| Flos Lonicerae is the dried flower and bud of Lonicera japonica used as a Chinese herb in clinics.Fructus Forsythiae is one kind of traditional Chinese medicine which was officially recorded by every version of Pharmacopoeia of the People's Republic of China.It was the dried fruit of Forsythia suspensa(Thunb.) Vah1.Both of them have the bioactivities of heat-clearing and detoxicating,debugging wind-heat.The Herb Couple of Flos Lonicerae and Fructus Forsythiae is a traditional compatibility in Chinese medicine clinics.Therefore,in this dissertation,the Herb Couple of Flos Lonicerae and Fructus Forsythiae was used as a case study to find an appropriate and quick method for enclosing the effective chemical substances of the complexity in Chinese herb medicines.The core ideas of the method is as follows:on the basis of hypothesis "the constituents absorbed into blood may be the effective constituents in Chinese medicine".The correlation of the constituents absorbed into intestines in the model of rat everted gut sac and the constituents in the blood should be studied,the feasibility study of in vitro absorptive model to replace the whole animal model was done,then the fast,simple and low-cost method to find pharmacodynamic ingredients of Chinese herbs was established initially.Surrounding the core of the issue,the main pharmacodynamics functions of the Herb Couple of Flos Lonicerae and Fructus Forsythiae,the process in vivo and in vitro of the effective fraction of the Herb Couple of Flos Lonicerae and Fructus Forsythiae,the chemical compositions and the process in vivo and vitro and decoction of Flos Lonicerae,Lonicera japonica,and the Herb Couple of Flos Lonicerae and Fructus Forsythiae and the water soluble chemical constituents of the Herb Couple of Flos Lonicerae and Fructus Forsythiae were all studied.Then the fast,simple and low-cost method to find pharmacodynamic ingredients of Chinese herbs was established initially to reveal the possible pharmacodynamic ingredients of the Herb Couple of Flos Lonicerae and Fructus Forsythiae and to provide some basis of for the Herb Couple of Flos Lonicerae and Fructus Forsythiae in quality control.In pharmacodynamics aspect,the anti-inflammatory and antifebric effective of Lonicera japonica,Forsythia suspense and the different proportional Herb Couples of Flos Lonicerae and Fructus Forsythiae were studied on the basis of the experiment results of the mice's auricular inflammation model by croton oil and rat's fabricant model by yeast fungus.The result showed that the different proportional Herb Couples of Flos Lonicerae and Fructus Forsythiae are better than single Lonicera japonica and Forsythia suspense,and the proportion 1/1 of Herb Couples of Flos Lonicerae and Fructus Forsythiae is better than other proportion in anti-inflammatory and antifebric effective.After that,the anti-inflammatory and antifebric effective of Herb Couples of Flos Lonicerae and Fructus Forsythiae(1:1),fractions eluted with water and ethyl alcohol on macroporous resin were studied too by the same models, and the result showed that fraction eluted with ethyl alcohol on macro-porous resin is better than that eluted with water on macro-porous resin and itself in anti-inflammatory and antifebric effective.After getting the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae,a fast,simple and low-cost method to find pharmacodynamic ingredients of Chinese herbs is probed with the effective fraction.Main parts has been included as follows.The absorption of the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae was studied in the Caco-2 cell model,and the result showed that the relative little information is provided by the Caco-2 cell model. Then in order to obtain much more information,the absorption of the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae was also studied in the model of rat everted gut sac.The results showed that almost all the mainly chromatographic peaks could be absorbed in this model and no components were created or disappeared.After that,in order to explain the rationality directly of using the model of rat everted gut sac,the rat gastric juice and the intestinal juices samples were analyzed on HPLC-DAD after oral ministration of the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae.The findings indicated that no new main chromatographic peaks is generated or vanished in rat gastrointestinal fluid which explained the rationality directly using the model of rat everted gut sac to study the absorptive components of the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae.So as to find pharmacodynamic ingredients of Herb Couples of Flos Lonicerae and Fructus Forsythiae,the ingredients of the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae entered blood were studied.The result discovered that the main ingredients