基于广州管圆线虫感染导致Balb/c小鼠大脑脱髓鞘的非靶向代谢组学分析  

Untargeted Metabolomics Analysis of Demyelination in the Brain of Balb/c Mice Infected by Angiostrongylus cantonensis

作　　者：牛珍 吴小杰 杨亮 马芷璇 杨君雄 冯英 NIU Zhen;WU Xiaojie;YANG Liang;MA Zhixuan;YANG Junxiong;FENG Ying(School of Medicine,South China University of Technology,Guangzhou 510080,China;School of life science,Sun Yat-Sen University,Guangzhou 510275,China)

机构地区：[1]华南理工大学医学院,广东广州510080 [2]中山大学生命科学学院,广东广州510275

出　　处：《中山大学学报(医学科学版)》2025年第2期293-300,共8页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广州市科技计划项目(202102080053)。

摘　　要：【目的】观察广州管圆线虫(AC)感染导致Balb/c小鼠大脑脱髓鞘症状并分析胼胝体非靶向代谢组的变化,探讨造成脱髓鞘的机制。【方法】将Balb/c小鼠随机分为对照组(6只)和感染组(6只),感染组每只经灌胃感染30条ACⅢ期幼虫,对照组灌胃等体积生理盐水。感染后第3、6、9、12、15、18、21天检测体质量、视觉及行为学评分评估小鼠神经功能学变化;取感染21 d后脑组织,免疫荧光染色、HE染色及透射电镜观察小鼠大脑髓鞘和视网膜形态学变化;代谢组学检测后,采用火山图和热图分析差异代谢物,倍数变化分布图和条形图分析表征主要代谢物成分,选取差异代谢物进行京都基因与基因组百科全书(KEGG)代谢通路分析和调控网络分析。【结果】Balb/c小鼠在AC感染后第9天出现神经行为学评分下降(P<0.05),第15天出现视觉评分下降(P<0.05),第21天时出现明显的体质量下降(P<0.001)和死亡;同时,透射电镜和免疫荧光染色观察到大脑胼胝体区域髓鞘出现明显损伤,少突胶质细胞明显减少(P<0.001),HE病理染色观察到视网膜节细胞出现明显破坏。代谢组学分析表明,所有差异代谢物中,甘油磷脂(GP)类代谢物占比最高,类固醇(ST)类和鞘脂(SP)类代谢物占比相对较少,其中胆固醇酯CE(20:2)表现为显著上调(P<0.001),而磷脂酰甲醇PMeOH(18:0_18:1)表现为显著下调(P<0.01);KEGG富集分析和调控网络分析表明,差异代谢物主要富集于类固醇生物合成、胆汁分泌、胆固醇代谢等代谢通路,参与鞘脂代谢、神经信号调控、甘油磷脂代谢等关键代谢通路。【结论】AC感染通过多途径影响小鼠的代谢状态,改变了与髓鞘形成和稳定密切相关的代谢物水平。脱髓鞘可能与这些关键代谢通路的紊乱密切相关,尤其是胆固醇代谢和鞘脂代谢的紊乱,可能在脱髓鞘的发生中发挥了核心作用。此外,磷脂代谢的改变和神经信号的异常�【Objective】To investigate the demyelination induced by Angiostrongylus cantonensis(AC)infection in the brain of Balb/c mice and analyze the untargeted metabolomic changes in the corpus callosum,aiming to elucidate the underlying mechanisms【.Methods】Balb/c mice were randomly assigned to a control group(n=6)and an infection group(n=6).The infection group was orally administered 30 third-stage larvae of AC,while the control group received an equal volume of saline.Body weight,visual function,and behavioral scores were measured on post-infection 3,6,9,12,15,18,and 21 days to assess neurological alterations.After 21 days,brain tissues were harvested for immunofluorescence staining,he matoxylin-eosin(HE)staining,and transmission electron microscopy to examine morphological changes in brain myelin and retina.Metabolomics analysis was performed,and differential metabolites were identified using volcano plots and heatmaps.The distribution of fold changes and bar charts were used to profile the key metabolites.These differential metabolites were then subjected to Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis and regulatory network analysis.【Results】On the 9 th day after AC infection,Balb/c mice showed a decline in neurological behavioral scores(P<0.05).By day 15,visual scores decreased(P<0.05),and by day 21,significant weight loss(P<0.001)and mortality were observed.Concurrently,transmission electron microscopy and immunofluorescence staining revealed significant myelin damage in the corpus callosum and a marked reduction in oligodendrocytes(P<0.001).HE staining showed severe retinal ganglion cell damage.Metabolomic analysis revealed that glycerophospholipids were the most abundant differential metabolites,with steroids and sphingolipids being relatively less abundant.Cholesteryl ester CE(20:2)was significantly upregulated(P<0.001),while phosphatidylmethanol(18:0_18:1)was significantly downregulated(P<0.01).KEGG enrichment and regulatory network analyses demonstrated that the differen

关 键 词：广州管圆线虫 代谢组学 脱髓鞘 胆固醇 磷脂 

分 类 号：R392[医药卫生—免疫学]