| Notoginseng Radix et Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma is a classic clinical herbal pair for the treatment of blood stasis syndrome and the main component of various herbal compound preparations for promoting blood circulation and removing blood stasis on the market.Currently,the research on this herbal pair mainly focuses on the changes of chemical components before and after compatibility,the content changes of blood-activating components in different proportions,the mechanism of anti-thrombosis and inhibition of platelet aggregation.And the establishment method of blood stasis model is mainly based on ’drug-environmental stimulation’.The research on blood stasis caused by external force mainly focuses on simple pharmacodynamics and modeling methods.So,this study reolicated the model of traumatic blood stasis and compared the improvement effects of this herbal pair(3:1,1:1,1:3)on hemorheology,swelling of injured limbs and muscle damage on rats with traumatic blood stasis.The plasma and urine metabolomics study,identification of chemical components and metabolites in vivo and network pharmacology study of the herbal pair promoting blood circulation and removing blood stasis were carried out based on UPLC-Q-TOF-MS/MS and bioinformatics database.The promoting blood circulation and removing blood stasis mechanism of this herbal drug was explored from the perspective of metabolism,pharmacodynamic material basis and network pharmacology.The results of the study are as follows:(1)After reproduction of model of traumatic blood stasis,the positive drug,Notoginseng Radix et Rhizoma,Salviae Miltiorrhizae Radix et Rhizoma and the Notoginseng Radix et Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma(3:1,1:1,1:3)were selected to study the effect of promoting blood circulation and removing blood stasis.The results showed that they can all reduce the hemorheological indexes and the difference between daily and initial leg diameter of the injured limb were significantly and improve muscle tissue injuries,comparing to the model group.After recovery,the herbal pair 3:1 has the best therapeutic effect.(2)In the metabolomics research,the UPLC-Q-TOF-MS/MS technology was used to screen and analyze the differential metabolites and metabolic pathways in plasma and urine of rats with traumatic blood stasis and compare the callback and intervention effect of the positive drug,Notoginseng Radix et Rhizoma,Salviae Miltiorrhizae Radix et Rhizoma,the 3:1,1:1,1:3 herbal pair on differential metabolites and metabolic pathways.The results showed that 22 plasma differential metabolites were identified in plasma,leading to the disorder of linoleic acid metabolism,alpha-linolenic acid metabolism,arachidonic acid metabolism,steroid hormone biosynthesis and alanine,aspartate and glutamate metabolism.And 12 urine differential metabolites were identified,leading to the disorder of phenylalanine metabolism,steroid hormone biosynthesis,pentose and glucuronic acid conversion and lysine degradation.After administering positive drug,Notoginseng Radix et Rhizoma,Salviae Miltiorrhizae Radix et Rhizoma and the 3:1,1:1,1:3 herbal pair,there were 14,11,11,17,15 and 13 plasma differential metabolites were called back,respectively regulating 4,3,4,5,4 and 4 plasma-related metabolic pathways.And there are 9,6,6,9,8 and 8 urine differential metabolites were called back,respectively regulating 4,2,2,4,3 and 2 urine-related metabolic pathways.The herbal pair 3:1 recalled the most differential metabolites in plasma and urine and regulated the most related metabolic pathways.(3)In identification of chemical constituents and metabolites of herbal pair(3:1),22 prototype components and 13 metabolic components were identified in plasma.And 13 prototype components and 9 metabolic components were identified in urine.The saponins of Notoginseng Radix et Rhizoma,the water-soluble phenolic acids and the fat-soluble tanshinones of Salviae Miltiorrhizae Radix et Rhizoma can be detected in the form of prototypes and metabolites in the plasma and urine of rats with traumatic blood stasis.(4)In the network pharmacology research,the PubChem,SwissTargetPrediction,GeneCards,OMIM and DrugBank databases are used to predict and screen the targets of differential metabolites,chemical components in vivo,metabolites and disease,and take targets intersections.Subsequently,protein-protein interaction network were constructed through Cytoscape software.Meanwhile,KEGG and GO enrichment analysis of intersection targets was performed with Metascape database.As a result,a total of 114 intersection targets were obtained,173 pathways were enriched in KEGG,and 1569 entries were enriched in GO,including 1359 biological process entries,76 cellular composition entries and 134 molecular function entries.KEGG and GO enrichment mainly involve arachidonic acid metabolism,complement and coagulation cascade,response to injury and response to inflammatory.To sum up,according to improvement of hemorheology,swelling of injured limbs,muscle damage and the results of plasma and urine metabolomics,it was found that the 3:1 herbal pair had the best effect of promoting blood circulation and removing blood stasis.Based on identification results differential metabolites,chemical components,metabolites in vivo and results of network pharmacology research,the ’componenttarget-pathway’ network of this herbal pair promoting blood circulation and removing blood stasis was revealed initially.The herbal pair of Notoginseng Radix et RhizomaSalviae Miltiorrhizae Radix et Rhizoma can intervenes in inflammation-related pathways such as arachidonic acid metabolism,linoleic acid metabolism and αlinolenic acid metabolism by adjusting the content of differential metabolites about traumatic blood stasis and regulating blood stasis-related targets,so as to play the role of promoting blood circulation and removing blood stasis.It provides a theoretical reference for the clinical application and further development of this herbal drug. |
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