Anti‐inflammatory activities and glycerophospholipids metabolism in KLA‐stimulated RAW 264.7 macrophage cells by diarylheptanoids from the rhizomes of Alpinia officinarum

Alpinia officinarum is used for its anti‐inflammatory activity historically in China. Diarylheptanoids isolated from A. officinarum play important biological roles in the prevention and treatment of inflammatory disorders. Seven diarylheptanoids (1–7) were isolated from A. officinarum. The cell viabilities and anti‐inflammatory activities of diarylheptanoids were evaluated by MTT assay and tumor necrosis factor‐α production in Kdo2‐lipid A‐stimulated RAW 264.7 cells in vitro. The relationships between their anti‐inflammatories and structure‐activities are discussed. The results indicated that compounds 1 and 3–7 had significant anti‐inflammatory activities. The relationships between inflammation and phospholipids metabolism were elucidated by multivariate data analysis. Twenty‐two potential biomarkers were identified in inflammatory group vs. blank group, and 11 potential biomarkers were identified for inflammatory group vs. drug‐treatment groups. Ten common phospholipids were characterized. On the basis of a previous study in our laboratory, we found that phosphatidylethanolamine (18:0/18:1) might be the important glycerophospholipid biomarker in inflammation. In this study, we firstly combined anti‐inflammatory activities and glycerophospholipids changes of traditional Chinese medicine. This work suggests that the anti‐inflammatory activities of diarylheptanoids might be significantly related to glycerophospholipids and could provide a useful database for investigating the anti‐inflammatory effects of traditional Chinese medicine.     

基于衍生化技术的甘油磷脂分析方法研究进展

磷脂是所有生物细胞膜的主要成分,在许多生命活动过程中具有重要的功能。但由于生物样本中的磷脂种类繁多,含量极低,存在基质抑制效应,且结构中缺少易电离的官能团,从而导致对磷脂的定性和定量分析较困难。利用化学衍生化技术对其进行结构修饰可以提高离子化效率、改善色谱分离度且提高质谱(MS)检测的灵敏度和选择性。MS与衍生化方法结合已被广泛用于蛋白组学、糖组学、代谢物等的分析。近年来,这一策略逐渐被应用于脂质组学的分析研究。该文综述了国内外近10年基于衍生化技术的甘油磷脂分析方法及其应用研究进展,以激发衍生化技术在脂质组学分析中的应用潜能。     

Rapid identification of glycerophospholipids from RAW264.7 cells by UPLC/ESI ‐QTOF‐MS

The study aims to develop a rapid, sensitive ultra‐performance liquid chromatography coupled with an electrospray ionization quadruple time‐of‐flight tandem mass spectrometry (UPLC/Q‐TOF‐MS) analytical method for identifying glycerophospholipids (GPLs) from RAW264.7 cells. A total of 78 GPLs including 22 phosphatidylethanolamines (PEs), 49 phosphatidylcholines (PCs), four phosphatidylglycerols, one phosphatidylinositol and two unknown GPLs were identified. PC (14:0/16:1), PC (14:0/16:0), PE (0:0/20:3), PE (22:5/0:0) and PE (22:3/0:0) were identified for the first time. The UPLC/Q‐TOF‐MS method is suitable for targeting analysis of GPLs from RAW264.7 cells, which allows us to find out new GPLs compositions related to inflammatory diseases and to explain their pharmacological roles in inflammatory process. Copyright © 2014 John Wiley & Sons, Ltd.     

Isotopomer analysis of lipid biosynthesis by high resolution mass spectrometry and NMR

We have coupled 2D-NMR and infusion FT-ICR-MS with computer-assisted assignment to profile ¹³C-isotopologues of glycerophospholipids (GPL) directly in crude cell extracts, resulting in very high information throughput of >3000 isobaric molecules in a few minutes. A mass accuracy of better than 1 ppm combined with a resolution of 100,000 at the measured m/z was required to distinguish isotopomers from other GPL structures. Isotopologue analysis of GPLs extracted from LCC2 breast cancer cells grown on [U-¹³C]-glucose provided a rich trove of information about the biosynthesis and turnover of the GPLs. The isotopologue intensity ratios from the FT-ICR-MS were accurate to ≈1% or better based on natural abundance background, and depended on the signal-to-nose ratio. The time course of incorporation of ¹³C from [U-¹³C]-glucose into a particular phosphatidylcholine was analyzed in detail, to provide a quantitative measure of the sizes of glycerol, acetyl CoA and total GPL pools in growing LCC2 cells. Independent and complementary analysis of the positional ¹³C enrichment in the glycerol and fatty acyl chains obtained from high resolution 2D NMR was used to verify key aspects of the model. This technology enables simple and rapid sample preparation, has rapid analysis, and is generally applicable to unfractionated GPLs of almost any head group, and to mixtures of other classes of metabolites.     

Lipid analysis by thin-layer chromatography--a review of the current state

High-performance thin-layer chromatography (HPTLC) is a widely used, fast and relatively inexpensive method of separating complex mixtures. It is particularly useful for smaller, apolar compounds and offers some advantages over HPLC. This review gives an overview about the special features as well as the problems that have to be considered upon the HPTLC analysis of lipids. The term "lipids" is used here in a broad sense and comprises fatty acids and their derivatives as well as substances related biosynthetically or functionally to these compounds. After a short introduction regarding the stationary phases and the methods how lipids can be visualized on an HPTLC plate, the individual lipid classes will be discussed and the most suitable solvent systems for their separation indicated. The focus will be on lipids that are most abundant in biological systems, i.e. cholesterol and its derivates, glycerides, sphingo- and glycolipids as well as phospholipids. Finally, a nowadays very important topic, the combination between HPTLC and mass spectrometric (MS) detection methods will be discussed. It will be shown that this is a very powerful method to investigate the identities of the HPTLC spots in more detail than by the use of common staining methods. Future aspects of HPTLC in the lipid field will be also discussed.     

N3-羟烷基替加氟制备方法改良及其环甘油磷脂缀合物的合成与抗肿瘤活性

在药物-载体复合型前药的研究中,磷脂-药物缀合物(如脂核苷酸)特别引人注目,它以磷脂作为载体,与核苷等药物分子以共价连接形成的一类新型脂核苷酸前药[1～3].     

腺苷环甘油磷脂缀合物的合成及性质研究

以碘活化的六乙基亚磷酰三胺为磷酰化及环化试剂，经一锅法合成并分离得到硫代环磷酸酯的两个非对映异构体，并对这两个非对映异构体的性质进行了研究．