Comprehensive and sensitive quantification of long-chain and very long-chain polyunsaturated fatty acids in small samples of human and mouse retina
Comprehensive and sensitive quantification of long-chain and very long-chain polyunsaturated fatty acids in small samples of human and mouse retina
复制标题
DOI：
10.1016/j.chroma.2013.07.103
复制
发表时间：
2013-09-13
影响因子：
4.1
通讯作者：
Bernstein, Paul S.
中科院分区：
文献类型：
--
作者：
Liu, Aihua;Terry, Ryan;Bernstein, Paul S.
脂肪酸 (FA)，包括长链和超长链多不饱和脂肪酸（LC-PUFA、C12-22 VLC-PUFA、C24-38），在视网膜功能和健康中发挥着重要作用。 LC-PUFA 和 VLC-PUFA 的缺乏以及负责超长链脂肪酸延长的酶 (ELOVL4) 的突变与黄斑营养不良和退化有关。已发表的分析方法，包括高效液相色谱-质谱联用 (HPLC-MS) 和气相色谱-MS (GC-MS)，可以量化 VLC-PUFA，但至少需要整个人类视网膜，这限制了了解视网膜内区域水平可能发生的脂质生理相关变化的能力。到目前为止，由于人类黄斑尺寸较小（直径 4-5 毫米），因此对 VLC-PUFA 进行定量尚不可行。黄斑是视网膜中央富含视锥细胞的区域，负责高敏锐度视力。在这项研究中，我们开发了一种灵敏的 GC-MS 方法，使用新一代增强 GC-MS 检测器灵敏度，首次不仅量化了 14 个 VLC-PUFA 和 26 个 LC-FA，还量化了人类视网膜 4 毫米冲孔或一对小鼠视网膜中 PUFA 的 n-3/n-6 比率。我们的结果表明，人类周边视网膜中的饱和 LC-FA 含量高于黄斑中的含量，而不饱和的 LC-FA 含量在黄斑中的含量高于周边视网膜中的含量。另一方面，与黄斑相比，周边视网膜中的 VLC-PUFA 含量更高。在人类黄斑和周边视网膜之间观察到的 PUFA 的 n-3/n-6 比率没有差异，而小鼠视网膜的 VLC-PUFA 几乎是人类黄斑和周边视网膜的十倍（分别为 2.27% 对 0.25% 和 0.32%），并且与人类视网膜相比更高的 n-3/n-6 比率（9:1 对相似的 0.9:1）。这种高灵敏度分析技术为研究 FA 在黄斑变性和营养不良的病理过程中的作用提供了一个有价值的新工具。 (C) 2013 Elsevier B.V. 保留所有权利。
Fatty acids (FAs), including long-chain and very long-chain polyunsaturated fatty acids (LC-PUFAs, C12-22 VLC-PUFAs, C24-38), play an important role in retinal function and health. Deficiencies in LC-PUFAs and VLC-PUFAs, as well as mutations in the enzyme responsible for elongation of very long-chain fatty acids (ELOVL4), have been associated with macular dystrophies and degenerations. Published analytical methods, including high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) and gas chromatography-MS (GC-MS), can quantify VLC-PUFAs but require at least an entire human retina which limits the ability to understand physiologically relevant variations in lipids that can occur at a regional level within the retina. Until now, quantification of VLC-PUFAs in just the human macula, the cone-rich region of the central retina responsible for high acuity vision, has not been feasible due to its small size (4-5 mm in diameter). In this study, we have developed a sensitive GC-MS method using newer generation enhanced GC-MS detector sensitivity which for the first time quantifies not only 14 VLC-PUFAs and 26 LC-FAs but also n-3/n-6 ratios of PUFAs in 4 mm punches of human retina or a single pair of mouse retinas. Our results showed that saturated LC-FAs are higher in the human peripheral retina than in the macula, while unsaturated LC-FAs are higher in the macula than in the peripheral retina. On the other hand, the VLC-PUFAs are higher in the peripheral retina compared to macula. There is no difference in n-3/n-6 ratios of PUFAs observed between human macula and peripheral retina, while mouse retina has almost ten times more VLC-PUFAs than human macula and peripheral retina (2.27% versus 0.25% and 0.32%, respectively) and much higher n-3/n-6 ratios compared to human retina (9:1 versus similar to 0.9:1). This high sensitivity analytical technique provides a valuable new tool for studies on the role of FAs in the pathological processes of macular degenerations and dystrophies. (C) 2013 Elsevier B.V. All rights reserved.