iNKT细胞过继转移对哮喘小鼠肺部免疫细胞-神经元相互作用的研究  

Study of immune cell-neuron interactions in the lungs of asthma mice by adoptive transfer of iNKT cells

作　　者：陈亚宾 张雪丽[1] 李宏贵[1] 李燕维 梁俊杰 吉训超[1] 许华[1] CHEN Ya-bin;ZHANG Xue-li;LI Hong-gui;LI Yan-wei;LIANG Jun-jie;JI Xun-chao;XU Hua(Department of Pediatrics,the First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong,China)

机构地区：[1]广州中医药大学第一附属医院儿科,广东广州510405

出　　处：《川北医学院学报》2024年第12期1585-1590,1603,共7页Journal of North Sichuan Medical College

基　　金：国家自然科学基金面上项目(81373686);广东省名中医传承工作室建设项目[粤中医办涵(2023)108号];广东省广州市科技计划项目(2023A04J1172)。

摘　　要：目的:观察iNKT细胞过继转移对哮喘小鼠肺部免疫细胞与神经元相互作用的影响,探讨其在哮喘免疫调节中的潜在机制。方法:选取30只雌性BALB/c小鼠作为研究对象,随机分为对照组、哮喘组和iNKT细胞转移组。对照组接受等量的生理盐水注射,不进行OVA致敏和激发,也不进行iNKT细胞转移;哮喘组按照标准的OVA致敏和激发方案建立哮喘模型,不进行iNKT细胞转移;iNKT细胞转移组在建立哮喘模型后通过尾静脉注射iNKT细胞。模型建立通过HE染色和显微镜观察肺组织的炎症细胞浸润程度进行验证。通过流式细胞术分析T细胞、B细胞和巨噬细胞等肺部免疫细胞群体水平的变化。免疫荧光染色用于检测肺组织神经元标志物(NeuN、ChAT)的表达。ELISA用于检测肺部组织中炎症因子(TNF-α、IL-6、IL-13)和神经递质(乙酰胆碱、去甲肾上腺素)水平。结果:MTT结果显示,与对照组相比,哮喘组的细胞存活率明显更低(P<0.05),iNKT过继转移后,细胞存活率较哮喘组升高(P<0.05);流式细胞术分析显示,与哮喘组相比,iNKT细胞转移组小鼠肺部T细胞和巨噬细胞的比例增加(P<0.05),B细胞比例降低(P<0.05)。免疫荧光染色结果显示,iNKT细胞转移组小鼠肺组织中神经元标志物NeuN和ChAT的表达水平高于哮喘组(P<0.05)。ELISA结果显示,iNKT细胞转移组小鼠的炎症因子TNF-α、IL-6和IL-13的水平显著低于哮喘组(P<0.05),iNKT细胞过继转移组小鼠肺组织中的乙酰胆碱和去甲肾上腺素水平显著高于哮喘组(P<0.05)。结论:过继转移iNKT细胞能够调节哮喘小鼠肺部免疫细胞的组成,降低炎症因子水平,显示出潜在的治疗哮喘的作用。Objective:To investigate the effect of adoptive transfer of iNKT cells on the interaction between lung immune cells and neurons in asthma mice,and to reveal its potential mechanism in asthma immune regulation.Methods:A total of 30 female BALB/c mice were randomly divided into three groups:control group,asthma group and iNKT cell transfer group.The control group received the same amount of normal saline injection,no OVA sensitization and provocation,and no iNKT cell transfer.The asthma group established an asthma model according to the standard OVA sensitization and provocation protocol,and no iNKT cell transfer was performed.The iNKT cell transfer group was injected with iNKT cells through the tail vein after the asthma model was established.The model was successfully established and verified by HE staining and microscopy to observe the degree of inflammatory cell infiltration in lung tissue.Changes in lung immune cell populations such as T cells,B cells,and macrophages were analyzed by flow cytometry.Immunofluorescence staining was used to detect the expression of neuronal markers(NeuN,ChAT)in lung tissue.ELISA was used to detect the levels of inflammatory factors(TNF-α,IL-6,IL-13)and neurotransmitters(acetylcholine,norepinephrine)in lung tissue.Results:MTT results showed that compared with the control group,the cell survival rate in the asthma group was significantly lower than that in the control group(P<0.05).After adoptive transfer of iNKT,the cell survival rate was higher in the asthma group(P<0.05).Flow cytometry Cytometry analysis showed that compared with the asthma group,the proportion of T cells and macrophages in the lungs of mice in the iNKT cell transfer group was increased(P<0.05),while the proportion of B cells was decreased(P<0.05).Immunofluorescence staining results showed that the expression levels of neuronal markers NeuN and ChAT in the lung tissue of mice in the iNKT cell transfer group were higher than those in the asthma group(P<0.05).ELISA results showed that the levels of inflammatory f

关 键 词：INKT细胞 哮喘 免疫细胞 神经元 

分 类 号：R392.12[医药卫生—免疫学]