血竭活血化瘀作用的GC-MS代谢组学初步研究

目的：利用代谢组学技术，从内源代谢产物的角度探讨传统药材—血竭的活血化瘀作用。方法：将大鼠分组，分别为正常对照组、血瘀证模型组（血瘀证模型大鼠）和血竭给药组（血瘀证大鼠灌胃给予血竭），采集血浆并采用GC-MS 方法检测各组大鼠血浆中的内源代谢产物。通过主成分分析和偏最小二乘法等数据分析方法寻找血瘀证大鼠的血浆的差异代谢物，分析大鼠给药后血浆内源代谢产物的变化趋势。结果：建立了基于GC-MS 的大鼠血浆内源性代谢产物分析方法，并进行了方法学的确证；采用该方法在大鼠血浆中分析得到约200个数据点，其中约有30个数据点在与NIST 库的匹配中得到了初步的鉴定。通过多元数据分析发现了若干血瘀证大鼠差异代谢物，包括乳酸、D-3-羟基丁酸、缬氨酸、异亮氨酸等，这些差异性代谢物与血瘀证相关的代谢通路有密切的联系；血瘀证大鼠经过血竭给药之后，其血浆中的差异代谢物的浓度发生明显的变化，即各差异代谢物指标与血瘀证模型组相比都有一定的恢复作用，代谢组学研究结果与血瘀证大鼠血液流变学实验结果相吻合。结论：建立了血瘀证大鼠的血浆内源代谢产物的研究方法，并将该方法应用于血竭活血化瘀作用的研究中，从代谢组学的角度证实了血竭对血瘀证的治疗作用。

Objective: To discuss the effect of promoting blood circulation and removing blood stasis of Daemonorops draco Bl., which is a traditional Chinese medicine, in the perspective of metabolism.Methods: Endogenous metabolites of the control group, blood stasis model group and dragon's blood.administration group were determined by using GC-MS analysis and compared.Statistical analyses, such as principal component analysis (PCA) and partial least squares method (PLS), were used for discovering rat plasma biomarkers, and the changing tendencies of plasma endogenous metabolites were analyzed after the dragon's blood.administration.Results: An analytical method of rat plasma endogenous metabolites was developed based on GC-MS.Method validation was also carried out.Approximate 200 data points were obtained by the developed method, among which, approximate 30 data points can be preliminarily identified by matching the NIST database.Several potential rats biomarkers of blood stasis syndrome were discovered, including lactic acid, D-3-hydroxy butyric acid, valine, isoleucine and so on, which are closely related to the metabolic pathways including energy metabolism, oxidative stress and γ-glutamyl cycle.The changing tendencies of these putative biomarkers were quantified after the dragon's blood. administration, and results showed that the indexes had a certain role in the recovery compared to those in the model group, and the results were consistent with blood rheology study.Conclusion: The investigation method of rat plasma endogenous metabolites was developed.The method was applied to study the effect of promoting blood circulation and removing blood stasis of dragon's blood.Therapeutic effect of dragon's blood.on blood stasis syndrome was proved from the perspective of metabonomics.