藏族药二十五味松石丸调控肠道菌群改善非酒精性脂肪性肝炎的作用机制研究  

Mechanism of Tibetan medicine Ershiwuwei Songshi Pills in regulating intestinal flora and improving non-alcoholic steatohepatitis

作　　者：简程芳 张博宇 王存萍 沙玉茹 李阿溶 龚普阳 顾健 谭睿[2] JIAN Cheng-fang;ZHANG Bo-yu;WANG Cun-ping;SHA Yu-ru;LI A-rong;GONG Pu-yang;GU Jian;TAN Rui(College of Pharmacy,Southwest Minzu University,Chengdu 610041,China;School of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610031,China)

机构地区：[1]西南民族大学药学院,四川成都610041 [2]西南交通大学生命科学与工程学院,四川成都610031

出　　处：《中国中药杂志》2022年第8期2038-2048,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(82174083);四川省重点研发项目(20ZDYF3291);四川省科技厅项目(2019YFS0157);教育部重点实验室开放课题(KF2020009);西南民族大学中央高校基本科研业务费专项(2020NZD06)。

摘　　要：探究藏族药二十五味松石丸(Ershiwuwei Songshi Pills,ESP)对非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)小鼠肠道菌群的影响。将48只雄性C57BL/6小鼠随机分为空白组(control),模型组(MCD),藏族药二十五味松石丸高(ESP 0.8 g·kg^(-1))、中(ESP 0.4 g·kg^(-1))、低(ESP 0.2 g·kg^(-1))剂量组和吡格列酮组(pioglitazone,PGZ 10 mg·kg^(-1)),每组8只。空白组普通饲料喂养,其余5组蛋氨酸-胆碱缺乏(methionine-choline-deficient,MCD)饲料喂养5周构建非酒精性脂肪性肝炎(NASH)模型,造模的同时灌胃(ig)给予相应的药物进行干预;实验过程中记录小鼠体质量、日饮水量及日摄食量的变化;实验结束后收集肝组织,苏木精-伊红染色(HE)染色观察肝脏病理变化,油红O染色观察肝脏脂肪堆积情况,检测血清中谷草转氨酶(AST)、谷丙转氨酶(ALT)和肝组织中甘油三酯(TG)的含量;采用16S rRNA高通量测序技术检测小鼠肠道菌群的变化。结果显示,与模型组相比,二十五味松石丸高、中、低剂量组和吡格列酮组血清中的AST、ALT和肝组织中TG含量显著降低,对肝细胞脂肪变性和肝脏中脂肪堆积均有改善作用。16S rRNA测序结果显示,模型组肠道菌群丰度和多样性指数降低,二十五味松石丸组小鼠肠道菌群的丰度和多样性指数升高,并且在门水平上降低了厚壁菌门Firmicutes与拟杆菌门Bacteroidetes的比例;在改变肠道菌群组成方面,降低了肠道菌群中Erysipelotrichia和Faecalibaculum的丰度,提高了Desulfovibrionaceae、Rikenellaceae、Lachnospiraceae和Ruminococcaceae的丰度。该研究发现藏族药二十五味松石丸对MCD饮食诱导的NASH具有保护作用,其作用机制可能是通过调节肠道菌群的变化,改变肠道菌群的组成,抑制肠道菌群的失调,进而改善NASH。This study aimed to investigate the effect of Tibetan medicine Ershiwuwei Songshi Pills(ESP)on the intestinal flora of non-alcoholic steatohepatitis(NASH)mice.Forty-eight male C57 BL/6 mice were randomly divided into the control group,model(methionine-choline-deficient,MCD)group,high-(0.8 g·kg^(-1)),medium-(0.4 g·kg^(-1)),and low-dose(0.2 g·kg^(-1))ESP groups,and pioglitazone(PGZ,10 mg·kg^(-1))group,with eight mice in each group.Mice in the control group were fed with normal diet,while those in the remaining five groups with MCD diet for five weeks for inducing NASH.During modeling,they were gavaged with the corresponding drugs.The changes in body mass,daily water intake,and daily food intake were recorded.At the end of the experiment,the liver tissues were collected and stained with hematoxylin-eosin(HE)for observing the pathological changes,followed by oil red O staining for observing fat accumulation in the liver.The levels of serum aspartate aminotransferase(AST)and alanine aminotransferase(ALT)and triglyceride(TG)in liver tissue were measured.The changes in intestinal flora of mice were determined using 16 S rRNA high-throughput sequencing technology.The results showed that compared with the model group,the high-,medium-and low-dose ESP groups and the PGZ group exhibited significantly lowered AST and ALT in serum and TG in liver tissues and alleviated hepatocellular steatosis and fat accumulation in the liver.As demonstrated by 16 S rRNA sequencing,the abundance index and diversity of intestinal flora decreased in the model group,while those increased in the ESP groups.Besides,the Firmicutes to Bacteroidetes ratio decreased at the phylum level.In the alteration of the composition of intestinal flora,ESP reduced the abundance of Erysipelotrichia and Faecalibaculum but increased the abundance of Desulfovibrionaceae,Rikenellaceae,Lachnospiraceae,and Ruminococcaceae.This study has revealed that ESP has a protective effect against NASH induced by MCD diet,which may be related to its regulation of the changes

关 键 词：藏族药二十五味松石丸 非酒精性脂肪性肝炎(NASH) 蛋氨酸-胆碱缺乏(MCD) 肠道菌群 

分 类 号：R29[医药卫生—民族医学]