| Taohongsiwutang is a classical prescription on promoting blood circulation. The use of Taohongsiwutang is more and more widely because of its function to invigorate blood circulation. In this paper, with Taohongsiwutang to be the research object and liquid chromatography-flight time-mass spectrometry (LC-Q/TOF-MS) to be the analysis method, the intervene mechanism of Taohongsiwutang on blood stasis model was studied and the metabolic network of Taohongsiwutang on blood stasis rats was established with network analysis. These researches lay the foundation of interpreting the blood activating and stasis removing effect of Taohongsiwutang, and provided new thoughts and methods for studying the effect of traditional Chinese medicine comprehensively. The main results were as follows:1. T he acute bl ood stasis rat model was establ ished by combi nati on use of ice water bath and injected with epinephrine. By detecting the indexes of blood rheology and blood coagulation function, the influence of Taohongsiwutang on blood stasis rats was evaluated. Compared with control group, the whole blood viscosity and plasma viscosity of rats in model group increased significantly-1 P<0.05-1, indicating the blood stasis rat model could be established with this method. Compared with model group, the whole blood viscosity and plasma viscosity of rats in Taohongsiwutang group and aspirin group decreased significantly-1 P<0.05-1, suggesting Taohongsiwutang and aspirin could significantly inproved the state of blood stasis. Compared with normal group, prothrombin time (PT), thrombin time (TT) and part activated clotting time (APTT) live enzymes were significantly reduced-1 P<0.01-1. Compared with model group, PT↑↑TT↑↑APTT and fibrinogen(FIB) contentin Taohongsiwutang group had the trend to close to normal group, suggesting Taohongsiwutang could improve the status of acute blood stasis when the body was in hypercoagulable state.2. Based on LC-Q/TOF-MS metabolomics, the urine metabolites in all groups were detected, and the data were analyzed with principal component analysis(PC A) and partial least squares discriminant analysis (PLS-DA) method. The results showed that the normal group and model group could be completely separated, and Taohongsiwutang group and aspirin group showed a trend to near normal group. According to VIP>1.0 and P<0.05, a total of 53 different metabolites were identified. Then, the identified 53 metabolites were screened with Cytoscape and 5 biomarkers were obtained. These 5 biomarkers were mainly related to cysteine and methionine metabolism, phenylalanine metabolism and tyrosine metabolism. With the intervene of Taohongsiwutang and aspirin, the levels of these biomarkers showed a trend to back to normal group, and the disordered metabolism pathways were improved significantly.3. Based on LC- Q/TOF-MS metabolomics, the plasma metabolites in all groups were detected, and the data were analyzed with principal component analysis(PC A) and partial least squares discriminant analysis (PLS-DA) method. The results showed that the normal group and model group could be completely separated, and Taohongsiwutang group and aspirin group showed a trend to near normal group. According to VIP>1.0 and P<0.05, a total of 56 different metabolites were identified and 7 biomarkers were obtained with the same method. These 7 biomarkers were mainly related to arachidonic acid metabolism, citric acid cycle and glutamate and glutamine metabolism. With the intervene of Taohongsiwutang and aspirin, the levels of these metabolites showed a trend to return to normal group, and the disordered metabolism pathways improved significantly.To sum up, Taohongsiwutang mainly invigorate the urine by cysteine and methionine metabolism, phenylalanine metabolism and tyrosine metabolism, and the plasma by arachidonic acid metabolism, citric acid cycle and glutamate and glutamine metabolism. In this paper, the metabolomics was used to investigate the internal mechanism of Taohongsiwutang of promoting blood circulation and removing blood stasis, which provided a scientific method for mechanism research, exploitation and appl i cati on of Taohongsi wutang. |
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