Study On Substantial Basis Of Effects Of Jun-Shi Couple Herb Of Salvia Miltiorrhiza-Dalbergia Odorifera

Fufang, as the main pattern of prescribed Chinese medicines, reflects the key point of Chinese medical philosophy that determines the treatment based on differentiation of symptoms and signs. Study on substantial basis of effects of Fufang is the hot and difficult topic in the field of traditional Chinese medicine. Couple Herb which is the connection between Fufang and single herb is simple and easy to study. Jun-Shi Couple Herb is a special kind of Couple herb which is formed by Jun herb and Shi herb in a Fufang, and reflects the synergistic interaction of Fufang. Metabolomics is a newly emerging subject whose aim is to quantitatively describe the dynamic change of metabolites in biosystem. Its core idea of research coincides with that of traditional Chinese medicine perfectly, so metabolomics has gained unprecedented development in the field of traditional Chinese medicine in recent years.In this thesis, Salvia milltiorrhiza-Dalbergir odorifera (SM-DO) was selected as the research target, and danshensu, protacatechuic acid, protocatechuic aldehyde were selected as research index. With metabolomics as the main medium, the synergistic effect of DO on the pharmacokinetics and substantial basis in SM in SD rats was investigated. The main contents and results of this thesis were as follows:1. A new high performance liquid chromatography (HPLC) method for analysis of DSS, PA and PAH simultaneous in rat plasma was established. The results showed that the detective limits of these three substances were 0.25,0.2 and 0.125μg/mL, and the linear ranges of these three substances were 250～2.5,200～2.0,125～1.25μg/mL respectively. The extracting recovery of low, medium and high concentration were 91.17～97.02%, while method recovery were 81.39～97.66%. The relative standard deviation (RSD) of inter-and intro-day precision and stability were to accord with the requirments of biological samples analysis. And the constituents of SM and endogenous substances were separated completely. The results above indicated that this method was suitable for the investigation of DSS, PA, and PAH in rat plasma, and provided a sensitive analytic method for the study of substantial basis of effects of Jun-Shi Couple Herb of SM-DO.2. Influence of Shi Herb DO on SM's pharmacokinetic process in nomal rat was investigated by the above method. Principal components analysis (PCA) and SPSS correlation analysis were used to handle the data. The results were as follows:①DO could accelerate the absorption rate of DSS, while slow down the rate of clearance of DSS and PA. So DO could prolong the residence time of DSS and PA in the living organism and enhance their therapeutic effect, which provided evidence to elucidate the synergitic effect in vivo of SM-DO.②PAH of SM could transform to PA in vivo by specific physiological route, then part of PA produced two metabolites which named NM28 and NM29.③Results of PCA and SPSS correlation analysis showed that NM28, NM29 and tryptophane were the main reasons resulting in the significant differences between nomal single group and nomal combined group. In another word, these substances may related to the substantial basis of effects of Jun-Shi Couple Herb of SM-DO.3. The pharmacokinetic process of DSS and PA in qi-stagnancy and blood stasis rats was investigated after orally adminitrated SM and SM combined with DO. Principal components analysis (PCA) and SPSS correlation analysis were used to handle the data. The results were as follows:①Compared with model single group, DO could accelerate the absorption rate of DSS and PA significantly, while slow down their rate of clearance. At the same time, the concentration of DSS and PA decreased in plasma.②In model combined group, two new metabolites, NM30 and NM38, were observed besides NM28 and NM29.③Results of PCA and SPSS correlation analysis showed that NM28, NM29, NM38 and tryptophane were the main reasons resulting in the significant differences between model single group and model combined group. In another word, these substances may related to the substantial basis of effects of Jun-Shi Couple Herb of SM-DO.4. The solution of DSS, PA and PAH standards were orally adminitrated to SD rats and plasma was collected in laying time points. Results indicated that PAH of SM transformed to PA, then part of PA transformed to NM28 and NM29 quickly. High performance liquid chromatography-mass spectrometry were used to identify metobolites, the results showed that the molecular weight of NM28 and NM29 are 387.4 and 431.3 respectively.