基于GC-MS研究OPFRs对PC12细胞代谢的影响

摘要:

基于气相色谱质谱(gas chromatography-mass spectrometry,GC-MS)研究了4种有机磷阻燃剂(organophosphate flame retardants,OPFRs)对神经细胞PC12代谢的影响.首先将PC12细胞分别暴露于4种不同浓度的阻燃剂中,利用噻唑蓝(methyl thiazolyl tetrazolium,MTT)比色法选出对细胞存活率影响较小的浓度.然后在该浓度下培养细胞并提取代谢物,利用GC-MS对细胞内的代谢物进行非靶向代谢组学分析.接着采用SIMCA软件对定量后的代谢数据进行OPLS-DA分析,找出细胞中受到OPFRs影响的关键代谢物,并利用生物信息学方法分析这些关键代谢物所涉及的重要代谢通路.该研究共鉴定出PC12细胞中的38种小分子代谢物.经过多元统计分析发现4种OPFRs均影响PC12细胞的代谢表型.OPFRs在100μmol/L和1000μmol/L浓度下,对PC12细胞表现出显著的细胞毒性.其中糖代谢和氨基酸代谢是受影响的主要代谢通路.

This research explored the effects of four organophosphorus flame retardants(OPFRs)on the metabolism of nerve cells(PC12)bases on gas chromatography-mass spectrometry(GC-MS).Firstly,PC12 cells were exposed to various concentrations of four flame retardants,and the concentration that had minimal effect on cell viability was determined by methyl thiazolyl tetrazolium(MTT)colorimetry.Then,the cells were cultured at this concentration,and metabolites were extracted and analyzed by using GC-MS for non-targeted metabolomics.Subsequently,SIMCA software was utilized to perform OPLS-DA analysis on the quantitative metabolic data to discover the significant metabolites impacted by OPFRs in cells.Finally,bioinformatics methods were employed to dissect the pertinent metabolic pathways linked to these key metabolites.A total of 38 small-molecule metabolites were identified in PC12 cells.Multivariate statistical analysis showed that all the four OPFRs affected the metabolic phenotype in PC12 cells.OPFRs showed significant cytotoxicity against PC12 cells at 100μmol/L and 1000μmol/L concentrations.Carbohydrate metabolism and amino acid metabolism were mainly affected in cells.

作者:

孙梦瑶 赵亚菲 王少敏 刘宏民

Sun Mengyao;Zhao Yafei;Wang Shaomin;Liu Hongmin(School of Ecology and Environment,Zhengzhou University,Zhengzhou 450001,China;College of Chemistry,Zhengzhou University,Zhengzhou 450001,China;Collaborative Innovation Center of New Drug Research and Safety Evaluation of Henan Province,Zhengzhou University,Zhengzhou 450001,China)

机构地区:

郑州大学生态与环境学院 郑州大学化学学院 郑州大学新药创制与药物安全性评价河南省协同创新中心

引用本文:

《betway官方app 学报（自然科学版）》 CAS  2024年第2期96-103,I0005,共9页

Journal of Henan Normal University(Natural Science Edition)

基金:

国家自然科学基金(21772180).

关键词:

有机磷阻燃剂 代谢组学 气相色谱质谱 PC12细胞

organophosphate flame retardants metabolomics GC-MS PC12 cells

分类号:

R114 [医药卫生—卫生毒理学] 

基于GC-MS研究OPFRs对PC12细胞代谢的影响.pdf