Study On The Bioactive Substances Of Jun-Shi Couple Herbs In Salvia Miltiorrhiza-Dalbergia Odorifera

Investigation on the compatibile regulation is a hot and difficult topic in the research area of traditional Chinese medicine. Couple Herb is turn between single herb and complex prescription. Jun-Shi couple herb is the special compatibility consists of Jun herb and Shi herb in a complex prescription. It not only displays the 'Jun, Chen, Zuo, Shi' compatibility of a complex prescription and 'Qi Qing' synergism, but also has the properties of simple composition and easy to research process. Thus, the investigation on synergitic action of Jun-Shi couple herb could elucidate the compatible mechanism of complex prescription. Meanwhile, it also plays an important role in enriching and developing the compatible theory of complex prescription and development of innovative medicine. In this thesis, Salvia milltiorrhiza-Dalbergir odorifera (SM-DO) was selected as the research target, and Danshensu (DSS), protocatechuic acid (PA), protocatechuic aldehyde (PAH) was used as markers to study the effects of DO on the Pharmacokinetics and bioactive substances in vivo in SM. The main content of this thesis was showed as follows:1. A new high performance liquid chromatography (HPLC) method for the analysis of DSS, PA and PAH of SM in plasma is established. The results show that the detection limits of the above substances are 0.031, 0.008, 0.004μg/mL. The linear range of DSS, PA and PAH are 0.031-4.0μg/mL, 0.016-2.0μg/mL, 0.008-1.0μg/mL, respectively. The extracting recovery of low, medium and high concentration are among the range of 88.62-95.80%, and recovery of the proposed method are 86.26-98.40%. The relative standard deviation (RSD) of inter- and intro-day precision and stability are in accord with the requirements of biological sample analysis of Food and Drug Administration. All the above results demonstrated that the proposed method could be used in pharmacokinetic research of DSS, PA and PAH.2. The pharmacokinetic process of DSS and PA standards are investigated by the above method. Based on this, the pharmacokinetic differences of DSS and PA are analyzed by the proposed method after intragastric administration of DSS, PA, DSS-DO, PA-DO, SM and SM-DO. The results show that the pharmacokinetic process of DSS and PA are confirmed to a two compartment open model. In addition, there are substances in SM that could accelerated the absorption rate, increase the bioavailabilty of DSS. Shi herb (DO) could decelerate the aborption rate, increase the amount in blood, increase the bioavailability, decelerate excretion and prolong the absorption time of DSS and PA standard and DSS and PA in Jun herb (SM). These results have verified the action of enhancing therapeutic effect and guidance of DO, which provided the evidence to elucidate the synergitic action in vivo of SM-DO.3. The obtained chromatographic data are calculated by principal components analysis and Granger causality analysis, the results show that NM19, ES21, ES23, ES32 and ES34 are the key substance led to the obvious change of chromatograms. Based on this, the retention characteristic of NM19, ES23 and ES32 onβ₂-AR biochromatographic coulumn are investigated, and the results demonstrate that those three substance has specific interaction withβ₂-AR. Moreover, high performance liquid chromatography- mass spectrometry are used to identify NM19, ES23 and ES32, the results show that the molecular weight of ES23 and ES32 are 204.2 and 179.0, respectively. Thus, ES23 and ES32 are identifed as L-tryptophan and hippuric acid.