利用基质辅助激光解吸电离质谱成像技术研究斑马鱼响应富营养化污染水质内源性脂质分子变化  

MALDI-MSI Analysis of Endogenous Lipid Changes in Zebrafish Exposures to Water Eutrophication

作　　者：张雅文 秦亮 陈路路 王晓东 ZHANG Ya-wen;QIN Liang;CHEN Lu-lu;WANG Xiao-dong(College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China;Centre for Imaging & Systems Biology, Minzu University of China, Beijing 100081, China)

机构地区：[1]中央民族大学生命与环境科学学院,北京100081 [2]生物成像与系统生物学研究中心,北京100081

出　　处：《科学技术与工程》2021年第14期5658-5667,共10页Science Technology and Engineering

基　　金：国家自然科学基金(31770384,21605164);中央民族大学一流大学一流学科项目基金(Yldxxk201819);中华人民共和国国家民族事务委员会中青年英才培养计划基金(10301-0190040110)。

摘　　要：斑马鱼作为一种常见的模式物种,体内易富集水中污染物,常用于农药、有机物等污染水质监测与评估,但其响应富营养化水质污染的分子机制尚不明确。基质辅助激光解吸电离质谱成像(matrix assisted laser desorption ionization mass spectrometry imaging,MALDI-MSI)能有效检测和反映分子空间分布及原位含量信息,已被广泛应用于动植物学、基础医学、药学、微生物等诸多研究领域。采用MALDI-MSI技术,针对富营养化污染水质环境下斑马鱼内源性脂质代谢分子开展原位分析表征;利用多元统计学方法和相关性分析整合数据;进一步挖掘有效关键信息,旨在从脂质分子层面上初步揭示斑马鱼响应富营养化污染水质的相关分子变化规律。研究结果显示:在m/z 500~1000的质量检测范围内,富营养化水质处理组和未处理组斑马鱼有超过50%原位检测的脂质分子存在明显表达与空间分布差异。主成分分析(principal component analysis,PCA)、偏最小二乘法判别分析(partial least squares discriminant analysis,PLS-DA)及皮尔逊相关聚类热图分析(pearson correlation cluster heatmap analysis)进一步证实了上述差异结果,且统计发现磷脂分子的表达受富营养化污染水质的干扰尤为突出,主要包括PC(32∶0)、PC(34∶1)、PC(34∶2)、PC(36∶0)、PC(38∶6)、PC(40∶6)、PE(36∶8)、PE(36∶0)、PE(38∶0)、PE(40∶9)共10种脂质分子。研究结果为进一步深度探究斑马鱼响应富营养化污染水质体内脂质代谢分子机制奠定了一定的实验基础,也证实了MALDI-MSI作为一种新型分子影像技术应用于生态/环境毒理学监测、评估研究的潜在价值。Zebrafish,as a model animal,is commonly-used to monitor and evaluate the quality of polluted water,such as pesticide and/or organic matter pollution,due to its excellent bio-enrichment property.However,the molecular mechanism of zebrafish in response to the water eutrophication is still unclear.Matrix assisted laser desorption ionization-mass spectrometry imaging(MALDI-MSI)is an emerging molecular-imaging technology that enables determination of the spatial distributions and the relative abundances of diverse endogenous compounds in tissues.Thus far,MSI has been applied to diverse areas of biology,including zoology,basic medical science,pharmaceutical science,microbiology,and plant science.MALDI-MSI technology was used to in-situ detection and imaging of endogenous lipid changes between normal and eutrophication exposed zebrafishes.Multivariate statistical methods and correlation analysis were also used for data processing to screen the key lipid entities highly associated with eutrophication.The results reveal that more than 50%lipids detected in the mass range of m/z 500~1000 show significant changes in expression and spatial distribution when zebrafish expose to eutrophication.Principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA)and Pearson correlation cluster heatmap analysis also prove above mentioned results and reveal ten phospholipids,including PC(32∶0),PC(34∶1),PC(34∶2),PC(36∶0),PC(38∶6),PC(40∶6),PE(36∶8),PE(36∶0),PE(38∶0),and PE(40∶9),to be as the key lipid entities highly associated with eutrophication.The results are helpful for further exploring the molecular mechanism of lipid metabolism in zebrafish in response to eutrophication and also enables the continued progression of MALDI-MSI as an emerging,reliable,and rapid ecotoxicological tool for assessing contaminant toxicity.

关 键 词：斑马鱼 富营养化 基质辅助激光解吸电离质谱成像 原位检测 脂质分子 

分 类 号：O657.63[理学—分析化学]