| Gegen Qinlian decoction(GGQLD) originated from Treatise on Febrile Diseases by Zhang Zhongjing, composed by Puerariae Radix (roots of Pueraria lobata (Willd.) Ohwi, PR), Scutellariae Radix (roots of Scutellaria baicalensis Georgi SR), Coptidis Rhizoma (rhizomes of Coptis chinensis Franch, CR) and honey-processed Glycyrrhizae Radix (roots of Glycyrrhiza uralensis Fisch, GR), and used for treatment of damp-heat and diarrheain common. Now, it is used in the treatment of diabetes Spleen Damp, and its clinical effectiveness has been confirmed by pharmacodynamics experiments. But the action mechanism of GGQLD and the transport, metabolism of its active ingredients is still unclear. The study hotspot is to clarify the effect of compound Chinese medicine for the disease in traditional Chinese medicines (TCM). Till now, the mechanism of GGQLD on complex diseases like type 2 diabetes and ulcerative colitis is not clear, the studies in vivo absorption and metabolism of GGQLD need to be clear. In recent years, liquid chromatography-mass spectrometry (LC-MS) is an important tool in study of metabonomics and pharmacokinetic for traditional Chinese medicine. In this thesis, LC-MS combined with metabonomics and pharmacokinetic study methods was used for the study of GGQLD to explore metabolic pathway of GGQLD treatment of type 2 diabetes and ulcerative colitis in rats. The in vivo metabolism and intestinal absorption of active ingredient were also studied, which could prove the scientificalness of the clinical efficacy and provide reference for the clinical regimen.The core of the research paper was to establish the metabolic pathway analysis of type 2 diabetes and ulcerative colitis after GGQLD treatment based on UHPLC-Q-TOF/MS. Also, the methods for the research of the in vivo metabolic rule of 11 active ingredients in rat plasma after orally administrated GGQLD and the intestinal absorption rule of GGQLD active ingredients in rat everted sac model based on HPLC/MS/MS were developed. The main research contents are as follow:1. Taking active ingredients puerarin, baicalein, baicalin, wogonoside, wogonin, liquiritin, berberine, palmatine in GGQLD as analytes, a method using multi-wavelength reverse phase high performance liquid chromatography (MWD-RP-HPLC) for the simultaneous determination of these active ingredients in GGQLD was established. This study provided quality control evaluation of GGQLD for the subsequent study on its pharmacokinetics, intestinal absorption, and metabonomics of the GGQLD treatment of STZ-induced diabetic rats and rats with ulcerative colitis.2. The mtabonomics study onstreptozocin(STZ)-induced type 2 diabetes rat after GGQLD treatment was performed using UHPLC-Q-TOF/MS techniques, as the model.The results confirmed GGQLD treatment of type 2 diabetes effect from the overall metabolic regulation mode and identified seven potential biomarkers mainly related to metabolic pathways such as sphingosine metabolic, CoA biosynthesis, primary bile acid biosynthesis and niacin synthesis. In this study, we found that GGQLD plays a therapeutic role in type 2 diabetes by regulating these metabolic pathways imbalance.3. UHPLC-Q-TOF/MS was used to analyze the variation of component in ulcerative colitisrats plasma and pattern recognition after GGQLD treatment. It was also used to find and explain potential biomarkers, match related metabolic pathways and metabolic networks. At last, the mechanism of therapeutic effect of GGQLD on ulcerative colitis was elaborated.4. The accurate and sensitive HPLC-MS/MS method was developed to determinate 11 active ingredientsin rat plasma simultaneously, includingpuerarin, daidzein, baicalin, baicalein, wogonoside, wogonin, liquiritin, liquiritigenin, berberine, jatrorrhizine andpalmatine.The proposed method was used to study the pharmacokinetic characteristics of the 11 active ingredients in rat plasma after orally administrated GGQLD. The results elaborated pharmacokinetic interactions between pharmacology and GGQLD effective in a variety of biologically active ingredients.5. The accurate and sensitive HPLC-MS/MS method was developed todeterminate 11 active ingredients in rat intestinal absorption liquid simultaneously, includingpuerarin, daidzein, baicalin, baicalein, wogonoside, wogonin, liquiritin, liquiritigenin, berberine, jatrorrhizine andpalmatine. The proposed method was used to study absorption kinetics of 11 active ingredients of GGQLD in rat everted gut sac model and compare the difference between flavonoids and alkaloids in each component absorption effect, in order to select the appropriate markers to characterize the pharmacokinetics makers and provide the basis for pharmacokinetic characterization of the entire prescription. |
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