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用于诊断葡聚糖硫酸钠诱导的结肠炎模型中炎症性肠病动态变化的顶空-气相色谱-质谱联用方法

HS-GC-MS Method for the Diagnosis of IBD Dynamics in a Model of DSS-Induced Colitis.

作者信息

Shagaleeva Olga Yu, Kashatnikova Daria A, Kardonsky Dmitry A, Efimov Boris A, Ivanov Viktor A, Smirnova Svetlana V, Zorkina Yana A, Vorobjeva Elizaveta A, Silantiev Artemiy S, Kazakova Viktoriia D, Kolesnikova Irina V, Markelova Maria I, Vanyushkina Anna A, Chaplin Andrei V, Grigoryeva Tatiana V, Zakharzhevskaya Natalya B

机构信息

Laboratory of Molecular Pathophysiology, Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Moscow, Russia.

Laboratory of Ecological Genetics, Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia.

出版信息

Bio Protoc. 2025 Mar 20;15(6):e5246. doi: 10.21769/BioProtoc.5246.

DOI:10.21769/BioProtoc.5246
PMID:40395844
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12086341/
Abstract

Inflammatory bowel disease (IBD) is highly prevalent globally and, in the majority of cases, remains asymptomatic during its initial stages. The gastrointestinal microbiota secretes volatile organic compounds (VOCs), and their composition alters in IBD. The examination of VOCs could prove beneficial in complementing diagnostic techniques to facilitate the early identification of IBD risk. In this protocol, a model of sodium dextran sulfate (DSS)-induced colitis in rats was successfully implemented for the non-invasive metabolomic assessment of different stages of inflammation. Headspace-gas chromatography-mass spectrometry (HS-GC-MS) was used as a non-invasive method for inflammation assessment at early and remission stages. The disease activity index (DAI) and histological method were employed to assess intestinal inflammation. The HS-GC-MS method demonstrated high sensitivity to intestine inflammation, confirmed by DAI and histology assay, in the acute and remission stages, identifying changes in the relative content of VOCs in stools. HS-GC-MS may be a useful and non-invasive method for IBD diagnostics and therapy effectiveness control. Key features • Experiments performed in vivo to better control DSS-induced colon damage and to aid the study of IBD development in humans. • Optimized for the following organisms: Wistar rats and C57BL/6 mice. • An easily assessed disease activity index (DAI) (weight loss, stool consistency, degree of fecal occult blood) and histopathological examination are suggested for additional IBD confirmation. • Enables VOC testing with relatively small stool samples.

摘要

炎症性肠病(IBD)在全球范围内高度流行,且在大多数情况下,其初始阶段无症状。胃肠道微生物群会分泌挥发性有机化合物(VOCs),IBD患者体内这些化合物的组成会发生改变。检测VOCs可能有助于补充诊断技术,以促进IBD风险的早期识别。在本方案中,成功建立了硫酸葡聚糖钠(DSS)诱导的大鼠结肠炎模型,用于对炎症不同阶段进行非侵入性代谢组学评估。顶空气相色谱 - 质谱联用(HS - GC - MS)被用作在炎症早期和缓解期进行炎症评估的非侵入性方法。采用疾病活动指数(DAI)和组织学方法评估肠道炎症。HS - GC - MS方法对肠道炎症表现出高灵敏度,在急性和缓解期经DAI和组织学检测证实,可识别粪便中VOCs相对含量的变化。HS - GC - MS可能是一种用于IBD诊断和治疗效果监测的有用且非侵入性的方法。关键特性 • 在体内进行实验,以更好地控制DSS诱导的结肠损伤,并有助于研究人类IBD的发展。 • 针对以下生物体进行了优化:Wistar大鼠和C57BL / 6小鼠。 • 建议使用易于评估的疾病活动指数(DAI)(体重减轻、粪便稠度、粪便潜血程度)和组织病理学检查进行额外的IBD确认。 • 能够使用相对少量的粪便样本进行VOC检测。

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Bull Exp Biol Med. 2024 Sep;177(5):626-629. doi: 10.1007/s10517-024-06237-2. Epub 2024 Sep 30.
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