摘要
目的通过16S rDNA测序探讨逍遥散调节肠道菌群治疗非酒精性脂肪性肝炎(NASH)模型大鼠的作用机制。方法将30只SD雄性大鼠随机分为正常对照组(Control组,n=10)、模型组(NASH组,n=10)及逍遥散治疗组(XYS组,n=10)。NASH组和XYS组以高脂饲料喂养8周建立NASH大鼠模型,第3周到第6周,每周2次皮下注射40%四氯化碳豆油溶液;XYS组于第2周末开始以逍遥散溶液灌胃直至实验结束。第8周末取各组大鼠肝脏组织采用苏木精-伊红(HE)染色、油红O染色观察肝脏病理组织学改变。采用阿利新蓝-过碘酸雪夫(AB-PAS)染色观察结肠组织杯状细胞黏液层厚度的变化。取各组大鼠结肠内粪便进行16S rDNA高通量测序和微生物多样性分析,以明确NASH组和XYS组大鼠肠道菌群丰度及结构变化。结果与Control组相比,NASH组大鼠肝细胞主要表现为大泡性脂肪变性,伴随气球样变、散在点状坏死和炎细胞浸润,肝小叶结构损坏;大鼠结肠黏膜上皮细胞和杯状细胞萎缩、数量显著减少(P<0.05),黏膜表层细胞黏液的覆盖减少;肠道菌群的α多样性降低,β多样性偏离Control组;厚壁菌门/拟杆菌门比值明显升高;从属水平上看,NASH组抗炎细菌Lachnospiraceae_NK4A136、Roseburia和Blautia的丰度有所减少。与NASH组比较,XYS组大鼠肝组织脂肪变性及炎症浸润情况有不同程度的改善,仅见少许小泡性脂肪变性,炎性细胞浸润减轻;结肠黏膜层上皮细胞和杯状细胞较为圆润饱满;大鼠肠道菌群的α多样性增加,β多样性也逐渐恢复;厚壁菌门/拟杆菌门比值明显降低;从属水平上看,经过逍遥散治疗后,抗炎细菌Lachnospiraceae_NK4A136、Roseburia和Blautia的丰度有所增加。结论逍遥散可以减轻NASH模型大鼠肝细胞的脂肪变性;能够改善NASH模型大鼠肠黏膜的通透性,加强肠道黏膜屏障;并可通过调节肠道菌群多样性及丰度,重塑NASH模型大鼠体内紊乱的肠道菌群。
Objective To explore the mechanism of Xiaoyao Powder in regulating intestinal flora in the treatment of nonalcoholic steatohepatitis(NASH)model rats by using 16SrDNA sequencing.Methods Thirty male SD rats were randomly divided into normal control group(Control,n=10),model group(NASH,n=10)and Xiaoyao powder treatment group(XYS,n=10).NASH group and XYS group were fed high-fat diet for 8weeks to establish NASH rat model.From week 3to week 6,40%carbon tetrachloride soybean oil solution was injected subcutaneously twice a week.The XYS group was given Xiaoyao power solution intragastrically at the end of the second week until the end of the experiment.At the end of 8th week,the liver tissues of each group were taken by hematoxylin-eosin(HE)staining and oil red O staining to observe the histopathological changes of the liver.Alcian blue-periodate Shev(AB-PAS)staining was used to observe the changes of mucous layer thickness of goblet cells in colon tissue.16SrDNA high-throughput sequencing and microbial diversity analysis were performed in colonic feces of rats in each group to determine the intestinal flora abundance and structural changes of rats in NASH group and XYS group.ResultsCompared with the Control group,the hepatocytes in NASH group mainly showed bulla steatosis,accompanied by balloon-like transformation,scattered spot-like necrosis and inflammatory cell infiltration,and damage of hepatic lobular structure.The number of colon mucosal epithelial cells and goblet cells decreased significantly(both P<0.05),and the mucous coverage of mucosal surface cells decreased.Theαdiversity of intestinal flora decreased andβdiversity deviated from Control group.The ratio of Firmicutes to Bacteroidetesincreased significantly.At the secondary level,the abundance of anti-inflammatory bacteria Lachnospiraceae_NK4A136,Roseburia and Blautia decreased in the NASH group.Compared with NASH group,hepatic steatosis and inflammatory infiltration of rats in XYS group were improved to varying degrees,only a little vesicular steatosis wa
作者
孟雅楠
张玉伟
张馨月
刘晋芳
苗宇船
Meng Ya′nan;Zhang Yuwei;Zhang Xinyue(School of Basic Medicine,Shanxi University of Traditional Chinese Medicine,Jinzhong 030619,China;Shanxi Traditional Chinese Medicine Institute,Taiyuan 030012,China)
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2023年第4期517-524,共8页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81470190)
山西省卫生健康委员会科研项目(No.2019089)
山西中医药大学科技创新能力培育计划“基础研究专项”(No.2021-PY-JC-04)。
