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基质辅助激光解吸电离成像技术对斑马鱼肝脏中全氟辛酸积聚及其对磷脂影响的可视化研究。

Visualization of PFOA accumulation and its effects on phospholipid in zebrafish liver by MALDI Imaging.

机构信息

State Key Laboratory of Environmental Criteria and Risk Assessment, National Engineering Laboratory for Lake Pollution Control and Ecological Restoration, State Environmental Protection Key Laboratory for Lake Pollution Control, Chinese Research Academy of Environmental Sciences, Beijing, 100012, China.

Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, 361021, China.

出版信息

Anal Bioanal Chem. 2024 Apr;416(10):2493-2501. doi: 10.1007/s00216-024-05214-y. Epub 2024 Mar 7.

Abstract

Exposure to poly- and perfluoroalkyl substances (PFASs) can result in bioaccumulation. Initial findings suggested that PFASs could accumulate in tissues rich in both phospholipids and proteins. However, our current understanding is limited to the average concentration of PFASs or phospholipid content across entire tissue matrices, leaving unresolved the spatial variations of lipid metabolism associated with PFOA in zebrafish tissue. To address gap, we developed a novel methodology for concurrent spatial profiling of perfluorooctanoic acid (PFOA) and individual phospholipids within zebrafish hepatic tissue sections, utilizing matrix-assisted laser desorption/ionization time of flight imaging mass spectrometry (MALDI-TOF-MSI). 5-diaminonapthalene (DAN) matrix and laser sensitivity of 50.0 were optimized for PFOA detection in MALDI-TOF-MSI analysis with high spatial resolution (25 μm). PFOA was observed to accumulate within zebrafish liver tissue. H&E staining results corroborating the damage inflicted by PFOA accumulation, consistent with MALDI MSI results. Significant up-regulation of 15 phospholipid species was observed in zebrafish groups exposed to PFOA, with these phospholipid demonstrating varied spatial distribution within the same tissue. Furthermore, co-localized imaging of distinct phospholipids and PFOA within identical tissue sections suggested there could be two distinct potential interactions between PFOA and phospholipids, which required further investigation. The MALDI-TOF-IMS provides a new tool to explore in situ spatial distributions and variations of the endogenous metabolites for the health risk assessment and ecotoxicology of emerging environmental pollutants.

摘要

接触多氟和全氟烷基物质 (PFASs) 可能导致生物蓄积。最初的研究结果表明,PFASs 可能会在富含磷脂和蛋白质的组织中蓄积。然而,我们目前的认识仅限于整个组织基质中 PFASs 或磷脂含量的平均浓度,因此仍然不清楚与全氟辛酸 (PFOA) 相关的脂质代谢在斑马鱼组织中的空间变化。为了解决这一差距,我们开发了一种新的方法,用于在斑马鱼肝组织切片中同时对全氟辛酸 (PFOA) 和单个磷脂进行空间分析,该方法利用基质辅助激光解吸/电离飞行时间成像质谱 (MALDI-TOF-MSI)。优化了 5-二氨基萘 (DAN) 基质和激光灵敏度,以实现 MALDI-TOF-MSI 分析中具有高空间分辨率 (25μm) 的 PFOA 检测。观察到 PFOA 在斑马鱼肝脏组织中蓄积。H&E 染色结果证实了 PFOA 蓄积造成的损伤,与 MALDI MSI 结果一致。在暴露于 PFOA 的斑马鱼组中观察到 15 种磷脂种类的显著上调,这些磷脂在同一组织内表现出不同的空间分布。此外,在相同的组织切片中对不同的磷脂和 PFOA 进行共定位成像表明,PFOA 和磷脂之间可能存在两种不同的潜在相互作用,这需要进一步研究。MALDI-TOF-IMS 为探索新兴环境污染物的健康风险评估和生态毒理学中的内源性代谢物的原位空间分布和变化提供了一种新工具。

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