基于hACE2转导建立SARS-CoV-2 spike假病毒感染病证结合的小鼠病毒肺炎模型

Establishment of SARS-CoV-2 Spike Pseudovirus Infected Mice Pneumonia Model Based on hACE2 Transduction

目的建立与评价一种安全、经济、有效的病证结合小鼠新冠病毒感染肺炎模型。方法30只KM种小鼠随机分为空白组、对照组及实验1组、实验2组、实验3组,每组各6只。每天把KM小鼠置于模拟寒湿环境中4 h,将表达hACE2的重组腺相关病毒(pAAV-hACE2)采用改良悬挂法气管内给药转导至肺组织,继以新型冠状病毒(SARS-CoV-2)spike假病毒采用相同的气管内给药方式造模。观察各组小鼠一般情况,检测体重变化、肺指数、血清炎性因子及肺部病理变化。结果免疫组化法显示h ACE2在小鼠肺组织中有效表达;在寒湿环境中给予SARS-CoV-2 spike假病毒后,实验组小鼠表现出类似疫毒犯肺证候的体征;且同样喂养条件和时间下,与空白组和对照组小鼠比较,实验组小鼠出现体重下降;实验1、2、3组小鼠出现肺指数显著增大(P<0.05或P<0.01),血清炎性因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)显著升高(P<0.01),肺组织病理改变明显。结论本研究成功建立了一种安全、经济、有效的病证结合小鼠新冠病毒感染肺炎模型。

Objective:To establishment and evaluate a safe,economical and effective disease syndrome model of mouse COVID-19 infected pneumonia.Methods:30 KM mice were randomly divided into the blank group,control group,experimental 1,2,3 groups,with 6 mice in each group.KM mice were placed in a simulated cold and humid environment for 4 hours every day,and the recombinant adeno-associated virus expressing hACE2 was transferred into the lung tissue of KM mice by the improved intratracheal administration method,and then the model was made by the same intratracheal administration method of SARS-CoV-2 spike pseudovirus.The general situation of mice in each group was observed,and the changes of body weight,lung index,serum inflammatory factors and lung pathology were detected.Results:Immunohistochemistry showed that hACE2 was effectively expressed in mouse lung tissue;the mice in the experimental group showed signs similar to the lung syndrome caused by epidemic virus after being given SARS-CoV-2 spike pseudovirus in cold and wet environment;under the same feeding conditions and time,compared with the blank group and the control group,the experimental group showed weight loss;the lung index of mice in groups 1,2 and 3 increased significantly(P<0.05 or P<0.01),the levels of serum inflammatory factor IL-1β,IL-6,IL-8,TNF-αwere significantly increased(P<0.01),and the pathological changes of lung tissue were obvious.Conclusion:This study has successfully established a safe,economical and effective model of mouse COVID-19 infection pneumonia combined with disease and syndrome.