Precolumn derivatization LC/MS method for observation of efficient hydrogen sulfide supply to the kidney via d-cysteine degradation pathway
Precolumn derivatization LC/MS method for observation of efficient hydrogen sulfide supply to the kidney via d-cysteine degradation pathway
复制标题
柱前衍生 LC/MS 方法观察通过 d-半胱氨酸降解途径向肾脏有效供应硫化氢
DOI：
10.1016/j.jpba.2022.115088
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发表时间：
2023
影响因子：
3.4
通讯作者：
Todoroki Kenichiro
中科院分区：
文献类型：
--
作者：
Sugiyama Eiji;Higashi Toru;Nakamura Mao;Mizuno Hajime;Toyo’oka Toshimasa;Todoroki Kenichiro
D-半胱氨酸(d-Cys)通过D-氨基酸氧化酶(DAO)/3-硫代丙酮酸硫转移酶途径代谢成硫化氢。该途径是补充硫化氢以改善小鼠口服d-半胱氨酸后的急性肾损伤所必需的。然而，DAO的限速活性是否调节组织选择性或d-半胱氨酸降解和H_2S补充的程度仍不清楚。在这里，为了分析d-Cys和H_2S的含量，我们使用了两种衍生化方法，一种是检测不到半胱氨酸异构化的新方法，另一种是建立的H_2S的分析方法。采用LC/MS法，色谱柱为C18柱。结果表明，抑制DAO可显著抑制小鼠肾脏对H_2S的补充和d-半胱氨酸的降解。此外，我们还发现d-半胱氨酸在肾脏中比l-半胱氨酸更有效地代谢成H_2S。我们的结果揭示了该方法的实用性，并支持了d-Cys给药在改善肾脏的H_2S供应方面的优势。
d-Cysteine (d-Cys) is metabolized to hydrogen sulfide (H2S) byd-amino acid oxidase (DAO)/3-mercaptopyruvate sulfurtransferase pathway. The pathway is required for H2S supplementation that ameliorates acute kidney injury after the oral administration ofd-Cys in mice. However, whether the rate-limiting activity of DAO regulates the tissue-selectivity or the extent ofd-Cys degradation and H2S supplementation remains unclear. Here, to analyze the levels ofd-Cys and H2S, we use two derivatization methods, a new method with no detectable isomerization of Cys and an established method for H2S. The derivatives were determined by LC/MS using a C18 column. With the methods, we show that inhibition of DAO significantly suppresses the H2S supplementation andd-Cys degradation in the mouse kidney. Additionally, we found thatd-Cys is more efficiently metabolized into H2S thanl-Cys in the kidney. Our results reveal the utility of the method and support the advantage ofd-Cys administration in improving the supply of H2S to the kidneys.