摘要
该研究基于超高效液相色谱-四极杆-飞行时间串联质谱(ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry,UHPLC-Q-TOF-MS)技术研究槐花散对溃疡性结肠炎小鼠的干预及血清代谢物的影响,探究槐花散治疗溃疡性结肠炎的作用机制。采用致炎剂葡聚糖硫酸钠(dextran sulfate sodium salt,DSS)建立溃疡性结肠炎小鼠模型,通过检测疾病活动指数(disease activity index,DAI),结肠表观评价,结肠组织形态和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)等炎症因子的含量,初步评价槐花散对溃疡性结肠炎小鼠的治疗效果。利用UHPLC-Q-TOF-MS技术检测空白组、模型组以及槐花散低、中、高剂量组小鼠血清样本内源性代谢轮廓谱,以主成分分析(principal components analysis,PCA)、偏最小二乘判别分析(partial least squares discrimination analysis,PLS-DA)及正交偏最小二乘判别分析(orthogonal partial least squares discrimination analysis,OPLS-DA)等多元统计分析进行模式识别,以Mass Profiler Professional(MPP)B.14.00软件中fold change≥2且P<0.05为条件筛选潜在生物标志物,并借助MetaboAnalyst 5.0数据库富集代谢通路。结果表明,槐花散可以显著改善溃疡性结肠炎小鼠一般状态,降低DAI评分,改善结肠组织形态,并降低小鼠血清中TNF-α、IL-6和IL-1β的含量。代谢组学结果显示,与槐花散调控相关的潜在生物标志物共38个,主要涉及甘油磷脂代谢,甘氨酸、丝氨酸和苏氨酸代谢,葡萄糖醛酸的相互转化和谷胱甘肽代谢等代谢通路。该研究利用代谢组学分析槐花散治疗溃疡性结肠炎的作用机制,为进一步深入研究奠定基础。
Ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry(UHPLC-Q-TOF-MS)was employed in this study to observe the effect of Huaihua Powder on the serum metabolites of mice with ulcerative colitis and reveal the mechanism of Huaihua Powder in the treatment of ulcerative colitis.The mouse model of ulcerative colitis was established by dextran sodium sulfate salt(DSS).The therapeutic effect of Huaihua Powder on ulcerative colitis was preliminarily evaluated based on the disease activity index(DAI),colon appearance,colon tissue morphology,and the content of inflammatory cytokines such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β).UHPLC-Q-TOF-MS was employed to profile the endogenous metabolites of serum samples in blank control group,model group,and low-,medium-,and high-dose Huaihua Powder groups.Multivariate analyses such as principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA),and orthogonal partial least squares discriminant analysis(OPLS-DA)were performed for pattern recognition.Potential biomarkers were screened by Mass Profiler Professional(MPP)B.14.00 with the thresholds of fold change≥2 and P<0.05.The metabolic pathways were enriched by MetaboAnalyst 5.0.The results showed that Huaihua Powder significantly improved the general state and colon tissue morphology of mice with ulcerative colitis,reduced DAI,and lowered the levels of TNF-α,IL-6,and IL-1βin serum.A total of 38 potential biomarkers were predicted to be related to the regulatory effect of Huaihua Powder,which were mainly involved in glycerophospholipid metabolism,glycine,serine,and threonine metabolism,mutual transformation of glucuronic acid,and glutathione metabolism.This study employed metabolomics to analyze the mechanism of Huaihua Powder in the treatment of ulcerative colitis,laying a foundation for the further research.
作者
韩丽颖
于浩
李天娇
王帅
包永睿
孟宪生
HAN Li-ying;YU Hao;LI Tian-jiao;WANG Shuai;BAO Yong-rui;MENG Xian-sheng(College of Traditional Chinese Medicine,Liaoning University of Traditional Chinese Medicine,Dalian 116600,China;Liaoning Professional Innovation Technology Center for Multidimensional Analysis of Traditional Chinese Medicine,Liaoning Modern Traditional Chinese Medicine Research Engineering Laboratory,Dalian 116600,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2023年第5期1300-1309,共10页
China Journal of Chinese Materia Medica
基金
辽宁省重点研发计划项目(2020JH2/10300088)。