Study On The Spectrum - Effect Relationship And Interaction

Corydalis yanhusuo W.T. Wang, commonly known as yuan hu, is one of the famous "Zhejiang Bawei", beginning as set out in "bencao shiyi". In Chinese pharmacopoeia, source of quality goods is dry tubers of poppy (Papaveraeeae) purple pansy genera (Corydalis) yanhusuo plants(Corydalis yanhusuo w. t. Wang).It is the traditional Chinese medicine for a line spirit blood circulation and relieving pain.Its application has a long history. It has a significant role in many aspects for the central nervous system and cardiovascular system.So far, study on yanhusuo is confined to the tetrahydropalmatine or its derivatives of tertiaryamine alkaloids, while the study of quaternary ammonium alkali ingredients is less reported, nor can the correlation of different kinds of alkaloids with their efficacy and interaction research be found. Therefore, this paper studies focusing on fingerprint-efficacy study of quaternary alkaloids and interaction between different alkaloids.In this dissertation, by the application of high performance liquid chromatography (HPLC) technique,40different species and habitat of yanhusuo medicinal materials were analyzed, and a fingerprint analysis method was established.Firstly, the national pharmacopoeia similarity analysis software was applied to HPLC similarity analysis, and ten common peak was found preliminary. Separation, preparation, recognition and identification of the common peak was carried out through the guidance of the fingerprint and chemical separation methods (such as macroporous resin, silica gel, MCI).By UPLC analysis method, this dissertation optimized the conditions of liquid phase analysis, established UPLC fingerprint of different rhizoma Corydalis alkaloid, and carried on the methodology. As a result, the fingerprint of37different species and habitat of yanhusuo were compared, their fingerprint common model was set up, and peak area of37samples of ten common peak were analyzed quantitatively. According to the peak area, the matlab software was used for similarity analysis and cluster analysis. The results showed that quality difference of different species and habitat of yanhusuo was very large, with the sample purchased from Beijing tongrentang (producing area of Zhejiang) as reference. We found that Hunan vinegar rhizoma Corydalis with producing area of Zhejiang, Sichuan, was close and their similarity degree was higher, so they could get together for a class.Shanxi and Shandong raw product had the furthest distance from the reference.UPLC-Q-TOF was applicatied to online quickly identify the main chemical composition of tertiary and quaternary alkaloids. In this dissertation, through the method of cell survival rate determined by MTT,37kinds of different species and habitat of quaternary alkaloids from Corydalis yanhusuo W.T. Wang were compared on H9C2cardiac muscle cell hypoxia/reoxygenation injury protection. Each sample was choosed for three different concentrations to receive the results of pharmacodynamics.This paper applicated methods of multiple regression analysis, grey relational analysis and correlation analysis to study fingerprint-efficacy which combined chemical fingerprint information with pharmacodynamic results of different regions of37kinds of rhizoma Corydalis quaternary ammonium alkali. Results of these three methods were consistent in general, supported by each other. The results showed that dehydrocorybulbine violet peak4with the largest contribution to the efficacy of activity, was the important one in Corydalis quaternary ammonium alkali active substances. Peak3(13-Methyldehydrocorydalmine), peakl (columbamine), peak8(dehydrocorydaline) and peak6(palmatine) were also positive on pharmacodynamic activity. These six peaks with positive contribution to the activity of efficacy together constituted the rhizoma Corydalis quaternary ammonium alkali medicinal material group against myocardial ischemia. Peak5(13-Methylpalmatrubine) and peak7(berberine) were very low or negative for efficacy. This study "fingerprint-efficacy relationship"for quality control model of traditional Chinese medicine provided theoretical basis and practical exploration.By established pharmacodynamic evaluation method, the EC50of eight common peak monomer compounds of separated and preparated quaternary ammonium alkali were calculated. The results showed that coptisime, dehydrocorybulbine and columbamine for myocardial cell hypoxia/reoxygenation model of myocardial ischemia/reperfusion injury model of efficacy of activity were best.They were the important active substances.In addition, dehydrocorydaline,13-Methyldehydrocorydalmine and palmatine also had significantly protective effect. The six substances made up the active substance basis of yanhusuo against myocardial ischemia. However,13-Methylpalmatrubine and berberine had no obvious myocardial cell hypoxia/reoxygenation protective effect, which was almost the same as the multiple regression analysis, grey relational analysis and person-correlation analysis results. Therefore, the monomer dosage was a good method for verification of reliability and accuracy of fingerprint-efficacy analysis methods.In this dissertation, through the mouse hot plate method and torsion body method yanhusuo tertiary amine alkali and alkali quaternary ammonium and compatibility of both1:1were compared for myocardial cell hypoxia/reoxygenation protective effect. The results showed that, for central pain caused by heat conduction, tertiary amine alkali had obvious analgesic effect, quaternary ammonium alkali and mixed group, had no obvious analgesic action. For peripheral pain caused by intraperitoneal injection of acetic acid, alkali quaternary ammonium had obvious analgesic effects, tertiary amine alkali had no obvious analgesic action, and compatibility of them had no apparent influence of quaternary ammonium alkali for analgesic effect.Pharmacological effects were studied against myocardial ischemia on quaternary ammonium alkali, tertiary amine alkali and the tertiary amine alkali and alkali quaternary ammonium (1:1) mixed dosage groups. The experimental results showed that the group of low dose of quaternary ammonium alkali had obvious effect of anti myocardial ischemia, and anti-myocardial ischemia effect of the mixed group were better than single quaternary ammonium alkali or tertiary amine alkali. The increasing efficacy had nothing to do with dose because high doses of quaternary ammonium alkali had no significant difference with model group.The results suggested that tertiary and quaternary alkaloids might have synergistic effect against myocardial ischemia.