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采用薄层层析、气相色谱和质谱联用技术研究 3T3-L1 脂肪细胞成熟过程中的脂质图谱差异。

Differences in the lipid patterns during maturation of 3T3-L1 adipocytes investigated by thin-layer chromatography, gas chromatography, and mass spectrometric approaches.

机构信息

Institute for Medical Physics and Biophysics, Medical Faculty, Leipzig University, Härtelstr. 16-18, 04107, Leipzig, Germany.

Leibniz Institute for Farm Animal Biology, Institute of Muscle Biology and Growth, Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany.

出版信息

Anal Bioanal Chem. 2020 Apr;412(10):2237-2249. doi: 10.1007/s00216-019-02243-w. Epub 2019 Dec 3.

DOI:10.1007/s00216-019-02243-w
PMID:31797017
Abstract

Populations of industrialized countries have registered a dramatically increasing prevalence in obesity for many years. Despite continuous research, mechanisms involved in the storage and utilization of chemical energy in adipocytes are still under investigation. Adipocytes have the task to store excessive energy in the form of triacylglycerols (TG) and it is already well-known that the fatty acyl composition of TG is largely determined by the composition of the diet. In contrast to TG, the composition of adipocyte phospholipids was less comprehensively investigated. In this study, the compositions of the most abundant phospholipid classes of 3T3-L1 undifferentiated (preadipocytes) and differentiated cells (adipocytes) were determined. The lipid fractions were isolated by normal phase high-performance thin-layer chromatography and subsequently analyzed by electrospray ionization mass spectrometry. Additionally, the fatty acyl (FA) compositions were determined by gas chromatography. The positions of the FA residues were additionally confirmed by phospholipase A digestion. The advantages and disadvantages of the different analytical approaches will be discussed. It will be shown that undifferentiated 3T3-L1 and mature adipocytes differ extremely regarding their compositions. This goes along with an increase in odd-chain fatty acids. Graphical abstract.

摘要

多年来,工业化国家的人口肥胖患病率显著增加。尽管进行了持续的研究,但脂肪细胞中化学能储存和利用的机制仍在研究中。脂肪细胞的任务是将多余的能量以三酰基甘油 (TG) 的形式储存,并且已经众所周知,TG 的脂肪酸组成在很大程度上取决于饮食的组成。与 TG 相比,脂肪细胞磷脂的组成研究得不够全面。在这项研究中,确定了 3T3-L1 未分化(前脂肪细胞)和分化细胞(脂肪细胞)中最丰富的磷脂类别的组成。通过正相高效薄层层析分离脂质馏分,然后通过电喷雾电离质谱进行分析。此外,通过气相色谱法测定脂肪酸 (FA) 组成。通过磷脂酶 A 消化进一步确认 FA 残基的位置。将讨论不同分析方法的优缺点。结果表明,未分化的 3T3-L1 和成熟的脂肪细胞在组成上差异极大。这伴随着奇数链脂肪酸的增加。

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