病毒性肺炎湿热证小鼠模型的建立及评价  

Establishment and Evaluation of a Mouse Model Simulating Viral Pneumonia with Damp Heat Syndrome

作　　者：张庭瑞 杨上松 向纯 温伟波 李钦 ZHANG Tingrui;YANG Shangsong;XIANG Chun;WEN Weibo;LI Qin(Yunnan University of Traditional Chinese Medicine,Kunming Yunnan 650000,China;Yunnan Key Laboratory of Integrated Traditional Chinese and Western Medicine for Chronic Disease in Prevention and Treatment,Kunming Yunnan 650000,China;Xiantao Traditional Chinese Medicine Hospital,Xiantao Hubei 433099,China)

机构地区：[1]云南中医药大学,云南昆明650000 [2]云南省中西医结合慢病防治重点实验室,云南昆明650000 [3]仙桃市中医医院,湖北仙桃433099

出　　处：《中医药导报》2025年第3期51-59,共9页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家重点研发计划课题(2023YFF0724803);国家自然科学基金项目(8226151028)。

摘　　要：目的:构建并评价病毒性肺炎湿热证小鼠模型。方法:将32只SPF级雄性C57/BL6小鼠随机分为正常组、湿热组、病毒模拟物组、湿热+病毒模拟物组,每组8只。湿热组采用高脂饮食和气候箱湿热暴露14 d的方法构建小鼠湿热证模型,病毒模拟物组采用气管内滴注Poly(I:C)的方法构建小鼠病毒性肺炎肺损伤模型,湿热+病毒模拟物组采用高脂饮食和气候箱湿热暴露14 d叠加气管内滴注Poly(I:C)复合造模法,建立病毒性肺炎湿热证小鼠模型。观测小鼠体质量、食量、饮水量及证候评分变化,分别用酶联免疫法、酶联免疫组织化学染色法、血气分析仪、透射电镜观察血脂代谢(血清HDL-C及LDL-C水平)、胃肠功能(血清MTL、SP水平)、能量代谢(肝组织Na^(+)-K^(+)-ATP酶活性)、水液转运(肺组织AQP1、AQP5、Na^(+)-K^(+)-ATP水平,肺组织细胞线粒体结构)、肺泡-毛细血管屏障(肺组织W/D,BALF中总细胞、总蛋白水平),炎症反应(BALF中TNF-α、IL-1β、IL-6水平)和肺生理功能(动脉血PaO_(2)、PaCO_(2)、FiO_(2)水平),HE染色观察肺组织病理学变化,16SrRNA测序技术观察小鼠肠道菌群。结果:与正常组比较,湿热+病毒模拟物组小鼠的食量、饮水量、体质量下降,湿热证候积分,血清LDL-C、MTL、SP水平,肺组织AQP1、AQP5表达,肺组织W/D、BALF中总细胞数、总蛋白浓度,BALF中TNF-α、IL-1β、IL-6水平,动脉血PaCO_(2)升高(P<0.05或P<0.01);肝、肺组织Na^(+)-K^(+)-ATP酶活性,动脉血PaO_(2)、PaO_(2)/FiO_(2)水平下降(P<0.05或P<0.01);肺组织结构,细胞线粒体结构破坏,肺组织病理损伤评分升高。肠道菌群发生了改变,在属水平主要是拟杆菌、副拟杆菌、屎豆属菌、毛螺菌、NK4A136菌、罗氏菌、NK4A214菌、埃希氏菌相对丰度存在差异,肠道菌群的功能则在需氧化能异养,硝酸盐还原,动物寄生生物或共生体,人类病原体,发酵等方面表现出差异。结论:高脂饮食和气候箱Objective:To construct and evaluate a mouse model of simulated viral pneumonia with damp heat syndrome.Method:Totally 32 SPF grade male C57/BL6 mice were randomly divided into normal group,damp heat group,virus mimic group and damp heat virus mimic group,with 8 mice in each group.The damp heat group constructed a mouse model of damp heat syndrome using a combination of rich and sweet diet and 14 days exposure to humid heat in a climate box.The virus mimetic group constructed a mouse model of simulated viral pneumonia using tracheal instillation of Poly(I∶C).The damp heat virus mimetic group established a mouse model of simulated viral pneumonia damp heat syndrome using a combination of rich and sweet diet and 14 days exposure to humid heat in a climate box through a superimposed gas tube instillation of Poly(I∶C).The changes in body weight,food intake,dietary intake and syndrome scores were observed.Enzyme-linked immunosorbent assay,enzyme-linked immunohistochemistry staining,blood gas analyzer,and transmission electron microscopy were applied to observe lipid metabolism(serum HDL-C and LDL-C levels),gastrointestinal function(serum MTL and SP level),energy metabolism(Na^(+)-K^(+)-ATPase activity in liver tissue),water liquid transport(AQP1,AQP5 and Na^(+)-K^(+)-ATPase levels in lung tissue,mitochondrial structure of lung tissue cells),alveolar capillary barrier(lung tissue W/D,total cell and protein levels in BALF),inflammatory response(TNF-α,IL-1β and IL-6 in BALF),and pulmonary physiological function(arterial blood PaO_(2),PaCO_(2) and FiO_(2) levels).HE staining was used to observe pathological changes in lung tissue,and 16s RNA sequencing technology was used to observe the gut microbiota of mice.Results:Compared with the normal group,the food intake,water intake and weight decreased in damp heat virus mimic group.Damp heat syndrome score,serum LDL-C,MTL and SP levels,lung tissue AQP1 and AQP5 expression,lung tissue W/D,total cell count and total protein concentration in BALF,TNF-α,IL-1βand IL-6 levels

关 键 词：病毒性肺炎 湿热证 病证结合 模型评价 Poly(I:C) 

分 类 号：R2-03[医药卫生—中医学]