Identification of neutral and acidic glycosphingolipids in the human dermal fibroblasts
Identification of neutral and acidic glycosphingolipids in the human dermal fibroblasts
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DOI：
10.1016/j.ab.2019.113348
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发表时间：
2019-09-15
影响因子：
2.9
通讯作者：
Cataldi, Tommaso R. I.
中科院分区：
文献类型：
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作者：
Calvano, Cosima Damiana;Ventura, Giovanni;Cataldi, Tommaso R. I.
皮肤成纤维细胞被认为是能够反映时间和生物老化的原代人类细胞的有价值的模型。在这里，脂质组学研究鞘糖脂（GSL）发生在容易接近的人皮肤成纤维细胞（HDF）。采用负电喷雾离子化反相液相色谱法（RPLC-ESI）结合轨道阱或线性离子阱多级质谱法，对商业成人和新生儿原代人成纤维细胞以及成人志愿者皮肤样本中的GSL进行表征。在负离子模式下的碰撞诱导解离使我们能够获得有关单糖数和神经酰胺组成的信息，而神经酰胺阴离子的串联质谱可用于识别鞘氨醇碱。成功识别了近60种内源性GSL物质，即33种己糖基神经酰胺（即，HexCer、Hex（2）Cer和Hex（3）Cer）和24种神经节苷脂（单唾液酸GM 1、GM 2和GM 3）以及沿着5种球状苷脂Gb 4。获得了GSL的平均含量，皮肤成纤维细胞中最具代表性的GSL是Hex（3）Cer，也称为Gb（3）Cer，其次是Gb 4、HexCer和Hex（2）Cer，而神经节苷脂几乎不可定量。在检查的细胞系中最丰富的GSL共享相同的神经酰胺碱（Le. d18：1），相对含量为d18：1/24：1 > d18：1/24：0 > d18：1/16：0 > d18：1/22：0。
Skin fibroblasts are recognized as a valuable model of primary human cells able of mirroring the chronological and biological aging. Here, a lipidomic study of glycosphingolipids (GSL) occurring in the easily accessible human dermal fibroblasts (HDF) is presented. Reversed-phase liquid chromatography with negative electrospray ionization (RPLC-ESI) coupled to either orbitrap or linear ion-trap multiple-stage mass spectrometry was applied to characterize GSL in commercially adult and neonatal primary human fibroblast cells and in skin samples taken from an adult volunteer. Collision-induced dissociation in negative ion mode allowed us to get information on the monosaccharide number and ceramide composition, whereas tandem mass spectra on the ceramide anion was useful to identify the sphingoid base. Nearly sixty endogenous GSL species were successfully recognized, namely 33 hexosyl-ceramides (i.e., HexCer, Hex(2)Cer and Hex(3)Cer) and 24 gangliosides as monosialic acid GM1, GM2 and GM3, along with 5 globosides Gb4. An average content of GSLs was attained and the most representative GSL in skin fibroblasts were Hex(3)Cer, also known as Gb(3)Cer, followed by Gb4, HexCer and Hex(2)Cer, while gangliosides were barely quantifiable. The most abundant GSLs in the examined cell lines share the same ceramide base (Le. d18:1) and the relative content was d18:1/24:1 > d18:1/24:0 > d18:1/16:0 > d18:1/22:0.