基于UPLC-Q-TOF-MS技术的土家族药三颗针干预溃疡性结肠炎的代谢组学研究

Metabolomics study of Berberidis Radix in intervening ulcerative colitis based on UPLC-Q-TOF-MS

通过代谢组学技术研究土家族药三颗针对葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导的溃疡性结肠炎(ulcerative colitis,UC)小鼠血清、粪便内源性代谢物的影响,探讨三颗针干预UC的代谢途径及可能的作用机制。采用DSS诱导小鼠UC模型。记录小鼠体质量、疾病活动指数(disease activity index,DAI)及结肠长度。采用ELISA法检测小鼠结肠中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素-10(interleukin-10,IL-10)的水平。采用超高效液相色谱-串联三重四极杆飞行时间质谱(UPLC-Q-TOF-MS)技术检测小鼠血清与粪便中内源性代谢物的水平,并结合主成分分析和正交偏最小二乘法判别分析对检测出的差异代谢物进行表征,筛选出潜在的差异代谢物,采用MetaboAnalyst 5.0软件分析可能的代谢通路。结果表明,三颗针可以显著改善UC小鼠的症状,提高组织中抗炎因子IL-10水平;分别在小鼠血清与粪便中鉴定出56、43个差异代谢物,属于脂质、氨基酸和脂肪酸等,经三颗针干预后,发生紊乱的代谢物水平逐渐向正常组靠近。涉及到的代谢通路主要有苯丙氨酸、酪氨酸和色氨酸的生物合成,亚油酸代谢,苯丙氨酸代谢,甘油磷脂代谢等。因此,三颗针可以改善DSS诱导的小鼠UC症状,其机制可能与调节脂质代谢、氨基酸代谢、能量代谢密切相关。

The effect of Tujia medicine Berberidis Radix on endogenous metabolites in the serum and feces of mice with ulcerative colitis(UC)induced by dextran sulfate sodium(DSS)was analyzed by metabolomics technology to explore the metabolic pathway and underlying mechanism of Berberidis Radix in the intervention of UC.The UC model was induced in mice by DSS.Body weight,disease activity index(DAI),and colon length were recorded.The levels of tumor necrosis factor-α(TNF-α)and interleukin-10(IL-10)in colon tissues were determined by ELISA.The levels of endogenous metabolites in the serum and feces were detected by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS).Principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were employed to characterize and screen differential metabolites.The potential metabolic pathways were analyzed by MetaboAnalyst 5.0.The results showed that Berberidis Radix could significantly improve the symptoms of UC mice and increase the level of the anti-inflammatory factor IL-10.A total of 56 and 43 differential metabolites were identified in the serum and feces,respectively,belonging to lipids,amino acids,fatty acids,etc.After the intervention by Berberidis Radix,the metabolic disorder gradually recovered.The involved metabolic pathways included biosynthesis of phenylalanine,tyrosine,and tryptophan,linoleic acid metabolism,phenylalanine metabolism,and glycerophospholipid metabolism.Berberidis Radix can alleviate the symptoms of mice with DSS-induced UC,and the mechanism may be closely related to the regulation of lipid metabolism,amino acid metabolism,and energy metabolism.