| Blood stasis syndrome(BSS)is a common traditional medical symptom.A variety of diseases are accompanied by the occurrence of BSS,such as unstable angina pectoris,atherosclerosis and myocardial infarction.BSS is characterized by blood flow retardation and blood stagnation,accompanied by hemorheological disorder,vascular endothelial cell damage and inflammation.Furthermore,BSS may result in multiple organ dysfunction in severe state.The treatment of BSS is mainly with Chinese medicine formulations.However,the pharmacodynamic research lacks advanced and scientific theoretical support.Therefore,it is necessary to develop the therapeutic drugs for BSS with clear mechanism of action.As an effective tool for systematic biology research,omics technologies can perform a topdown holistic analysis to complex biological systems,which is consistent with the viewpoint of whole and syndrome in traditional medicine theory.The development of omics technologies has accelerated the diagnosis of complex diseases,realized the transformation of the disease treatment model from one target-one drug to multi-target-multi-component,and promoted the further recognition of Chinese medicine treatment worldwide.Rhodiola wallichiana var.Cholaensis Injection(RWCI)is prepared from the dried roots and stems of RWC in the method of water extraction and alcohol precipitation.There are a variety of pharmacological effects such as activating blood circulation,removing blood stasis and relieving inflammation,which was used to prevent and treat of cardiovascular diseases(CVD)in China.RWCI may play a role in treating CVD by improving BSS,but its mechanism of action needs to be further elucidated.In the present study,the acute blood stasis rat model was established by ice water bath combined with adrenaline.The treatment group was prophylactically given RWCI at different doses.Fecal,plasma,and urine samples were collected for microbiome analysis based on 16 S rRNA high-throughput sequencing and metabolomics analysis based on high-resolution UPLC-Q/TOF-MS techniques.The results showed that there was no significant difference in the diversity and richness of gut microbiota in the control group,model group and treatment group.However,the evenness of gut microbiota was significantly reduced in model group,and it was significantly improved in RWCI treatment group.Compared with the control group,the structure and taxonomic composition of the gut microbiota significantly changed in model group.The relative abundances of Bacteroidetes,Proteobacteria,TM7 and Actinobacteria presented significant difference in model group.However,they were significantly improved in RWCI group.LEfSe analysis and ROC curve verification results showed that RWCI had a significant intervention effect on 3 genera(Bifidobacterium,Enterococcus and Clostridium)and 4 species(Bifidobacterium Pseudolongum,Lactobacillus Helveticus,Lactobacillus Reuteri and Blautia Producta)in the gut microbiota of BSS rats.PICRUSt functional profile prediction showed that the gut microbiota disorder could lead to the changes of the abundance of genes in metabolic pathways and the immune system.Metabolomics studies based on OPLS-DA models identified 14 and 33 differential metabolites in plasma and urine samples,respectively,involving 9 disordered metabolic pathways.Among them,5 metabolic pathways(arachidonic acid metabolism,linoleic acid metabolism,?-linolenic acid metabolism,retinol metabolism and steroid hormone biosynthesis)were significantly related to the occurrence and development of BSS and RWCI intervention.The joint multi-omics analysis was performed based on the microbiome data from 16 S rRNA sequencing and the metabolomics data from UPLC-Q/TOF-MS technology.The results indicated that the changes of the structure and composition of gut microbiota in BSS rats were significantly correlated with arachidonic acid metabolism and steroid hormone biosynthesis pathways.In summary,the structure and composition of the gut microbiota and the metabolic pattern in BSS rats were significantly changed.Moreover,there was a significant correlation between the two variables.RWCI might regulate the metabolism of BSS rats by improving the distribution structure of the gut microbiota. |
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