Effect of Shaoyaotang on Fecal Metabolites in Rats with Ulcerative Colitis Based on Liquid Chromatography-mass Spectrometry

LIU Xingci; WU Dongsheng; CAO Hui; LU Yi; LI Yanhong; YANG Yuting; XIE Nianjia

doi:10.13422/j.cnki.syfjx.20230201

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Effect of Shaoyaotang on Fecal Metabolites in Rats with Ulcerative Colitis Based on Liquid Chromatography-mass Spectrometry

更新时间：2023-03-30

- Effect of Shaoyaotang on Fecal Metabolites in Rats with Ulcerative Colitis Based on Liquid Chromatography-mass Spectrometry
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 9, Pages: 61-70(2023)
- 作者机构：
  
  1.湖南中医药大学，长沙 410208
  2.湖南中医药大学第一附属医院，长沙 410007
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230201
  CLC： R2-0;R22;R285.5;R289;R33
- Published：05 May 2023，
  
  Published Online：05 January 2023，
  
  Received：17 August 2022，
- 稿件说明：
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刘星赐,吴东升,曹晖等.基于LC-MS研究芍药汤对溃疡性结肠炎大鼠粪便代谢产物的影响[J].中国实验方剂学杂志,2023,29(09):61-70.

LIU Xingci,WU Dongsheng,CAO Hui,et al.Effect of Shaoyaotang on Fecal Metabolites in Rats with Ulcerative Colitis Based on Liquid Chromatography-mass Spectrometry[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(09):61-70.
刘星赐,吴东升,曹晖等.基于LC-MS研究芍药汤对溃疡性结肠炎大鼠粪便代谢产物的影响[J].中国实验方剂学杂志,2023,29(09):61-70. DOI： 10.13422/j.cnki.syfjx.20230201.

LIU Xingci,WU Dongsheng,CAO Hui,et al.Effect of Shaoyaotang on Fecal Metabolites in Rats with Ulcerative Colitis Based on Liquid Chromatography-mass Spectrometry[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(09):61-70. DOI： 10.13422/j.cnki.syfjx.20230201.

摘要

目的

基于液相色谱-质谱联用（LC-MS）探究芍药汤对2，4，6-三硝基苯磺酸（TNBS）诱导的溃疡性结肠炎（UC）大鼠的粪便代谢产物的影响。

方法

将SPF级雄性SD大鼠随机分为正常组、模型组、芍药汤组（11.1 g·kg

-1

），除正常组外，采用TNBS诱导UC大鼠模型，除芍药汤组外，各组分别予以生理盐水，均治疗7 d，观察各组大鼠一般情况和疾病活动指数（DAI），采用苏木素-伊红（HE）染色观察结肠组织病理学改变，免疫组化法（IHC）检测结肠组织中白细胞介素-8（IL-8）、白细胞介素-22（IL-22）蛋白表达，采用LC-MS技术检测大鼠粪便样本，并通过主成分分析（PCA）、偏最小二乘法判别分析（PLS-DA）分析数据，对筛人体代谢组数据库（HMDB）数据库及京都基因与基因组百科全书（KEGG）数据库，结合文献筛查，筛选差异代谢物，结合Metabo Analyst 5.0检索，得到通路富集分析。

结果

UC大鼠灌胃芍药汤可使DAI评分降低，与正常组比较，模型组大鼠结肠黏膜结构破坏，黏膜下层炎性细胞浸润，结肠组织IL-8蛋白表达水平显著升高（

0.01），IL-22蛋白表达水平明显升高（

0.05），与模型组比较，芍药汤组大鼠结肠损伤减轻，结肠组织IL-8蛋白表达水平显著降低（

0.01），IL-22蛋白表达水平显著降低（

0.01）。经过筛选处理，与模型组比较，芍药汤组共鉴定出15个差异代谢物，代谢物主要涉及通路包括不饱和脂肪酸的生物合成、亚油酸代谢、萜类骨架生物合成、卟啉和叶绿素代谢、氨基糖和核苷酸糖代谢、嘧啶代谢、类固醇激素生物合成。

结论

芍药汤对UC具有治疗作用，其抗炎作用可能与改善脂质代谢、调节辅因子和维生素代谢、碳水化合物代谢的异常状态有关。

Abstract

Objective

To investigate the effect of Shaoyaotang on fecal metabolites in rats with ulcerative colitis （UC） induced by 2，4，6-trinitrobenzenesulfonic acid （TNBS） based on liquid chromatography-mass spectrometry （LC-MS）.

Method

Male SPF SD rats were randomly divided into normal group， model group and Shaoyaotang group （11.1 g·kg

-1

）. Except for normal group， UC rat model was induced by TNBS， and each group was given normal saline except Shaoyaotang group. All groups were treated for 7 days， and the general condition and disease activity index （DAI） were observed. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes of colon， and the protein expressions of interleukin-8 （IL-8） and interleukin-22 （IL-22） in colon tissue were detected by immunohistochemistry （IHC）. Rat fecal samples were detected by LC-MS， and the data were analyzed by principal component analysis （PCA） and partial least squares discriminant analysis （PLS-DA）. Human Metabolome Database （HMDB） and Kyoto Encyclopedia of Genes and Genomes （KEGG） were searched to screen differential metabolites in combination with literature reference. Then， pathway enrichment analysis was conducted using Metabo Analyst 5.0.

Result

Shaoyaotang （

） decreased the DAI of UC rats. Compared with the normal group， the model group had damaged colonic mucosa structure， submucosal inflammatory cell infiltration， increased protein expressions of IL-8 （

0.01） and IL-22 （

0.05） in colon tissue. Compared with the conditions in the model group， the colonic damage was alleviated in the Shaoyaotang group， and the protein expressions of IL-8 and IL-22 in colon tissue were decreased （

0.01）. After screening， 15 differential metabolites were identified from the Shaoyaotang group， and the involved pathways mainly included biosynthesis of unsaturated fatty acids， linoleic acid metabolism， terpenoid backbone biosynthesis， porphyrin and chlorophyll metabolism， amino sugar and nucleotide sugar metabolism， pyrimidine metabolism and steroid hormone biosynthesis.

Conclusion

Shaoyaotang has a therapeutic effect on UC， and its anti-inflammatory effect may be related to improving lipid metabolism and regulating the metabolism of cofactors and vitamins as well as the abnormal carbohydrate metabolism.

关键词

芍药汤溃疡性结肠炎代谢组学非靶向

Keywords

Shaoyaotangulcerative colitismetabolomicsnon-targeted

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