Exploring the mechanism of Danggui-Guizhi intervention on cold coagulation blood stasis syndrome based on dynamic urine metabolomics

Objective: To explore the dynamic regulatory mechanism of Danggui-Guizhi(DG) intervention on cold coagulation blood stasis syndrome(CCBSS) by employing 1H NMR dynamic urine metabolomics technology. Methods: Fifteen female SD rats were randomly divided into normal group, model group and DG 3∶1 group.Weight changes, hemorheology, and fibrinogen(FIB) indicators were used to evaluate the success of the model construction. Meanwhile, urine samples were collected from each group of rats at different time points(day 0,day 5, day 10, and day 14) for 1H NMR metabolomics analysis, so as to elucidate the pathogenesis of CCBSS and the dynamic regulatory mechanism of DG intervention on CCBSS at metabolic level. Results: Compared with the model group, the rats in the DG 3∶1 group exhibited a deceleration in weight loss, and FIB indicators underwent a call-back(all P>0.05) after the DG intervention. Besides, there was a significant call-back in the hemorheological indicators(P<0.01). The dynamic urine metabolomics analysis revealed that there was a significant distinction in the metabolic profile between the model group and the normal group on days 5, 10, and 14,and the metabolic profile of the DG 3∶1 group was gradually separated from the model group and then approached the normal group as time progressed. Based on multivariate statistical analysis, a total of 18 biomarkers associated with CCBSS were identified, which involved metabolic pathways including taurine and hypotaurine metabolism. The DG 3∶1 group could call back the disorder of these 9 biomarkers(lactate, pyridoxine, succinate,taurine, creatine phosphate, hippurate, 4-hydroxyphenylacetate, 2-oxoglutarate, citrate)(all P<0.05), which mainly interfered with the disorders of taurine and hypotaurine metabolism, as well as TCA cycle and other pathways. Conclusion: The occurrence and development of CCBSS is a dynamic and slow process, and DG 3∶1can improve this syndrome by calling back various metabolites at different time points.