环境与职业医学2024，Vol.41Issue(1)：34-40,7.DOI:10.11836/JEOM23225
基于蛋白质组学技术探究无机氟致大脑损伤的机制
Mechanisms of brain damage caused by inorganic fluoride using proteomics-based techniques
摘要
Abstract
[Background]Chronic excessive exposure to fluoride can cause damage to the central nervous system and a certain degree of learning and memory impairment.However,the associated mechanism is not yet clear and further exploration is needed. [Objective]Using 4D unlabelled quantitative proteomics techniques to explore differentially expressed proteins and their potential mechanisms of action in chronic excessive fluoride exposure induced brain injury. [Methods]Twenty-four SPF-grade adult SD rats,half male and half male,were selected and divided into a control group and a fluoride group by random number table method,with 12 rats in each group.Among them,the control group drank tap water(fluorine content<1 mg·L-1),the fluoride group drank sodium fluoride solution(fluorine content 10 mg·L-1),and both groups were fed with ordinary mouse feed(fluoride content<0.6 mg·kg-1).After 180 d of feeding,the SD rats were weighed,and then part of the brain tissue was sampled for pathological examination by hematoxylin-eosin(HE)staining and Nissl staining.The rest of the brain tissue was frozen and stored at-80℃.Three brain tissue samples from each group were randomly selected for proteomics detection.Differentially expressed proteins were screened and subcellular localization analysis was performed,followed by Gene Ontology(GO)function analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis,cluster analysis,and protein-protein interaction analysis.Finally,Western blotting was used to detect the expression levels of key proteins extracted from the brain tissue samples. [Results]After 180 d of feeding,the average weight of the rats in the fluoride group was significantly lower than that in the control group(P<0.05).The brain tissue stained with HE showed no significant morphological changes in the cerebral cortex of the fluoride treated rats,and neuron loss,irregular arrangement of neurons,eosinophilic changes,and cell body pyknosis were observed in the hippocampus.The Nissl staining results showed that the staining of neurons in the cerebral cortex and hippocampus of rats exposed to fluoride decreased(Nissl bodies decreased).The proteomics results showed that a total of 6927 proteins were identified.After screening,206 differentially expressed proteins were obtained between the control group and the fluoride group,including 96 up-regulated proteins and 110 down-regulated proteins.The differential proteins were mainly located in cytoplasm(30.6％),nucleus(27.2％),mitochondria(13.6％),plasma membrane(13.6％),and extracellular domain(11.7％).The GO analysis results showed that differentially expressed proteins mainly par-ticipated in biological processes such as iron ion transport,regulation of dopamine neuron differentiation,and negative regulation of res-piratory burst in inflammatory response,exercised molecular functions such as ferrous binding,iron oxidase activity,and cytokine activi-ty,and were located in the smooth endoplasmic reticulum membrane,fixed components of the membrane,chloride channel complexes,and other cellular components.The KEGG significantly enriched pathways included biosynthesis of secondary metabolites,carbon metabolism,and microbial metabolism in diverse environments.The results of differential protein-protein interaction analysis showed that the highest connectivity was found in glucose-6-phosphate isomerase(Gpi).The expression level of Gpi in the brain tissue of the rats in the fluoride group was lower than that in the control group by Western blotting(P<0.05). [Conclusion]Multiple differentially expressed proteins are present in the brain tissue of rats with chronic fluorosis,and their functions are related to biosynthesis of secondary metabolites,carbon metabolism,and microbial metabolism in diverse environments;Gpi may be involved in cerebral neurological damage caused by chronic overdose fluoride exposure.关键词
慢性氟暴露/大脑损伤/蛋白质组学/差异表达蛋白/葡萄糖-6-磷酸异构酶Key words
chronic fluoride exposure/brain damage/proteomics/differentially expressed protein/glucose-6-phosphate isomerase分类
医药卫生引用本文复制引用
周箫,万雯,姜德文,艾福军,叶凌,刘明海,张艺,刘艳洁..基于蛋白质组学技术探究无机氟致大脑损伤的机制[J].环境与职业医学,2024,41(1):34-40,7.基金项目
国家自然科学基金项目(81960572) （81960572）
国家自然科学基金委员会-贵州省人民政府联合基金子项目(U1812403) （U1812403）
贵州省卫健委科技基金项目(gzwkj2022-217) （gzwkj2022-217）
贵州省教育厅青年科技人才项目(黔教合KY字[2021]183) This study was funded. （黔教合KY字[2021]183）
