摘要
目的观察化瘀祛痰方对ApoE^(-/-)动脉粥样硬化(atherosclerosis,AS)小鼠肠道菌群及黏膜屏障的影响,探讨调控肠道菌群及肠黏膜屏障防治AS的作用机制。方法40只ApoE^(-/-)小鼠随机分为模型组,化瘀祛痰方低、中、高剂量组,10只C57BL/6J小鼠作为正常组。模型组及各治疗组均给予高脂饲料喂养,正常组给予普通饲料喂养。造模成功后化瘀祛痰方不同剂量组分别给予相应的药物灌胃,对照组与模型组给予等量的生理盐水。30 d后16S rDNA分析小鼠肠道菌群变化;HE染色观察肠黏膜屏障病理变化;ELISA法检测血清及肠道内容物中脂多糖(lipopolysaccharide,LPS)的含量;Western blot法及Realtime RT-PCR法检测肠道上皮细胞间紧密连接蛋白ZO-1、Claudin、Occludin1蛋白及mRNA表达。结果与正常组相比,模型组肠道菌群多样性降低,厚壁菌门及放线菌门比例升高,拟杆菌门及变形菌门的比例降低;毛螺菌属(f-Lachnospiraceae)、梭菌目(o-Clastridiales)、梭菌属(c-Clastridia)、脱硫弧菌科(f-Desulfovibrionaceae)、脱硫弧菌目(o-Desulfovibrionales)丰度存在差异。模型组小鼠HE染色显示,肠上皮细胞上皮下间隙增宽,大片绒毛脱落,固有层裸露;血清及盲肠内容物LPS水平升高(P<0.05或P<0.01);肠道上皮细胞间紧密连接蛋白ZO-1、Claudin1、Occludin蛋白及mRNA的表达降低(P<0.05或P<0.01)。化瘀祛痰方干预后,肠上皮细胞上皮下间隙变窄,绒毛脱落减少;血清及盲肠内容物LPS水平显著降低(P<0.05或P<0.01);ZO-1、Claudin1、Occludin蛋白及mRNA的表达升高(P<0.05或P<0.01)。结论化瘀祛痰方可能通过调节ApoE^(-/-)小鼠肠道菌群结构,改善肠道黏膜屏障功能,降低炎性介质LPS释放入血,从而防治动脉粥样硬化。
Objective To observe the effect of Huayu Qutan Prescription(化瘀祛痰方)on intestinal flora and mucosal barrier of ApoE^(-/-)mice and explore the mechanism of regulating intestinal flora and intestinal mucosal barrier in prevention and treatment of atherosclerosis(AS).Methods Forty ApoE^(-/-)mice were randomly divided into model group,Huayu Qutan Prescription low,medium and high dose groups,and 10 c57bl/6j mice were as the normal group.The model group and each treatment group were fed with high fat feed,and the blank group was fed with common feed.After 4 weeks of model making,the different dose groups of Huayu Qutan Prescription were given the corresponding drugs for gastric administration,and the control group and model group were given the same amount of physiological saline.The results showed that 16S rDNA was used to analyze the intestinal flora of mice after 8 weeks.HE staining was used to observe the pathological changes of intestinal barrier.The content of lipopolysaccharide(LPS)in serum and intestinal contents was detected by ELISA.The expressions of close junction proteins ZO-1,Claudin,Occludin1 and mRNA were detected by Western blot and Realtime RT-PCR.Results Compared with those of the normal group,the diversity of intestinal flora in the model group decreased,the proportion of thick walled bacteria and actinomycetes increased,the ratio of pseudobacteria and proteus decreased.The proportions of f-Lachnospiraceae,o-Clastridiales,c-Clarsidia,f-Desulfovibrionaceae,O-Desulfovibrionales had differences in abundance.HE staining showed that the subcutaneous space of the intestinal epithelial cells widened,large villi fell off and the intrinsic layer was exposed.The levels of LPS in serum and cecum content increased significantly(P<0.05 or P<0.01).The expressions of ZO-1,Claudin1 and Occludin were significantly decreased in proteins and mRNA(P<0.05 or P<0.01).After the treatment of Huayu Qutan Prescription,the subcutaneous space of intestinal epithelial cells narrowed and villi shedding decreased.The levels of LPS in serum and cecum content decreased(P<0.05 or P<0.01).The expressions of close junction proteins ZO-1,Claudin1 and occludin were increased(P<0.05 or P<0.01).Conclusion Huayu Qutan Prescription may prevent AS by regulating the intestinal flora structure of ApoE^(-/-)mice,reducing the abundance of pathogenic bacteria,improving the function of intestinal mucosa barrier and reducing the release of inflammatory medium LPS into blood.
作者
郑昊龙
翟亚荣
吴瑶
孟嘉伟
马艺鑫
吕美君
宋囡
隋国媛
杨关林
贾连群
ZHENG Haolong;ZHAI Yarong;WU Yao;MENG Jiawei;MA Yixin;LYU Meijun;SONG Nan;SUI Guoyuan;YANG Guanlin;JIA Lianqun(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Shenyang Tenth People's Hospital,Shenyang 110847,Liaoning,China;Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)
出处
《中华中医药学刊》
CAS
北大核心
2023年第1期81-85,I0028,I0029,共7页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(81974548,82074145)
辽宁省“兴辽人才”计划项目(XLYC1902100)
辽宁省自然科学基金项目(2019-MS-288)。