Relationship between hepatic and mitochondrial ceramides: a novel in vivo method to track ceramide synthesis.
Relationship between hepatic and mitochondrial ceramides: a novel in vivo method to track ceramide synthesis.
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DOI：
10.1016/j.jlr.2023.100366
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发表时间：
2023-05
影响因子：
6.5
通讯作者：
Parks, Elizabeth J
中科院分区：
文献类型：
--
作者：
Mucinski, Justine M;McCaffrey, Jonas M;Rector, R Scott;Kasumov, Takhar;Parks, Elizabeth J
神经酰胺（CERs）是参与线粒体功能障碍和多种代谢疾病发展的关键中间鞘脂。尽管越来越多的证据表明CER在疾病风险中的作用，但缺乏测量CER转换的动力学方法，特别是使用体内模型。通过口服溶解于饮用水中的13C3, 15N l-丝氨酸，定量测定10周龄C57Bl/6雄性和雌性小鼠的CER 18:1/16:0合成。为了生成同位素标记曲线，动物分别食用对照饮食或高脂肪饮食（HFD, n = 24只/日粮）2周，并在食用丝氨酸标记水的持续时间（0、1、2、4、7或12天，n = 4只动物/天/日粮）上变化。使用液相色谱串联质谱法对未标记和标记的肝脏和线粒体CER进行定量分析，两种饮食组之间的肝脏总CER含量没有差异，而线粒体总CER随着饲喂高脂肪饲料而增加（60%,P < 0.001）。在肝脏和线粒体池中，HFD诱导更高的饱和CER浓度（P < 0.05），并显著提高16:0线粒体CER的绝对周转率（线粒体：59%，P < 0.001，肝脏：15%，P = 0.256）。数据表明，由于HFD， cer的细胞重分布。这些数据表明，2周的HFD改变了线粒体cer的周转和含量。鉴于越来越多的数据表明CER有助于肝线粒体功能障碍和多种代谢性疾病的进展，这种方法现在可以用于研究在这些情况下CER的转换是如何改变的。
Ceramides (CERs) are key intermediate sphingolipids implicated in contributing to mitochondrial dysfunction and the development of multiple metabolic conditions. Despite the growing evidence of CER role in disease risk, kinetic methods to measure CER turnover are lacking, particularly using in vivo models. The utility of orally administered 13C3, 15N l-serine, dissolved in drinking water, was tested to quantify CER 18:1/16:0 synthesis in 10-week-old male and female C57Bl/6 mice. To generate isotopic labeling curves, animals consumed either a control diet or high-fat diet (HFD; n = 24/diet) for 2 weeks and varied in the duration of the consumption of serine-labeled water (0, 1, 2, 4, 7, or 12 days; n = 4 animals/day/diet). Unlabeled and labeled hepatic and mitochondrial CERs were quantified using liquid chromatography tandem MS. Total hepatic CER content did not differ between the two diet groups, whereas total mitochondrial CERs increased with HFD feeding (60%, P < 0.001). Within hepatic and mitochondrial pools, HFD induced greater saturated CER concentrations (P < 0.05) and significantly elevated absolute turnover of 16:0 mitochondrial CER (mitochondria: 59%, P < 0.001 vs. liver: 15%, P = 0.256). The data suggest cellular redistribution of CERs because of the HFD. These data demonstrate that a 2-week HFD alters the turnover and content of mitochondrial CERs. Given the growing data on CERs contributing to hepatic mitochondrial dysfunction and the progression of multiple metabolic diseases, this method may now be used to investigate how CER turnover is altered in these conditions.