从肠道菌群动态变化及炎症因子探讨温病湿热证实质  

Exploring the Essence of Damp-Heat Syndrome of Febrile Disease from the Dynamic Change of IntestinaI lFloraaand Inflammatory Factors

作　　者：纪丽纯 罗子清 马利娟 林樫 赵可伟 林兴栋 JI Lichun;LUO Ziqing;MA Lijuan;LIN Jian;ZHAO Kewei;LIN Xingdong(The Third Clinical Medical School,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Animal Experiment Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The Third Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510378 Guangdong,China)

机构地区：[1]广州中医药大学第三临床医学院,广东广州510405 [2]广州中医药大学实验动物中心,广东广州510405 [3]广州中医药大学第三附属医院,广东广州510378

出　　处：《中药新药与临床药理》2023年第10期1409-1418,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省自然科学基金项目(2020A1515010392);广东省中医药局科研项目(20213008)。

摘　　要：目的基于复合多因素复制湿热证小鼠模型,从肠道菌群动态变化及炎症机制,探讨温病湿热证“湿”“热”实质。方法随机将96只BALB/c雄性小鼠分为8组:对照组、病毒组、湿模组、湿治组、湿粪组、热模组、热治组、热粪组。以湿热环境、高脂饮食、中药和流感病毒滴鼻进行模型复制,治疗组给予中药或粪菌液灌胃,其余小鼠灌胃等体积生理盐水。实时荧光定量聚合酶链式反应(q PCR)检测肺内病毒相对表达量,免疫组化检测结肠组织中水通道蛋白3(AQP3)和AQP4,酶联免疫吸附法(ELISA)检测血清白细胞介素(IL)-1β、干扰素γ(IFN-γ)、IL-4含量,对粪便进行16S rDNA测序。结果与对照组比较,病毒组、湿模组和热模组肺病毒相对表达量明显升高(P<0.05);结肠中AQP3和AQP4表达均不同程度增高或降低(P>0.05);血清均出现不同程度IL-1β、IFN-γ升高及IL-4降低(P<0.05);肠道菌群16S测序发现“湿”“热”模型组差异的主要菌群是乳酸菌、葡萄球菌、拟杆菌等。结论湿模组小鼠肺病毒相对表达量、炎症因子变化程度明显高于热模组,AQP3表达减少,优势菌群中益生菌乳酸菌相对丰度小于热模组,湿热证“湿”“热”模型存在差异。Objective To replicate a mouse model of damp-heat syndrome induced by multiple factors,and to explore the essence of“dampness”and“heat”in the damp-heat syndrome of warm diseases from the dynamic changes of gut microbiota and inflammatory mechanisms.Methods Ninety-six BALB/c male mice were randomly divided into eight groups:control group,virus group,damp model group,damp treatment group,damp feces group,heat model group,heat treatment group,and heat feces group.The mice model was replicated by damp and heat environment,high-fat diet,Chinese medicine and nasal drip of influenza virus.After modeling,the treatment groups were given Chinese medicine or fecal bacteria solution,while the others were given an equal volume of normal saline.The relative expression of the pulmonary virus was detected by real-time fluorescence quantitative PCR.The expressions of AQP3 and AQP4 in colon tissue were determined by immunohistochemistry.The contents of IL-1β,IFN-γ,and IL-4 in serum were determined by ELISA.Fecal samples were determined by the analysis of 16S rDNA sequencing.Results Compared with the control group,the relative expression of the pulmonary virus of mice in the virus,damp model,and heat model groups was significantly increased(P<0.05).The expressions of AQP3 and AQP4 in the colon were increased or decreased to different degrees(P>0.05).The serum levels of IL-1βand IFN-γincreased and IL-4 decreased to different degrees(P<0.05).The 16S sequencing analysis of intestinal flora revealed that the main differences in main intestinal flora between the damp model group and the heat model group were Lactobacllus,Staphylococcus,Bacteroides,etc.Conclusion The relative expressions of lung viruses and inflammatory factors in the damp model group were significantly higher than those in the heat model group,the expression of AQP3 decreased,and the relative abundance of lactobacillus(a type of probiotic)in the dominant bacteria was less than that in the heat model group.There were differences in the damp-and heat-models of

关 键 词：湿热证 肠道菌群 动物模型 粪菌移植 流感 炎症因子 小鼠 

分 类 号：R-332[医药卫生] R285.5