基于非靶代谢组学研究痰热腑实证型出血性中风后发生二次出血的生物标志物  

Study on the Biomarkers for Secondary Hemorrhage after Hemorrhagic Stroke of Phlegm-Heat and Bowel Excess Pattern Based on Non-Targeted Metabolomics

作　　者：邓娟 郭凯欣 柯雪红[2] 陈为[2] DENG Juan;GUO Kaixin;KE Xuehong;CHEN Wei(Guangzhou University of Chinese Medicine,Guangzhou 510405,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]广州中医药大学,广东广州510405 [2]广州中医药大学第一附属医院,广东广州510405

出　　处：《中医药学报》2025年第4期62-70,共9页Acta Chinese Medicine and Pharmacology

基　　金：国家自然科学基金项目(82074099)。

摘　　要：目的:通过非靶代谢组学研究痰热腑实证型出血性中风后发生二次出血的生物标志物,并筛选出具有预测二次出血价值的生物标志物,为今后临床预测疾病发生提供新的参考指标。方法:收集广州中医药大学第一附属医院神经外科临床西医诊断为高血压脑出血,中医诊断为痰热腑实证型出血性中风患者共60例,其中仅发生一次出血、发生二次出血患者各30例;同时招募30例健康志愿者作为空白对照。应用QExactive台式四极杆-轨道阱高分辨质谱仪(UHPLC-Q-Orbitrap HRMS)技术对一次脑出血(NCX)、二次出血(NCX+)及健康组(CON)之间血浆差异代谢物进行鉴定。采用主成分分析(PCA)、正交偏最小二乘法判别分析(OPLS-DA)、差异物的热图制作、ROC曲线绘制,对代谢谱数据进行多维统计分析,筛选出具有预测二次出血潜力的生物标志物。结果:健康组/一次出血组差异代谢物9种,健康组/二次出血组差异代谢物共22种,一次出血/二次出血组差异代谢物共11种。DL-甲羟戊酸、2-叔丁基-4,6-二硝基苯酚、4-庚基苯酚、8-甲基-2(1H)-喹啉酮、5-羟基吲哚-3-乙酸、L-犬尿氨酸、苏氨酸ROC曲线下面积分别为0.95、0.97、0.98、0.96、0.97、0.97、0.85。色氨酸代谢通路P值为0.002,Impact值为0.12;缬氨酸、亮氨酸和异亮氨酸降解通路P值为0.07,Impact值为0.03。结论:DL-甲羟戊酸、2-叔丁基-4,6-二硝基苯酚、4-庚基苯酚、8-甲基-2(1H)-喹啉酮、5-羟基吲哚-3-乙酸、L-犬尿氨酸、苏氨酸7种差异代谢物均具有预测痰热腑实证型出血性中风后发生二次出血的潜力。色氨酸代谢为显著差异代谢通路。Objective:To explore the biomarkers for secondary hemorrhage after hemorrhagic stroke of phlegm-heat and bowel excess pattern using non-targeted metabolomics and identify biomarkers with predictive value for secondary hemorrhage,providing new reference indicators for clinical prediction of disease occurrence.Methods:Sixty patients diagnosed with hypertensive cerebral hemorrhage by Western medicine and phlegm-heat and bowel excess pattern by Chinese medicine were selected from the Department of Neurosurgery,The First Affiliated Hospital of Guangzhou University of Chinese Medicine from June 2021 to April 2023.They were divided into two groups:those who had a single hemorrhage(30 patients)and those who experienced secondary hemorrhage(30 patients).Additionally,thirty healthy individuals were recruited as controls.The UHPLC-Q-Orbitrap HRMS technique was used to identify differential metabolites in plasma between one-time hemorrhage(NCX),secondary hemorrhage(NCX+),and healthy groups(CON).Multidimensional statistical analyses,such as Principal Component Analysis(PCA),Orthogonal Partial Least Squares Discriminant Analysis(OPLS-DA),heatmap of differential metabolites,and ROC curve plotting,were employed to select potential biomarkers for predicting secondary hemorrhage.Results:There were 9 differential metabolites between the healthy group and the one-time hemorrhage group,22 between the healthy group and the secondary hemorrhage group,and 11 between the one-time and secondary hemorrhage groups.Biomarkers such as DL-methylmalonic acid,2-tert-butyl-4,6-dinitrophenol,4-heptylphenol,8-methyl-2(1H)-quinolinone,5-hydroxyindole-3-acetic acid,L-canavanine,and threonine showed area under the curve(AUC)values of 0.95,0.97,0.98,0.96,0.97,0.97,and 0.85,respectively.The tryptophan metabolism pathway had a significant difference with a p-value of 0.002 and an Impact value of 0.12;valine,leucine,and isoleucine degradation pathways showed a p-value of 0.07 and an Impact value of 0.03.Conclusion:Seven differential metabolites,DL-methylm

关 键 词：出血性中风 痰热腑实 二次脑出血 代谢组学 

分 类 号：R284.1[医药卫生—中药学]