蜘蛛香提取物治疗腹泻型肠易激综合征模型大鼠粪便UPLC-MS/MS代谢组学研究

Effect of Valeriana jatamansi extract on fecal UPLC-MS/MS metabolomics in rats with diarrheal irritable bowel syndrome

该文研究蜘蛛香提取物对母婴分离联合3种应激所致腹泻型肠易激综合征(IBS-D)模型大鼠的药效作用和干预后粪便中内源性代谢产物的变化,以寻找潜在的生物标志物及相关代谢通路。采用母婴分离联合束缚、冰泳、夹尾3种应激等复制腹泻型IBS动物模型,通过对大鼠的腹部回撤反射压力阈值、粪便含水量、强迫游泳实验不动时长等药效评价指标来考察蜘蛛香提取物各剂量组的治疗作用。在此基础上,收集大鼠粪便,运用UPLC-LTQ-Orbitrap MS平台检测小分子代谢物的代谢轮廓,通过PCA与OPLS-DA等多元统计分析方法寻找差异代谢生物标志物。结果显示,与正常组相比,模型组大鼠腹部回撤反射压力阈值降低、粪便含水量增加、强迫游泳实验的不动时长延长,而经蜘蛛香提取物干预后能使各药效指标显著回调。粪便代谢组学研究结果表明,模型动物经蜘蛛香高剂量组干预后,39个代谢物呈现下调,37个代谢物呈现上调,且均具有显著性差异,进一步分析发现,这些差异代谢物与胆汁酸代谢、不饱和脂肪酸代谢、氨基酸代谢、神经酰胺神经类物质代谢等多种代谢途径相关。该研究证明了蜘蛛香提取物对IBS-D模型大鼠具有明确的药效作用,并从粪便代谢组学的角度探讨了蜘蛛香提取物治疗IBS-D模型大鼠的药效机制。

The purpose of this study was to understand the pharmacodynamic effect of Valeriana jatamansi extract in diarrhea predominant irritable bowel syndrome(IBS-D)rat model induced by maternal separation combined with three kinds of stress,and observe the changes of endogenous metabolites in feces after intervention to find potential biomarkers and related metabolic pathways.The animal model of IBS-D was established by maternal separation combined with restraint,ice swimming and tail clamping.The therapeutic effect of each dose group of V.jatamansi extract was evaluated in terms of abdominal withdrawal reflex pressure threshold,fecal water content and immobility time of forced swimming test.In addition,rat feces were collected for detection of metabolic profiles of small molecular metabolites with UPLC-LTQ-Orbitrap MS platform,so as to find the biomarkers of differential metabolism with multivariate statistical analysis methods such as principal component analysis(PCA)and orthogon partial least squares discrimination analysis(OPLS-DA).The results showed that as compared with the normal group,the threshold of abdominal withdrawal reflex pressure was decreased,the fecal water content was increased,and the immobility time of forced swimming test was prolonged in the model group.The results of fecal metabonomics showed that the levels of 39 metabolites were down-regulated and those of 37 metabolites were up-re-gulated.Further analysis showed that these metabolites were related to bile acid metabolism,unsaturated fatty acid metabolism,amino acid metabolism,ceramide metabolism and other metabolic pathways.This study proved that the extract of V.jatamansi had definite pharmacodynamic effect on IBS-D model rats,and the mechanism was discussed from the perspective of fecal metabonomics.