin modulates gut microbiota and inhibits liver metastasis of colorectal cancer induced by high⁃fat diet[J].Shanghai Journal of Traditional Chinese Medicine,2023,57(10):59-67.
魏娇,征宗梅,侯新新,等.黄芩苷调节肠道菌群抑制高脂饮食诱导的结直肠癌肝转移研究[J].上海中医药杂志,2023,57(10):59-67. DOI： 10.16305/j.1007-1334.2023.2304067.
WEI Jiao,ZHENG Zongmei,HOU Xinxin,et al.Baicalin modulates gut microbiota and inhibits liver metastasis of colorectal cancer induced by high⁃fat diet[J].Shanghai Journal of Traditional Chinese Medicine,2023,57(10):59-67. DOI： 10.16305/j.1007-1334.2023.2304067.
摘要
目的,2,评估黄芩苷抑制结直肠癌肝转移的疗效并探讨其潜在的作用机制，为中医药防治结直肠癌转移提供科学依据。,方法,2,24只6周龄C57BL/6J雄性小鼠随机分为4组，每组6只，分别为正常饮食组、高脂饮食组、黄芩苷低剂量组（50 mg/kg，喂饲高脂饮食）和黄芩苷高剂量组（200 mg/kg，喂饲高脂饮食）。小鼠接受正常或高脂饮食喂养4周后构建结直肠癌肝转移模型，术后给药干预4周。实验期间记录各组小鼠的体质量。实验结束后采血并检测总胆固醇（TC）、三酰甘油（TG）、低密度脂蛋白胆固醇（LDL-C）和高密度脂蛋白胆固醇（HDL-C）水平；称量腹股沟白色脂肪组织（iWAT）、附睾白色脂肪组织（eWAT）及背部棕色脂肪组织（BAT）的质量；记录原位肿瘤质量、肝脏质量、肝转移灶数量，计算肝脏指数（肝质量/体质量）和肝转移率；采用苏木精-伊红（HE）染色观察肝脏组织形态学变化；利用16S rRNA测序分析小鼠肠道菌群变化；采用实时荧光定量逆转录聚合酶链式反应（RT-qPCR）检测小鼠肝转移灶中上皮间质转化（EMT）标志物波形蛋白（,Vimentin,）、N-钙黏蛋白（,N,-,cadherin,）、E-钙黏蛋白（,E,-,cadherin,）和闭合蛋白（,Occludin,）等相关基因mRNA的表达水平；采用皮尔森（Pearson）相关性分析方法检测肝转移灶中,EMT,基因表达水平与厚壁菌门细菌丰度的相关性。,结果,2,与正常饮食组比较，高脂饮食组小鼠体质量、血清TC和LDL-C水平、脂肪质量、原位肿瘤质量、肝质量、肝脏指数、肝转移灶数量及肝转移率明显增加，黄芩苷低、高剂量组上述指标均低于高脂饮食组，差异有统计学意义（,P,<,0.05）。高脂饮食组小鼠厚壁菌门丰度上升（,P,<,0.05），厚壁菌门/拟杆菌门比值增加（,P,<,0.05），经黄芩苷干预后，厚壁菌门/拟杆菌门比值下降（,P,<,0.05）。与正常饮食组比较，高脂饮食组小鼠肝转移灶,Vimentin,和,N,-,cadherin, mRNA表达水平明显升高（,P,<,0.05），,E,-,cadherin,和,Occludin, mRNA表达水平明显降低（,P,<,0.05）；与高脂饮食组比较，黄芩苷组小鼠肝转移灶,Vimentin,和,N,-,cadherin, mRNA表达水平明显降低（,P,<,0.05），,E,-,cadherin,和,Occludin, mRNA表达水平明显升高（,P,<,0.05）。厚壁菌门多种细菌相对丰度与,Vimentin,和,N,-,cadherin, mRNA表达水平呈正相关，但与,E,-,cadherin,和,Occludin, mRNA表达水平呈负相关。,结论,2,黄芩苷可有效抑制高脂饮食诱导的结直肠癌肝转移，且与肠道菌群结构的恢复、上皮间质转化的逆转相关。
Abstract
Objective,2,To investigate the efficiency and underlying mechanism of baicalin in the treatment of colorectal cancer liver metastasis（CRLM） so as to provide a scientific foundation for the prevention and treatment of CRLM with traditional Chinese medicine.,Methods,2,Twenty-four C57BL/6J male mice aged 6 weeks were randomly divided into 4 groups： the normal diet group， the high-fat diet group， the low-dose baicalin group （50 mg/kg， fed with a high-fat diet） and the high-dose baicalin group （200 mg/kg， fed with a high-fat diet） with 6 mice in each group. The liver metastasis of colorectal cancer model was established after the mice were fed with the normal or high-fat diet for 4 weeks. Subsequent to the surgery， the mice were orally gavaged with baicalin or PBS daily for 4 weeks. The body weights of the mice in each group were recorded during the experiment. At the end of the experiment， blood samples were collected for detecting the levels of total cholesterol （TC）， total triglyceride（TG）， low-density