“冬病夏治”方药穴位贴敷治疗哮喘豚鼠的尿液代谢组学研究

Metabonomic Analysis of Urine from Bronchial Asthma in Cavia Cobaya Treated by Acupoint External Application with ‘Treating Winter Disease in Summer’Prescription

目的:采用代谢组学方法,研究"冬病夏治"方药穴位贴敷法治疗哮喘豚鼠的机制。方法:通过超高效液相色谱结合高分辨率质谱联用(UPLC-Q-TOF-MS)技术分析空白组、卵蛋白致敏的豚鼠哮喘模型组及中药穴位贴敷组豚鼠尿液代谢物图谱.使用SIMCA-P+软件正交偏最小二乘判别分析法(PLS-DA)对采集的数据进行处理。结果:哮喘模型组和空白组、穴位给药组与哮喘模型组分别呈现聚类分布,2组之间未见任何交叉与重叠,得到了明显区分。初步确认氮4-乙酰基胞苷(N4-acetylcytidine,ac4c)、6-甲基腺苷(6-methyladinosinek,m6a)以及未确定化合物(荷质比为365.14)为哮喘发作潜在的生物标志物。甲基戊二酸、1-甲基次黄苷为冬病夏治方治疗哮喘潜在的生物标志物。结论:本研究建立了基于豚鼠尿液的UPLC-Q-TOF-MS代谢组学研究方法,初步阐释了"冬病夏治"方药治疗哮喘的代谢组学机制。研究结果对用现代生物科学技术解释传统中药制剂理论有一定的启发作用。

Objective： To study the mechanism of acupoint external application with ＂treating winter disease in summer＂prescription on bronchial asthma in cavia cobaya.Method： The metabolite profiles of the urine samples of the blank control group,the asthma-model group and the acupoint administration group were detected by ultraperformance liquid chromatography coupled with Q-TOF mass spectrometry（UPLC-Q-TOF-MS）.The data was analyzed by partial least square discriminatory analysis（PLS-DA） in SIMCA-P ＋.Result： The result showed that metabolite profile of the urine sample was separated obviously and had no any overlapping between the asthma-model group and the blank control group,the acupoint administration group and the asthma-model group.N4-acetylcytidine,6-methyladinosinek and an undetermined compound were confirmed initially as the marker of asthma,while methylglutarate and methyl inosine,the markers of the ＇treating winter disease in summer＇prescription.Conclusion： The topic establishes a method of metabonomic analysis for urine from cavia cobaya using UPLC-Q-TOF-MS.The study shows the mechanism of metabolism for the therapy of asthma with＇treating winter disease in summe＇prescription.The results will be of great helpful to explain the theory of Chinese drugs pharmaceutics using biological science of modern time.