伏湿小鼠模型构建及其对人冠状病毒HCoV-229E易感性研究  

Establishment of Latent dampness Mice Model and Its Susceptability to Coronavirus HCoV-229E

作　　者：秦聖乐 杨子峰[1] 赵瑾[1] 黄婉怡[1] 邓玲珠 王玉涛[1] 王新华[1] QIN Sheng-le;YANG Zi-feng;ZHAO Jin;HUANG Wan-yi;DENG Ling-zhu;WANG Yu-tao;WANG Xin-hua(State Key Laboratory of Respiratory Disease,Guangzhou Institute of Respiratory Health,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510182)

机构地区：[1]广州医科大学附属第一医院,广州呼吸健康研究院,呼吸疾病国家重点实验室,广州510182

出　　处：《中国中西医结合杂志》2022年第11期1348-1355,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.82074311)。

摘　　要：目的构建小鼠伏湿模型,并观察该模型对人冠状病毒HCoV-229E的易感性。方法将30只BALB/c小鼠随机分为正常组、模型组和中药组,每组10只。模型组和中药组采用高脂饮食+湿度(95±5)%环境21 d建立伏湿模型小鼠,中药组15 d起给予给予五指防冠方(11.44 g/kg)灌胃。另取64只小鼠随机分为正常组、正常低滴度[10^(-3)HCoV-229E半数组织培养感染剂量(TCID_(50))]感染组、正常中滴度(10^(-4)TCID_(50))感染组、正常高滴度(10^(-5)TCID_(50))感染组、模型组、伏湿低滴度(10^(-3)TCID_(50))感染组、伏湿中滴度(10^(-4)TCID_(50))感染组、伏湿高滴度(10^(-5)TCID_(50))感染组,每组8只,各组给予相应滴度的HCoV-229E滴鼻,正常组和模型组不予干预。观察小鼠一般情况,测量小鼠体重、肺指数及肺、小肠组织脂多糖(LPS)含量,观察舌组织、肺组织、结肠组织病理,并进行肠道微生物16S rRNA基因测序;检测肺组织炎症因子炎症因子干扰素诱导蛋白10(IP-10)、单核细胞趋化蛋白1(MCP-1)、巨噬细胞炎症蛋白β(MIP-β)、趋化因子配体5(RANTES)mRNA表达水平。结果正常组小鼠被毛光滑,大便成形;模型组小鼠出现被毛潮湿、大便变软,肛周黄秽、饮水量减少的表现,结肠隐窝内有少量中性粒细胞浸润;中药组上述表现有所缓解。与正常组比较,模型组的Shannon、Chao1、Sob指数及拟杆菌门、变形菌门、Alistipes、Bacteroides、Ruminiclostridium_5、Ruminiclostridium_9属相对丰度降低(P<0.05,P<0.01),舌组织角化及放线菌门、Jeotgalicoccus、Corynebacterium_1属相对丰度升高(P<0.01)。正常组和模型组肠道微生物菌落构成及19类功能基因有差异(P<0.05,P<0.01),功能基因涉及糖、辅酶和维生素、氨基酸代谢等。与模型组比较,中药组的Chao1、Sob指数及拟杆菌门、Alistipes、Bacteroides、Ruminiclostridium_5、Ruminiclostridium_9属相对丰度升高(P<0.05,P<0.01),舌组织角化及Jeotgalicoccus、CorynebacObjective To establish Latent dampness mouse model and observe its susceptibility to human coronavirus HCoV-229E.Methods Totally 30 BALB/c mice were randomly divided into normal group,model group and Chinese medicine(CM)group(n=10).Model group and CM group were fed by high fat diet and exposure to high humidity(95±5)%21 d to establish Latent dampness mouse model.CM group was administered intagastrically Wuzhi Fangguan Formula(WZFGF)from 15 d at a dose of 11.44 g/kg.Another 64BALB/c mice were randomly divided into normal group,normal low titer[10^(-3)HCoV-229E 50%tissue culture infective dose(TCID_(50))]infection group,normal medium titer(10^(-4)TCID_(50))infection group,normal high titer(10^(-5)TCID_(50))infection group,model group,latent dampness low titer(10^(-3)TCID_(50))infection group,latent dampness medium titer(10^(-4)TCID_(50))infection group and latent dampness high titer(10^(-5)TCID_(50))infection group(n=8).Each group was given the corresponding titer of HCoV-229E by nasal drops,the normal group and the model group were not intervened.The state of mice was observed,body weight,lung index,lung and small intestine lipopolysaccharide(LPS)level were measured.The histopathology of tongue,lung and colon tissues of mice was observed The gut microbes of mice were investigated by 16S rRNA gene sequencing.The mRNA expression of inflammatory factors interferon-inducible protein 10(IP-10),monocyte chemoattractant protein-1(MCP-1)、macrophage inflammatory proteinβ(MIP-β),chemokine cc-motif ligand5(RANTES)in lung tissues were detected.Results Mice in the normal group had smooth hair and formed stools.Mice in model group showed the performance of moist hair,soft stool,yellow feces around anus,and less water drinking,a bit neutrophil infiltration appeared in the crypt of colon tissues.The changes of latent dampness mouse model group were attenuated by CM treatment.Compared with the normal group,Shannon,Chao1 and Sob index,abundances of phylum Bacteroidetes and Proteobacteria,genus Alistipes,Bacteroides,Ruminiclost

关 键 词：伏湿模型 人冠状病毒HCoV-229E 肠道微生物 易感性 炎症因子 

分 类 号：R-332[医药卫生] R255