lipoprotein cholesterol（LDL-C） and high-density lipoprotein cholesterol （HDL-C）. The mass of inguinal white adipose tissue （iWAT）， epididymal white adipose tissue （eWAT） and dorsal brown adipose tissue （BAT） was weighed. Primary tumor weight， liver weight and the number of liver metastasis foci were recorded， and the liver index （the ratio of liver weight to body weight） and liver metastasis rate were calculated. HE staining was used to observe the morphological changes of the livers. The 16S rRNA gene sequence was used to analyze the structure and composition of the gut microbiota. The mRNA expression levels of epithelial mesenchymal transition （EMT） related genes including ,Vimentin,、,N,-,cadherin,、,E,-,cadherin,， and ,Occludin, were detected by RT-qPCR. The relationship between EMT related gene expression levels and Firmicutes microbiota abundance was determined using Pearson correlation analysis.,Results,2,The body weight， serum levels of TC and LDL-C， fat weight， primary tumor weight， liver weight， liver index， the number of liver metastases and liver metastases rate in the high-fat diet group were significantly higher than those of mice in the normal diet group. The data of the low-dose and high-dose baicalin groups were lower than those of the high-fat diet group. All of the above differences were statistically significant （,P,<,0.05）. Firmicutes abundance and the ratio of Firmicutes/Bacteroidetes （F/B） were increased （,P,<,0.05） in the high-fat diet group， and the F/B ratio decreased （,P,<,0.05） after baicalin treatment. Compared with the normal diet group， the mRNA expression levels of ,Vimentin, and ,N⁃cadherin, in liver metastases in the high-fat diet group were significantly increased （,P,<,0.05）， while the mRNA expression levels of ,E⁃cadherin, and ,Occludin, were significantly decreased （,P,<,0.05）； compared with the high-fat diet group， after intervention with baicalin， the mRNA expression levels of ,Vimentin ,and ,N⁃cadherin, were significantly decreased （,P,<,0.05）， while the mRNA expression levels of ,E‑cadherin, and ,Occludin ,were significantly increased （,P,<,0.05）. The relative abundances of several Firmicutes bacteria were positively correlated with the expression levels of ,Vimentin, and ,N⁃cadherin, mRNA， but negatively correlated with the expression levels of ,E⁃cadherin, and ,Occludin, mRNA levels.,Conclusion,2,Baicalin effectively inhibits high-fat diet-induced colorectal cancer liver metastasis， which is associated with the restoration of gut microbiota structure and the reversal of EMT.
关键词
结直肠癌肿瘤转移黄芩苷肠道菌群小鼠模型中药研究
Keywords
colorectal cancertumor metastasisbaicalingut microbiotamice modeltraditional Chinese herbal medicine research
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