表没食子儿茶素没食子酸酯抑制铅暴露后NLRP3炎症体介导的神经炎症反应  

Epigallocatechin gallate inhibits lead exposure-induced neuroinflammation mediated by NLRP3 inflammasome

作　　者：冯鉴 王倩 姜雨昕 熊晓锋 于凯伦 杨勋博 芦嘉桥 刘启玲 王迪雅 FENG Jian;WANG Qian;JIANG Yuxin;XIONG Xiaofeng;YU Kailun;YANG Xunbo;LU Jiaqiao;LIU Qiling;WANG Diya(School of Public Health,Shaanxi University of Chinese Medicine,Xianyang,Shaanxi 712046,China;Department of Military Occupational and Environmental Health,Military Preventive Medicine School/Ministry of Education Key Laboratory of Hazard Assessment and Control in Special Operational Environment,Air Force Medical University,Xi'an,Shaanxi 710032,China)

机构地区：[1]陕西中医药大学公共卫生学院,陕西咸阳712046 [2]空军军医大学军事预防医学系军队劳动与环境卫生学教研室/特殊作业环境危害评估与防治教育部重点实验室,陕西西安710032

出　　处：《环境与职业医学》2025年第1期95-102,共8页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金面上项目(82273592);中国科协青年人才托举工程(2018QNRC002)。

摘　　要：[背景]铅(Pb)是一种环境中普遍存在的有毒污染物,有研究表明Pb暴露会诱导小胶质细胞活化,引起神经损伤,加大患阿尔茨海默病等神经退行性疾病的风险。小胶质细胞的活化受核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症体的调控,但如何有效抑制NLRP3炎症体的活化,干预神经炎症的发生发展仍不清楚。[目的]探讨表没食子儿茶素没食子酸酯(EGCG)是否可以抑制Pb暴露后NLRP3炎症体介导的神经炎症。[方法]建立小鼠醋酸铅饮水暴露模型,小鼠分为4组:对照(Con)、EGCG、Pb、EGCG+Pb,每组18只;其中Pb组和EGCG+Pb组给予0.2%醋酸铅饮水,Con和EGCG组正常饮水;EGCG组和EGCG+Pb组给予EGCG灌胃,每周三次,持续一月;其余组采用生理盐水灌胃,灌胃条件相同。小鼠醋酸铅染毒和EGCG灌胃干预同时进行。采用石墨原子吸收光谱法测量小鼠血铅水平;通过Morris水迷宫、新物体识别实验、恐惧条件箱检测小鼠学习记忆水平;采用Western blot、免疫荧光染色法、高尔基染色法来观察神经元的损伤情况;通过免疫荧光染色法观察小胶质细胞活化后的形态变化;采用PCR、Western blot检测白介素(IL)-1β、IL-18、肿瘤坏死因子-α(TNF-α)、iNOS等炎症因子以及NLRP3炎症体相关分子NLRP3、凋亡相关斑点样蛋白(ASC)、半胱氨酸蛋白酶-1(Caspase-1)的mRNA及蛋白的相对表达水平,以此观察Pb暴露对小胶质细胞活化和NLRP3炎症体激活的影响。在EGCG灌胃干预后再次检测上述指标,以探究EGCG干预对NLRP3炎症体活化以及神经炎症发生发展的作用。[结果]相较于Con组,Pb暴露后,小鼠血铅值升高(P <0.05)。Pb组小鼠在Morris水迷宫中的平台象限停留时间缩短,穿台次数也有所减少(P <0.05)。新物体识别实验中,Pb组小鼠辨别指数(DI)和认知指数(RI)下降(P <0.05)。恐惧条件箱实验中,Pb组小鼠的凝滞指数下降(P <0.05)。高尔基染色结果显示,Pb组小鼠树突棘�[Background]Lead(Pb)is an ubiquitous toxic pollutant in the environment,and studies have shown that Pb exposure induces microglia activation,causing neurological damage and increasing the risk of neurodegenerative diseases such as Alzheimer's disease.Microglia ac-tivation is regulated by nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome,but how to effectively inhibit NLRP3 inflammasome activation and prevent neuroinflammation is still unclear.[Objective]To investigate whether epigallocatechin gallate(EGCG)can inhibit Pb-induced neuroinflammation mediated by NLRP3 in-flammasome.[Methods]A mouse model of lead acetate drinking exposure was established,and the mice were divided into 4 groups:control(Con),EGCG,Pb,and EGCG+Pb,with 18 mice in each group.The Pb group and the EGCG+Pb group were given 0.2%lead acetate drinking water,while the Con group and the EGCG group were given normal drinking water.The EGCG group and the EGCG+Pb group were given EGCG via gavage,three times a week for a month.The other groups were given saline via gavage under the same conditions.The lead acetate exposure and EGCG gavage intervention were conducted simultaneously.Blood lead levels were measured by graphite atomic absorption spectroscopy.Learning and memory of the mice were tested using Morris water maze,novel object recognition test,and fear conditioning box.Neuronal damage was observed using Western blot,immunofluorescence staining,and Golgi staining.The morphological changes of microglia activation were observed after immunofluorescence staining.The expression levels of inflammatory factors such as interleukin(IL)-1β,IL-18,tumor necrosis factor-α(TNF-α),and inducible nitric oxide synthase(iNOS),as well as NLRP3 inflammasome-related molecules such as NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),and Caspase-1,were detected by PCR and Western blot to observe the effects of Pb exposure on microglia activation and NLRP3 inflammasome activation.The above indicators were re-exami

关 键 词：表没食子儿茶素没食子酸酯 铅暴露 核苷酸结合寡聚化结构域样受体蛋白3 神经炎症 

分 类 号：R11[医药卫生—公共卫生与预防医学]