New acylceramide in native and reconstructed epidermis
New acylceramide in native and reconstructed epidermis
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DOI：
10.1046/j.1523-1747.2003.12103.x
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发表时间：
2003-04-01
影响因子：
6.5
通讯作者：
Wertz, P
中科院分区：
文献类型：
--
作者：
Ponec, M;Weerheim, A;Wertz, P
在体外条件下，在气液界面培养正常人角质形成细胞可形成完全分化的表皮。虽然重建的表皮与天然组织非常相似，但在角质层脂质分布和细胞间脂质组织方面仍存在一些差异。由于神经酰胺属于角质层主要脂类之一，本研究的目的是更详细地表征这部分。为此，通过柱层析分离单个神经酰胺组分，并结合核磁共振波谱、高效薄层色谱和气相色谱进行表征。本研究结果表明，在天然和重建的人表皮中，可提取的神经酰胺部分除了含有已知的酰基神经酰胺（EOS， EOH）外，还含有一种新的酰基神经酰胺，其中ω - o -酰基羟基与植物鞘氨酸（EOP）酰胺相连。同样的三个鞘基（S， P， H）也存在于酰胺连接的非羟基和α -羟基酸的神经酰胺中。尽管两种组织中存在相同类型的神经酰胺，但在它们的脂肪酸谱中发现了一些差异。重建表皮中3种酰基神经酰胺组分中亚油酸含量均显著降低；神经酰胺（NS）部分富含短脂肪酸，神经酰胺（AS）部分富含长链α -羟基酸。这些差异加上较低的游离脂肪酸含量可以解释早期x射线衍射观察到的角质层脂质组织中天然组织和重建组织之间的差异。
Culturing of normal human keratinocytes at the air-liquid interface results in the formation of fully differentiated epidermis under in vitro conditions. Although the reconstructed epidermis shows a close resemblance to native tissue, there are still some differences in the stratum corneum lipid profile and intercellular lipid organization. As ceramides belong to one of the major stratum corneum lipid classes, the aim of this study was to characterize this fraction in more detail. For this purpose, individual ceramide fractions were isolated by column chromatography and characterized by a combination of nuclear magnetic resonance spectroscopy, high-performance thin-layer chromatography, and gas chromatography. The results of this study show that in both the native and reconstructed human epidermis the extractable ceramide fraction contains, in addition to the well known acylceramides (EOS, EOH), a new acylceramide in which the omega-O-acylhydroxyacid is amide-linked to phytosphingosine (EOP). The same three sphingoid base moieties (S, P, H) are also found in ceramides with amide-linked nonhydroxy and alpha-hydroxyacids. Whereas the same types of ceramides were present in both tissues, some differences in their fatty acid profiles have been found. In reconstructed epidermis the content of linoleic acid in all three acylceramides fraction was significantly, lower; the ceramide (NS) fraction was enriched in short fatty acids and the ceramide(AS) fraction was enriched in long chain alpha-hydroxyacids. These differences together with a lower content of free fatty acids may explain the differences between native and reconstructed tissue in stratum corneum lipid organization observed earlier by X-ray diffraction.