School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, China.
Zhangzhou Pien Tze Huang Pharmaceutical Co., Ltd., Zhangzhou, 363000, China.
Anal Chim Acta. 2022 Jun 15;1212:339885. doi: 10.1016/j.aca.2022.339885. Epub 2022 May 10.
Bile acid (BA) pool homeostasis can be disturbed when cholestasis, and ursodeoxycholic acid (UDCA) treatment is able to rectify the perturbed pool. Efforts were made here to propose a strategy enabling BAs-focused widely quantitative metabolomics and subsequently to lucubrate BA pool fluctuation trajectory in rats after dosing UDCA. A so-called universal metabolome standard (UMS) sample containing numerous natural BAs was constructed by involving various available BAs-enriched matrices. In-depth chemical characterization was conducted for UMS to capture as many BAs as possible. Diverse survey experiments were performed on Qtrap-MS to search BAs, and IT-TOF-MS was deployed to provide high-resolution m/z values for both precursor and fragment ion species. After structural annotation for 120 BAs, in total, online energy-resolved MS was subsequently programmed to pursue superior parameters for certain BAs, particularly BAs bearing two conjugated moieties. UMS was serially diluted to generate calibration sample set, and the regressive calibration curves, 120 ones in total, were responsible for converting each analyte response to quasi-content (1/dilution fold). The validated widely quasi-quantitative program was then applied to profile BA pool shift trajectories against time in UDCA- and vehicle-treated rats. Mild variations were observed for the quantitative pattern of BA pool from vehicle group, whereas UDCA could significantly shape BAs sub-metabolome. Significant increments occurred for the contents of not only the downstream BAs, but some upstream molecules in BA metabolic network. The shift levels of BAs were primarily governed by their distances away from UDCA. Fourteen differential BAs were involved for absolute quantitation to generate the concentration vs. time patterns, and dramatic upgradation occurred for most BAs, consolidating the variations transmitted from large-scale quasi-quantitative metabolomics. Above all, current study revealed the quasi-quantitative details of BA pool fluctuation initiated by UDCA-treatment, and more importantly, provided a promising approach for temporal BAs-targeted metabolomics.
胆汁酸 (BA) 池的动态平衡可能在胆汁淤积时受到干扰，熊去氧胆酸 (UDCA) 的治疗能够纠正失调的胆汁酸池。本研究旨在提出一种策略，使 BA 为中心的广泛定量代谢组学成为可能，并随后深入研究 UDCA 给药后大鼠胆汁酸池的波动轨迹。通过涉及各种可用的富含胆汁酸的基质，构建了一个所谓的通用代谢组标准 (UMS) 样本，其中包含了许多天然胆汁酸。对 UMS 进行了深入的化学表征，以尽可能多地捕获胆汁酸。在 Qtrap-MS 上进行了多种调查实验来搜索胆汁酸，并部署了 IT-TOF-MS 为前体和碎片离子种类提供高分辨率 m/z 值。对 120 种胆汁酸进行结构注释后，总共对在线能量分辨 MS 进行了编程，以寻求某些胆汁酸（特别是具有两个共轭部分的胆汁酸）的更好参数。UMS 被连续稀释以生成校准样本集，总共 120 个回归校准曲线负责将每个分析物的响应转换为准含量（1/稀释倍数）。经过验证的广泛准定量程序随后用于描绘 UDCA 和载体处理的大鼠中胆汁酸池随时间的变化轨迹。与载体组相比，胆汁酸池的定量模式观察到轻微变化，而 UDCA 可以显著塑造胆汁酸亚代谢组。不仅下游胆汁酸的含量增加，而且胆汁酸代谢网络中的一些上游分子的含量也增加。胆汁酸的转移水平主要由它们与 UDCA 的距离决定。有 14 种差异胆汁酸参与绝对定量以生成浓度随时间的变化模式，并且大多数胆汁酸发生了显著升级，这巩固了从大规模准定量代谢组学传递的变化。总之，本研究揭示了 UDCA 治疗引发的胆汁酸池波动的准定量细节，更重要的是，为时间相关的胆汁酸靶向代谢组学提供了一种有前途的方法。
