Analysis of Lipid Contents in Human Trabecular Meshwork Cells by Multiple Reaction Monitoring (MRM) Profiling Lipidomics.
Analysis of Lipid Contents in Human Trabecular Meshwork Cells by Multiple Reaction Monitoring (MRM) Profiling Lipidomics.
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DOI：
10.1007/978-1-0716-2966-6_24
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发表时间：
2023
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脂质是细胞的主要成分之一，发挥着许多重要的细胞功能。小梁网的脂质水平在维持房水引流和眼内压（IOP）稳态中起着重要作用。因此，表征房水流出通道组织中脂质含量的变化，以更好地了解其在维持IOP中的功能意义是很重要的。基于多反应监测（MRM）的分析有助于分析代谢组作为功能群的集合，并被用作探索性代谢组学和脂质组学方法。基于核磁共振成像的分析利用串联质谱实验在商用三重四极质谱仪上进行，该质谱仪具有三个对齐的四极质谱过滤器（Q1， q2和Q3）。该筛选方法可用于靶向脂质组学筛选。本章重点介绍了TM细胞的分离和培养方法，脂质提取，以及基于mrm的脂质组学方法和数据分析。
Lipids are among the major constituents of cells and play many important cellular functions. Lipid levels in the trabecular meshwork plays an important role in the maintenance of aqueous humor drainage and intraocular pressure (IOP) homeostasis. Therefore, it is important to characterize the changes in the lipid contents in the aqueous humor outflow pathway tissues to better understand their functional significance in the maintenance of IOP. The multiple reaction monitoring (MRM)-based profiling aids in the analysis of metabolome as a collection of functional groups and is utilized as an exploratory metabolomics and lipidomics approach. The MRM-based profiling utilizes tandem mass spectrometry experiments carried out on a commercial triple quadrupole mass spectrometer with three aligned quadrupole mass filters (Q1, q2, and Q3). This screening methodology can be utilized for targeted lipidomics screen. This chapter focuses on the methodology for isolation and culturing of the TM cells, lipid extraction, and MRM-based lipidomics approach with data analysis.