A two-step high-performance liquid chromatography (HPLC) method for the profiling and qualitative assessment of oxidized phospholipids (oxPL) present in foods was developed. The applicability of the investigated two-step HPLC protocol was verified for separation of enzymatically and thermally oxidized hen egg yolk phospholipids (PL) as a relevant food model. In the first step, seven individual PL classes were separated using hydrophilic interaction liquid chromatography (HILIC). The collected fractions of two main egg yolk PL classesphosphatidylethanolamines and phosphatidylcholineswere further directed to the second step of separation aimed at profiling and qualitative assessment of their oxidized species. For this purpose, the reverse phase (RP) chromatography coupled to charged aerosol, ultraviolet detection (UV) and mass spectrometry detection were employed. A database of potential oxPL including primary (hydroperoxides) and long-chain secondary PL oxidation products (epoxides, alcohols, and ketones) as well as some of their possible combinations was created. Additionally, the results were compared with the profiles of PL hydroperoxides obtained using thin layer chromatography (TLC) with N,N-dimethyl-p-phenylenediamine visualization.
Phospholipids (PLs) are important biomolecules that not only constitute structural building blocks and scaffolds of cell and organelle membranes but also play a vital role in cell biochemistry and physiology. Moreover, dietary exogenous PLs are characterised by high nutritional value and other beneficial health effects, which are confirmed by numerous epidemiological studies. For this reason, PLs are of high interest in lipidomics that targets both the analysis of membrane lipid distribution as well as correlates composition of lipids with their effects on functioning of cells, tissues and organs. Lipidomic assessments follow-up the changes occurring in living organisms, such as free radical attack and oxidative modifications of the polyunsaturated fatty acids (PUFAs) build in PL structures. Oxidised PLs (oxPLs) can be generated exogenously and supplied to organisms with processed food or formed endogenously as a result of oxidative stress. Cellular and tissue oxPLs can be a biomarker predictive of the development of numerous diseases such as atherosclerosis or neuroinflammation. Therefore, suitable high-throughput analytical techniques, which enable comprehensive analysis of PL molecules in terms of the structure of hydrophilic group, fatty acid (FA) composition and oxidative modifications of FAs, have been currently developed. This review addresses all aspects of PL analysis, including lipid isolation, chromatographic separation of PL classes and species, as well as their detection. The bioinformatic tools that enable handling of a large amount of data generated during lipidomic analysis are also discussed. In addition, imaging techniques such as confocal microscopy and mass spectrometry imaging for analysis of cellular lipid maps, including membrane PLs, are presented.
The blood–brain barrier (BBB) tightly controls the microenvironment of the central nervous system (CNS) to allow neurons to function properly. Additionally, emerging studies point to the beneficial effect of natural oils affecting a wide variety of physiological and pathological processes in the human body. In this study, using an in vitro model of the BBB, we tested the influence of natural fish oil mixture (FOM) vs. borage oil (BO), both rich in long-chain polyunsaturated fatty acids (LC-PUFAs) and monounsaturated fatty acids (MUFAs) such as oleic acid (C18:1n9c) or nervonic acid (NA), on human oligodendrocyte precursor cells (hOPCs) during their maturation to oligodendrocytes (OLs) regarding their ability to synthesize myelin peptides and NA. We demonstrated that FOM, opposite to BO, supplemented endothelial cells (ECs) and astrocytes forming the BBB, affecting the function of hOPCs during their maturation. This resulted in improved synthesis of myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), proteolipid protein (PLP), and NA in mature OLs. This effect is probably the result of BBB cell and hOPC stimulation via free fatty acid receptors (FFARs), which increases insulin growth factor-1 (IGF-1), ciliary neurotrophic factor (CNTF), and brain-derived neurotrophic factor (BDNF) and inhibits fibroblast growth factor 2 (FGF-2) synthesis. The unique formula of fish oil, characterized by much more varied components compared to those of BOs, also improved the enhancement of the tight junction by increasing the expression of claudin-5 and VE-cadherin on ECs. The obtained data justify consideration of naturally derived fish oil intake in human diet as affecting during remyelination.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.