基于UPLC-Q-TOF-MS技术白术多糖干预溃疡性结肠炎的代谢组学研究

Metabolomic study of Atractylodes macrocephala polysaccharide intervention in ulcerative colitis based on UPLC-Q-TOF-MS technology

目的研究白术Atractylodes macrocephala多糖对溃疡性结肠炎(ulcerative colitis,UC)小鼠的影响,探讨白术多糖干预UC的代谢途径及可能的作用机制。方法采用葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导小鼠UC模型,给予白术多糖或美沙拉嗪干预后,测定小鼠体质量、疾病活动指数(disease activity index,DAI)评分及结肠长度;采用ELISA检测小鼠血清中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)水平及超氧化物歧化酶(superoxide dismutase,SOD)、髓过氧化物酶(myeloperoxidase,MOP)活性;采用苏木素-伊红(HE)染色观察结肠组织病理变化;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)检测小鼠血清内源性代谢物水平,并结合主成分分析和正交偏最小二乘法判别分析对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网址分析可能的代谢通路。结果白术多糖显著改善UC小鼠的症状和结肠组织病理损伤,提高血清中IL-10水平和SOD活性(P<0.05、0.01),降低TNF-α水平和MPO活性(P<0.05、0.01),并通过α-亚麻酸代谢通路、类固醇激素生物合成通路、初级胆汁酸生物合成通路、甘油磷脂代谢、泛酸和辅酶A生物合成通路、不饱和脂肪酸生物合成通路等代谢途径回调UC小鼠的异常代谢产物。结论白术多糖具有防治UC的作用,其机制可能与平衡促炎因子与抗炎因子、增强抗氧化和调节脂质代谢等通路相关。

Objective To study the effects of Atractylodes macrocephala polysaccharide(AMP)on ulcerative colitis(UC)mice,and investigate the metabolic pathways and possible mechanisms of AMP intervention in UC.Methods A mice UC model was induced with dextran sulfate sodium(DSS),after the intervention of AMP or mesalazine,body weight,disease activity index(DAI)score and colon length were recorded;Levels of tumor necrosis factor-α(TNF-α),interleukin-10(IL-10)and activities of superoxide dismutase(SOD),myeloperoxidase(MOP)in serum of mice were measured by ELISA;HE staining was used to observe the pathological changes of colon tissue;The levels of endogenous metabolites in serum of mice were detected by ultra performance liquid chromatography-tandem triple quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS),the differential metabolites were characterized by principal component analysis and orthogonal partial least squares-discriminant analysis,and the potential differential metabolites were screened out.The website of MetaboAnalyst 5.0 was used to analyze the possible metabolic pathways.Results AMP significantly improved the symptoms and pathological damage of colon tissue in UC mice,increased IL-10 level and SOD activity in serum(P<0.05,0.01),decreased TNF-α level and MPO activity(P<0.05,0.01),modulated the abnormal metabolites in UC mice byα-linolenic acid,steroid hormone biosynthesis,primary bile acid biosynthesis,glycerophospholipid metabolism,pantothenic acid and coenzyme A biosynthesis and unsaturated fatty acid biosynthesis metabolic pathways.Conclusion AMP has a preventive and curative effect on UC,and its mechanism may be related to balancing pro-inflammatory factors and anti-inflammatory factors,enhancing antioxidation and regulating lipid metabolism.