In situ metabolic profile and spatial distribution of ocular tissues: New insights into dry eye disease
In situ metabolic profile and spatial distribution of ocular tissues: New insights into dry eye disease
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DOI：
10.1016/j.jtos.2021.12.013
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发表时间：
2022-01-17
期刊：
影响因子：
6.4
通讯作者：
Wang, Liqiang
中科院分区：
文献类型：
--
作者：
Chen, Xiaoniao;Zhang, Chuyue;Wang, Liqiang
目的：干眼病（DED）是一种影响数百万人的慢性多因素疾病，其发病机制尚不清楚。基质辅助激光解吸电离质谱成像（MALDI-MSI）是一种结合高通量质谱和分子成像的新型原位可视化方法。我们的目的是通过MALDI-MSI研究眼内原位代谢变化，以加速对DED发病机制的认识。方法：采用东莨菪碱皮下注射建立实验性干眼模型。通过泪膜破裂时间、荧光素钠、组织病理学染色、细胞凋亡等指标评价其诱导程度。应用MALDI- msi对DED大鼠进行原位眼部代谢组学检测，并采用同切片的组织病理学染色与MALDI进行并排比较，以标注眼部不同的组织结构。结果：考虑到眼部组织的复杂性，我们可视化了特定眼部区域（角膜中央、角膜周围、穹窿结膜、眼睑结膜和房水）的代谢物，并鉴定出与DED相关的代谢物，具有相对丰度和空间特征信息。此外，综合通路分析表明，在干眼组织的某些区域，甘油磷脂、鞘脂、苯丙氨酸等代谢通路以及甘氨酸、丝氨酸和苏氨酸的代谢发生了显著改变。此外，我们还讨论了具有时空特征的代谢途径如何参与DED过程。结论：我们的数据利用MALDI-MSI原位分析的优势，准确分析了DED中特定区域的代谢行为，为揭示DED的发病机制提供了新的线索。
Purpose: Dry eye disease (DED) is a chronic multifactorial disorder affecting millions of people, yet the pathogenesis mechanisms still remain unclear. Matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) is a novel in situ visualization approach combined high-throughput mass spectrometry and molecular imaging. We aimed to explore the in situ ocular metabolic changes via MALDI-MSI to accelerate the recognition of DED pathogenesis.Methods: Experimental dry eye was established in Wistar rats by subcutaneous injection of scopolamine. The induction of DED was assessed by tear film breakup time, sodium fluorescein, histopathological staining and cell apoptosis. MALDI-MSI was applied to explore in situ ocular metabolomic in DED rats, and histopathological staining from same sections were used for side-by-side comparison with MALDI to annotate different tissue structures in the eye.Results: Considering the complexity of ocular tissue, we visualized the metabolites in specific ocular regions (central cornea, peripheral cornea, fornix conjunctiva, eyelid conjunctiva and aqueous humor), and identified metabolites related to DED, with information of relative abundance and spatial signatures. In addition, integrative pathway analysis illustrated that, several metabolic pathways such as glycerophospholipid, sphingolipid phenylalanine, and metabolism of glycine, serine and threonine were significantly altered in certain regions in the dry eye tissue. Moreover, we discussed how the metabolic pathways with spatiotemporal signatures might be involved in the DED process.Conclusions: Our data exploit the advantages of in situ analysis of MALDI-MSI to accurately analyze the region specific metabolic behaviors in DED, and provide new clues to uncover DED pathogenesis.