伏湿小鼠感染人冠状病毒HCov-229E后免疫功能状态变化  

Experimental Study on Changes in Immune Function Status of Latent Dampness Mice Infected with Human Coronavirus HCov-229E

作　　者：柯宏霞 秦聖乐 许琪华 王玉涛[1] 王新华[1,2] KE Hong-xia;QIN Sheng-le;XU Qi-hua;WANG Yu-tao;WANG Xin-hua(State Key Laboratory of Respiratory Disease,The First Affiliated Hospital of Guangzhou Medical University,Guangzhou,510182;Institute of Inteqration of Traditional and Western Medicine of Guangzhou Medical University,Guangzhou,510000;The Second Clinical Medical College of Guangzhou University of Chinese Medicine(Guangdong Provincial Hospital of Traditional Chinese Medicine),Guangzhou,510120;Artemisinin Research Center,Guangzhou University of Chinese Medicine,Guangzhou,510006)

机构地区：[1]广州医科大学附属第一医院呼吸疾病全国重点实验室,广州510182 [2]广州医科大学中西医结合研究所,广州510000 [3]广州中医药大学第二临床医学院(广东省中医院),广州510120 [4]广州中医药大学青蒿研究中心,广州510006

出　　处：《中国中西医结合杂志》2024年第12期1496-1502,共7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 通过伏湿小鼠模型感染人冠状病毒HCov-229E,探索伏湿状态下病毒感染机体免疫功能的改变。方法 将24只BALB/c小鼠按随机数字表法分为正常组、伏湿组、感染组、伏湿感染组,每组6只;伏湿组和伏湿感染组构建伏湿小鼠模型,感染组和伏湿感染组造模第22天给予10^(-4)HCoV-229E半数组织培养感染剂量(TCID_(50))滴鼻,正常组和伏湿组以磷酸盐缓冲液(PBS)代替。观察肺组织病理;测量脾指数、胸腺指数;检测外周血白细胞(WBC)、淋巴细胞(Lym)、单核细胞(Mono)、中性粒细胞(NE)、红细胞分布宽度变异系数(RDW-CV)、红细胞分布宽度标准差(RDW-SD);Luminex液相芯片法观察血清中趋化因子配体5(CCL5/RANTES)、巨噬细胞炎症蛋白1β(MIP-1β)、干扰素诱导蛋白10 (IP-10)炎症因子在蛋白水平的表达;免疫组化检测肺组织CD4^(+)、CD8^(+)的表达;流式细胞术检测肺CD4+、CD8+T细胞、自然杀伤(NK)细胞的表达。结果 正常组以及伏湿组小鼠肺组织肺泡结构完整,腔内无明显渗出;感染组小鼠肺间质、肺泡腔内出现较多的组织细胞和中性粒细胞浸润,间质小血管周围出现淋巴细胞浸润,伏湿感染组肺泡炎症浸润现象较感染组加重。与正常组比较,伏湿组胸腺指数、脾指数、外周血WBC、Lym、Mono、NE、肺部CD4+表达、CD4+T细胞比例降低,血清RANTES、MIP-1β、IP-10水平升高(P<0.05,P<0.01)。与感染组比较,伏湿感染组胸腺指数和脾指数降低,外周血RDW-CV、RDW-SD、肺部CD4^(+)、CD8^(+)表达、CD4^(+)、CD8^(+)T细胞比例、血清RANTES、MIP-1β、IP-10水平升高(P<0.05,P<0.01)。结论 伏湿小鼠感染人冠状病毒HCov-229E后表现为免疫失衡状态,免疫炎症损伤更严重。Objective To explore the changes in immune function in virus-infected organisms in the latent dampness(LD)state,by establishing a latent dampness mice model infected with human coronavirus HCov-229E.Methods Twenty-four BALB/c mice were randomly divided into four groups:normal group,LD group,infected group,LD-infected group,6 mice in each group.The infected and LD-infected groups were administered half of the tissue culture infectious dose(TCID_(50))of 10^(-4)HCoV-229E via nasal drip on the 22nd day of modeling,while the normal group and LD group received PBS.Lung histopathology was observed,and the splenic index and thymic index were measured.Blood samples were collected to test white blood cell(WBC),lymphocytes(Lym),monocytes(Mono),neutrophils(NE),the coefficient of variation of red blood cell distribution width(RDW-CV),and the standard deviation of red blood cell distribution width(RDW-SD).Luminex liquid chromatography was performed to observe the expression of chemokine cc-motif ligand5(CCL5/RANTES),macrophage inflammatory protein 1β(MIP-1β),interferon inducible protein 10(IP-10)inflammation factors in serum at the protein level.The expression of CD4^(+)and CD8^(+)in lung tissue was detected by immunohistochemistry,and flow cytometry was used to detect the expression of lung CD4+,CD8+T cells and NK cells.Results The alveolar structure of lung tissue was intact in both the normal group and the LD group,and there was no obvious exudation in the cavity.There were more histocyte and neutrophil infiltration in the lung interstitial and alveolar cavities in the infected group,and lymphocyte infiltration around the small blood vessels in the interstitial.The alveolar inflammatory infiltration was more severe in the LD-infected group than in the infected group.Compared with normal group,thymic index,splenic index,WBC,Lym,Mono,NE,lung CD4^(+)expression,CD4^(+)T cell ratio decreased,serum RANTES,MIP-1β,IP-10 levels increased in LD group(P<0.05,P<0.01).Compared with the infected group,thymic index and splenic index de

关 键 词：伏湿 人冠状病毒HCoV-229E 免疫功能 中医证型 中西医结合 

分 类 号：R511[医药卫生—内科学]