| The paper took crude angelica sinensis, charred angelica sinensis and alcohol broiled angelica sinensis as study object, and adopted the method of ice water bath injected with epinephrine to establish acute blood stasis rat model. Besides by detecting the indexes of blood rheology and blood coagulation function, the paper evaluated the influence of crude angelica sinensis, charred angelica sinensis and alcohol broiled angelica sinensis on blood stasis rats. The paper, based on Liquid chromatographyflight time-mass spectrometry(LC-Q/TOF-MS) to analyze the metabolomics,the plasma and urine metabolites in each group were detected, the data were analyzed to screen biomarkers with T test of univariate analysis and multivariate analysis of the pattern recognition, applied Met PA database to build differences metabolites related metabolic pathways, Venn diagram analysis was adopted in each group to compare the difference of metabolites, determined the raw angelica, angelica, angelica wine carbon intervention in the metabolism of blood stasis model rats target metabolites, and combined with blood rheology and blood coagulation factor index Pearson correlation analysis, is to explore raw angelica, angelica, angelica wine carbon promoting blood circulation to remove blood stasis and similarities and differences.The main results were as follows:1. Compared with control group, the whole blood viscosity and plasma viscosity of rats in model group increased significantly(P<0.01), and prothrombin time(PT), thrombin time(TT) and part activated clotting time(APTT) live enzymes decreased significantly(P<0.01), FIB increased significantly(P<0.01), 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 alcohol broiled angelica sinensis group and aspirin group decreased significantly(P<0.01), PT, APTT and TT increased significantly(P<0.01), and FIB decreased significantly(P<0.01); the whole blood viscosity and plasma viscosity of rats in rude angelica sinensis and charred angelica sinensis decreased significantly(P<0.05), PT, APTT and TT increased significantly(P<0.05), and FIB decreased significantly(P<0.01). Above results suggesting rude angelica sinensis, charred angelica sinensis and alcohol broiled angelica sinensis could significantly improve the state of blood stasis, blood circulation and removing blood stasis. The curative effect of the alcohol broiled angelica sinensis group was better than that of rude angelica sinensis and charred angelica sinensis.2. Based on Liquid chromatography-flight time-mass spectrometry(LC-Q/TOFMS) to be the metabolomics analysis method,the plasma and urine metabolites in each group were detected, the data were analyzed to screen biomarkers with T test of univariate analysis and multivariate analysis of the pattern recognition. A total of 20 and 79 kind of biomarkers were identified on plasma and urine of blood stasis model group of the rat in positive and negative mode, which is an important evidence to tell whether the blood stasis has occurred.3. Based on Liquid chromatography-flight time-mass spectrometry(LC-Q/TOFMS) to be the metabolomics analysis method,A total of 12 and 13 kind of biomarkers were identified in on plasma and urine of rude angelica sinensis group of the rat, which involve tryptophan metabolism pathways. A total of 19 and 11 kind of biomarkers were identified in on plasma and urine of charred angelica sinensis group of the rat, which involve glycerol phospholipid metabolism and tyrosine metabolism pathways. A total of 18 and 7 kind of biomarkers were identified in on plasma and urine of alcohol broiled angelica sinensis group of the rat, which involve taurine and taurine metabolism, valine, leucine, isoleucine biosynthesis, sphingolipid metabolism, Selenium generation amino acid metabolism, glycolysis and sugar metabolism, and Arginine and proline metabolism pathways. A total of 19 and 28 kind of biomarkers were identified in on plasma and urine of aspirin group of the rat, which involve glutathione metabolism, glycine, serine and threonine metabolism, taurine and hypotaurine metabolism, glycerol phospholipid metabolism, glyceride metabolism, and citric acid cycle pathways.It can be presumed thatrude angelica sinensis,charred angelica sinensis and alcohol broiled angelica sinensis have different treatment effect on blood stasis of rat, which may be related with the amount and type of blood stasis of rats and its intervention with endogenous metabolites as well as metabolic pathways. 4. Venn diagram analysis method was adopted to compare the difference among rude angelica sinensis intervention group,charred angelica sinensis intervention group, alcohol broiled angelica sinensis intervention group, and aspirin intervention group biomarkers with blood stasis of model group rats biomarkers,and screened out target metabolites. The results show that Rude angelica sinensis on blood stasis model of intervention mainly involves the following differences metabolites: four public kinds of biomarkers have Oleic Acid, phosphatidic acid, Cholestane-3,7,12,24,25-pentol, Beta-Guanidinopropionic acid in plasma samples; Seven public kinds of biomarkers have Kinetensin, Metanephrine, Allocystathionine, 5-Hydroxysebacate, L-Kynurenine, Alpha-Linolenoyl ethanolamide, 3-Methylglutarylcarnitine in urine samples.Charred angelica sinensis on blood stasis model of intervention mainly involves the following differences metabolites: eight public kinds of biomarkers have Oleic Acid, Adrenochrome, phosphatidic acid, 5,6-Dihydroxyindole-2-carboxylic acid, Cholestane-3,7,12,24,25-pentol, Lyso PC(P-18:0), Thiethylperazine, Topaquinone in plasma samples; Four public kinds of biomarkers have Kinetensin, Methylglutaric acid,(S)-Methylmalonic acid/Semialdehyde, Alpha-Linolenoyl ethanolamide in urine samples.Alcohol broiled angelica sinensis on blood stasis model of intervention mainly involves the following differences metabolites: six public kinds of biomarkers have Oleic Acid, phosphatidic acid, Cholestane-3,7,12,24,25-pentol, BetaGuanidinopropionic, L-Valine, Guanidine in plasma samples; One public kinds of biomarkers have Alpha-Linolenoyl ethanolamide in urine samples.Aspirin on blood stasis model of intervention mainly involves the following differences metabolites: four public kinds of biomarkers have phosphatidic acid, Hemin, 5,6-Dihydroxyindole-2-carboxylic acid, gangliosides in plasma samples; Eight public kinds of biomarkers have Phenylalanyl-Tyrosine, PC, phosphatidylethanolamine, monoacylglyceride, Allocystathionine, 5-Hydroxysebacate /Adenine, Glutaric acid, Suberylglycine/Isovalerylglutamic acid in urine samples.Venn diagram analysis method was adopted to compare the difference among angelica, angelica charcoal, wine, and metabolites, aspirin group, results of which show that there are differences in the amount and type of target metabolites concerning the birth of angelica sinensis and its processed products intervention blood stasis model rats. |
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