基于尿液代谢组学研究黄连和胆黄连对热证药效作用机制的差异性

利用超高效液相色谱与LTQ-Orbitrap-MS串联质谱仪的尿液代谢组学方法,研究黄连、胆黄连对热证模型大鼠药效作用机制的差异性,探讨胆黄连炮制的科学性。采用大鼠灌胃附子、干姜、肉桂水煎液15 d并皮下注射干酵母混悬液致热证模型,给药15 d后采集各组大鼠造模后0~6,6~12,12~24 h的尿样;采用主成分分析(PCA)、偏最小二乘-判别分析(PLSDA)法等技术进行数据处理。正常组与模型组在0~6,6~12 h达到分离,12~24 h出现重叠,分离趋势不明显;黄连组和胆黄连组0~6 h大鼠尿样与模型组分离,接近于正常组;黄连组和胆黄连组在0~6,6~12 h大鼠尿样有分离趋势。鉴定30个与热证相关的差异代谢物,结果表明,黄连经猪胆汁炮制后对热证模型大鼠的整体药效作用发生改变,胆黄连解热作用具有多靶点、起效快、作用强度较强的特点,主要通过对胆碱能神经递质、氨基酸代谢、嘌呤代谢的调节发挥解热作用。

Differences in effective mechanisms of Coptidis Rhizoma and bile processed Coptidis Rhizoma on heat syndrome based on urinary metabonomics

A urinary metabonomics method based on ultra-performance liquid chromatography coupled with LTQ-Orbitrap-mass spectrometry (UPLC/LTQ-Orbitrap-MS) was developed to study the difference of action mechanism of Coptidis Rhizoma (CR) and bile processed CR on the heat syndromein rats, and reveal the scientificity of CR processing method. The heat syndrome rat models were established by intragastric administration of water decoction of Aconiti Lateralis Radix Preparata, Zingiberis Rhizoma and Cinnamomi Cortex for 15 days combined with subcutaneous injection of dry yeast suspension. After administration for 15 days, the urine of rats in each group was collected at 0-6 h after modeling, 6-12 h after modeling and 12-24 h after modeling; principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) were used for data treatment. Good separation was observed between the normal groups and model group at 0-6 h and 6-12 h, but overlapped at 12-24 h with no separation trend. Obvious separation was achieved in urine samples between CR group, BCR group and model groups at 0-6 h, close to the normal group. Separation trend occurred between CR group and BCR group at 0-6 h and 6-12 h. Thirty potential biomarkers related with heat syndrome were identified by PLS-DA approach. The results showed that the overall therapeutic effect of CR for heat syndrome had been changed after being processed with pig bile. Bile processed CR has the characteristics of multiple targets, rapid onset and strong effects, mainly playing a role of antipyretic effect through regulating cholinergic neurotransmitter, amino acid metabolism and purine metabolism.