基于小肠黏膜菌群及肠紧密连接探讨养心通脉方对冠心病血瘀证大鼠的作用机制  

Mechanism of Yangxin Tongmai Prescription for Coronary Heart Disease with Blood Stasis Syndrome Rats Based on Intestinal Mucosal Flora and Intestinal Tight Junction

作　　者：王诗 李杰[1] 刘垠杏 张书萌 陈子君 陈伶利[1,2] WANG Shi;LI Jie;LIU Yinxing;ZHANG Shumeng;CHEN Zijun;CHEN Lingli(Hunan University of Chinese Medicine,Changsha 410208,China;Hunan University of Chinese Medicine Integrative Pathogenic Biology Key Laboratory of Hunan Province,Changsha 410208,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南中医药大学中西医结合病原生物学湖南省重点实验室,湖南长沙410208

出　　处：《中国中医药信息杂志》2022年第8期85-92,共8页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81874375);湖南省自然科学基金(2020JJ4464)。

摘　　要：目的采用16S rRNA测序技术分析冠心病血瘀证大鼠小肠黏膜菌群特点及养心通脉方的作用机制。方法95只SD雄性大鼠随机分为正常组和造模组,造模组采用高脂饲料喂养3个月,结合维生素D3灌胃和异丙肾上腺素皮下注射建立冠心病血瘀证大鼠模型。将成模大鼠随机分为模型组、养心通脉方组(18 g/kg)、阿托伐他汀组(1.8 mg/kg)、正常大鼠粪菌移植组(1 g/kg)、模型大鼠粪菌移植组(1 g/kg)、养心通脉方大鼠粪菌移植组(1 g/kg),每组8只,分别灌胃干预2周。HE染色观察大鼠冠状动脉病理变化,16S rRNA技术对回肠黏膜菌群进行测序分析,透射电镜观察回肠组织超微结构,Western blot检测回肠组织紧密连接蛋白Occludin表达。结果HE染色结果显示,模型组大鼠冠状动脉内膜有纤维帽状突起及大量噬脂细胞沉积,动脉管腔狭窄,冠状动脉粥样硬化斑块百分比升高(P<0.01);养心通脉方组和阿托伐他汀组大鼠冠状动脉病变程度减轻。Alpha多样性分析结果显示,与正常组比较,模型组大鼠肠道菌群丰富度和均匀度降低;与模型组比较,养心通脉方组大鼠肠道菌群丰富度和均匀度升高。Beta多样性分析结果显示,各组大鼠肠道菌群结构有显著差异(q<0.05),养心通脉方组与正常组肠道菌群结构相似,养心通脉方大鼠粪菌移植组和正常大鼠粪菌移植组肠道菌群结构相似。16S rRNA测序分析结果显示,与正常组比较,模型组大鼠肠道菌群厚壁菌门丰度显著降低(P<0.01),变形菌门丰度显著升高(P<0.01);与模型组比较,养心通脉方组大鼠肠道菌群厚壁菌门丰度有升高趋势,变形菌门丰度有降低趋势,但差异无统计学意义(P>0.05)。透射电镜结果显示,与正常组比较,模型组大鼠回肠组织紧密连接减少,桥粒数量减少,养心通脉方组和阿托伐他汀组大鼠回肠组织紧密连接有所改善。Western blot结果显示,与正常组比较,模型组大鼠回肠组�Objective To analyze the characteristics of intestinal mucosal flora in rats with coronary heart disease and blood stasis syndrome and the mechanism of action of Yangxin Tongmai Prescription through 16S rRNA sequencing technology.Methods Totally 95 male SD rats were randomly divided into normal group and modeling group.The modeling group was fed with high-fat diet for 3 months,combined with vitamin D3 gavage and subcutaneous injection of isoproterenol to establish a rat model of coronary heart disease with blood stasis syndrome.The model rats were randomly divided into model group,Yangxin Tongmai Prescription group(18 g/kg),atorvastatin group(1.8 mg/kg),normal rat fecal bacteria transplantation group(1 g/kg),model rat fecal bacteria transplantation group(1 g/kg)and Yangxin Tongmai Prescription fecal bacteria transplantation group(1 g/kg),with 8 rats in each group.They were given intragastric administration for 2 weeks respectively.HE staining was used to observe the pathological changes of coronary arteries;16S rRNA technology was used to sequence and analyze the ileum mucosal flora;transmission electron microscopy was used to observe the ultrastructure of ileal tissue;Western blot was used to detect the expression of tight junction protein Occludin in ileum tissue.Results The results of HE staining showed that the coronary intima of the rats in model group had fibrous cap-like protrusions and a large number of lipophilic deposits,the arterial lumen was narrowed,the percentage of coronary atherosclerotic plaques increased(P<0.01);the degree of coronary artery lesions was alleviated in Yangxin Tongmai Prescription group and atorvastatin group.The results of Alpha diversity analysis showed that compared with the normal group,the richness and uniformity of intestinal flora of the rats in model group decreased;compared with the model group,the richness and uniformity of intestinal flora of the rats in Yangxin Tongmai Prescription group increased.The results of Beta diversity analysis showed that there were significan

关 键 词：肠道菌群 冠心病 血瘀证 养心通脉方 16S rRNA 大鼠 

分 类 号：R285.5[医药卫生—中药学] R256.22[医药卫生—中医学]