Identification and specific makers of abnormal bile acids excreted in inborn errors of bile acid biosynthesis

先天性胆汁酸生物合成缺陷中异常胆汁酸排泄的鉴定和特异性标志物

• 批准号: 21550091
• 负责人: IIDA Takashi
• 金额: $ 3万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21550091/
• 关键词: 生体分子; 異常胆汁酸代謝産物; LC/ESI-MS/MS; 先天性胆汁酸代謝異常症; Δ^4-3-オキソ-ステロイド-5β-レダクターゼ欠損症; 二次異常代謝産物; ステロイド多重抱合体; LC-MS; 3β-HSD/HIS欠損症; ニーマン-ピック病C型; LC/MS; NPC_1; LC; MS

The synthesis of bile salts from cholesterol is a complex biochemical pathway involving at least 16 enzymes. Most inborn errors of bile acid biosynthesis result in excessive formation of intermediates and/or their metabolites that accumulate in blood and are excreted in part in urine. Early detection is important as oral therapy with bile acids results in improvement. In the past, these intermediates in bile acid biosynthesis have been detected in neonatal blood or urine by screening with FAB-MS followed by detailed characterization using GC-MS. Both methods have proved difficult to automate, and currently most laboratories screen candidate samples using LC-MS/MS. Here, we describe a new, simple and sensitive analytical method for the identification and characterization of 39 conjugated and unconjugated bile acids, including Δ^-3-oxo- and Δ^<4, 6>-3-oxo-bile acids(markers for Δ^4-3-oxo-steroid 5β-reductase deficiency), using liquid chromatography-electrospray ionization tandem mass spectrometry(LC/ESI-MS/MS). In this procedure, a concentrated, desalted urinary sample(diluted with ethanol) is injected directly into the LC-ESI-MS/MS, operated with ESI and in the negative ion mode ; quantification is obtained by selected reaction monitoring(SRM). To evaluate the performance of our new method, we compared it to a validated method using GC-MS, in the analysis of urine from two patients with genetically confirmed Δ^4-3-oxo-steroid-5β-reductase deficiency as well as a third patient with an elevated concentration of abnormal conjugated and unconjugated Δ^4-3-oxo-bile acids. The Δ^4-3-oxo-bile acids concentration recovered in three patients with 5β-reductase deficiency were 48.8, 58.9, and 49.4μmol/mmol Creatinine, respectively by LC/ESI-MS/MS.

胆固醇中胆汁盐的合成是一种涉及至少16个酶的复杂生化途径。胆汁酸生物合成的大多数天生误差会导致中间体和/或其代谢产物的过量形成，这些中间体积聚在血液中，部分在尿液中非常极端。早期检测很重要，因为口服治疗和胆汁酸会改善。过去，通过使用Fab-MS筛选，然后使用GC-MS进行详细表征，在新生儿血液或尿液中检测到了这些中间体。事实证明，两种方法都难以自动化，目前大多数实验室使用LC-MS/MS筛选了候选样品。在这里，我们描述了一种新的，简单且敏感的分析方法，用于鉴定和表征39种共轭和未偶联的胆汁酸，包括δ^^^^^^^-3-氧和δ^<4，6> -3-氧气胆酸（使用Δ^4-3-3-OXO-s-oxo-seroid-seroid-seroid s-Rectose s riqudy Chromation Chromatormation-chromation-chromemation-chromecraight）光谱法（LC/ESI-MS/MS）。在此过程中，将浓缩的脱盐尿液样品（用乙醇稀释）直接注入LC-ESI-MS/MS，并以ESI和负离子模式运行；通过选定的反应监测（SRM）获得定量。为了评估我们的新方法的性能，我们将其与使用GC-MS进行了经过验证的方法进行了比较，在两名患者的尿液分析中，有两名普遍确认的Δ^4-3-氧气 - 螺旋-5β-还原酶缺乏症以及第三名患者，患者浓度升高，异常偶联且未缀合的δ^^4-3-氧化二氧化物。 LC/ESI-MS/MS分别在三名5β-还原酶缺乏症患者中回收的δ^4-3-氧胆酸盐浓度分别为48.8、58.9和49.4μmol/mmol肌酐。

项目成果

新生児の胆嚢および腸管内胎便中の胆汁酸分析

新生儿胆囊和肠胎粪中的胆汁酸分析

• 发表时间: 2011
• 作者: 武藤晃奈;武井一;吽野篤;入戸野博;佐藤洋明;村井毅;黒澤隆夫;藤間貞彦;飯田隆
• 通讯作者: 飯田隆

Relative Inhibitory Activity of Bile Acids Against 12-O-Tetradecanolyphorbol-13-acetate-Induced Inflammation, and Chenodeoxycholic Acid Inhibition of Tumour Promotion in Mouse Skin Two-Stage Carcinogenesis

胆汁酸对 12-O-Tetradecanolyphorbol-13-acetate 诱导的炎症的相对抑制活性，以及​​鹅去氧胆酸对小鼠皮肤两阶段癌变中肿瘤促进的抑制作用

• 发表时间: 2009
• 期刊: J. Pharm. Pharmacol
• 作者: K. Yasukawa;T. Iida;Y. Fujimoto
• 通讯作者: Y. Fujimoto

Chemical synthesis of the (25R)-and (25S)-epimers of 3αa,7α,12α-trihydroxy-5α-cholestan-27-oic acid as well as their corresponding glycine and taurine conjugates

3αa,7α,12α-三羟基-5α-胆甾坦-27-oic 酸的 (25R)-和 (25S)-差向异构体及其相应的甘氨酸和牛磺酸缀合物的化学合成

• 发表时间: 2011
• 期刊: Chemistry & Physics of Lipids
• 影响因子: 3.4
• 作者: S.Ogawa;K.Mitamura;S.Ikegawa;M.D.Krasowski;L.R.Hagey;A.F.Hofmann;T.Iida
• 通讯作者: T.Iida

Characterization of Non-Enzymatic Acylation of Amino or Thiol Groups of Binucleophiles by the Acyl-Adenylate or Acyl-CoA Thioester of Cholic Acid

胆酸的酰基腺苷酸或酰基辅酶A硫酯对双亲核试剂的氨基或硫醇基团的非酶酰化的表征

• 发表时间: 2011
• 期刊: Anal. Bioanal. Chem
• 作者: K. Mitamura;E. Aoyama;T. Sakai;T. Iida;A. F. Hofmann;S. Ikegawa
• 通讯作者: S. Ikegawa

Monoterpene Glucosides from Ziziphora clinopodioides(Labiatae)

来自 Ziziphora clinopodioides（唇形科）的单萜糖苷

• 发表时间: 2012
• 期刊: Chem. Pharm. Bull
• 作者: Megumi Furukawa;Naoki Oikawa;Toru Imohata;Mitsuko Makino;Shoujiro Ogawa;Takashi Iida;Yasuo Fujimoto and Susumu Kitanaka
• 通讯作者: Yasuo Fujimoto and Susumu Kitanaka