entered blood are consistent with the absorptive ingredients in the model of rat everted gut sac as the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae.Then in order to confirm further the pharmacodynamic ingredients of Herb Couples of Flos Lonicerae and Fructus Forsythiae,the internal organic distributions in rat of the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae were studied.The result showed that the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae was distributed in heart and lung,and the main ingredients in heart and lung are consistent with the absorptive ingredients in the model of rat everted gut sac as the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae.According the above results,the pharmacodynamic ingredients of Herb Couples of Flos Lonicerae and Fructus Forsythiae are the main ingredients entered blood,and the main ingredients entered blood are consistent with the absorptive ingredients in the model of rat everted gut sac as the effective fraction of the Herb Couples of Flos Lonicerae and Fructus Forsythiae.Therefore,we can educed that a fast,simple and low-cost method to find pharmacodynamic ingredients of Chinese herbs is established initially to reveal the possible pharmacodynamic ingredients of the Herb Couple of Flos Lonicerae and Fructus Forsythiae and to provide some basis of the Herb Couple of Flos Lonicerae and Fructus Forsythiae in quality control.After the fast,simple and low-cost method to find pharmacodynamic ingredients of Chinese herbs was established,the absorption of Flos Lonicerae,Fructus Forsythiae and the different proportional Herb Couples of Flos Lonicerae and Fructus Forsythiae is also studied in the model of rat everted gut sac.The findings showed that almost all the mainly chromatographic peaks could be absorbed and no components were created or disappeared which proved the fast,simple and low-cost method to find pharmacodynamic ingredients of Chinese herbs to be feasible and suggested that the proportion of pharmacodynamic ingredients can affected the pharmacodynamic effect of the couple herbs.Then the samples of Flos Lonicerae,Fructus Forsythiae and the different proportional Herb Couples of Flos Lonicerae and Fructus Forsythiae were also incubated in artificial gastric juice and artificial intestinal juice and were quantitative using HPLC.The results showed that almost all the mainly chromatographic peaks could be detected during incubation and no components were created or disappeared.It was showed that all the components in artificial gastric juice and artificial intestinal juice with different proportion compatibility in incubation could not be pertained to found any obvious changes.And no components were created or disappeared during incubating.But the contents of mainly components in artificial intestinal juice were higher than that in artificial gastric juice which indicated that artificial intestinal juice and artificial gastric juice can affect the proportion of pharmacodynamic ingredients.In order to discuss thoroughly the pharmacodynamic ingredients and concentrate the quantity of the main pharmacodynamic ingredients,the water soluble chemical constituents of the Herb Couples of Flos Lonicerae and Fructus Forsythiae were studied.Firstly,the difference in chemical constituents of Flos Lonicerae,Fructus Forsythiae and the Herb Couples of Flos Lonicerae and Fructus Forsythiae were analyzed by HPLC-DAD.The result showed that the main chromatograph peaks from the Herb Couples of Flos Lonicerae and Fructus Forsythiae can be attributed to the single Flos Lonicerae and Fructus Forsythiae,no obvious new chromatographic peaks were produced or vanished which suggested that the chemical constituents of the Herb Couples of Flos Lonicerae and Fructus Forsythiae can be studied through the simple Flos Lonicerae and Fructus Forsythiae.So the water soluble chemical constituents of Flos Lonicerae and Fructus Forsythiae were separated and purified by various chromatographic methods.16 compounds from Flos Lonicerae were obtained and identified by chemical methods and spectroscopic techniques including 6 iridoid glycosides,5 triterpenoid saponins,2 flavonoids and 3 others. 3-O-β-D-glucopyranosyl hederagenin 28-O-β-D- glucopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→6)]-β-D-glucopyranosyl ester and 3-O-β-D-glucopyranosyl-28-O-[α-L-rham-nopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→6)]-β-D-glucopyranosyl] oleanolic acid are two new compounds,2,6-dimethyl-6-hydroxyl-2,7-diene-1-octyl alcohol glucopyranoside was separated firstly from the Lonicera and sweroside, loganin and centauroside were isolated firstly from Flos Lonicerae.18 compounds from Fructus Forsythiae were obtained and 16 of them were identified by chemical methods and spectroscopic techniques including 4 lignans,3 organic acids,2 flavonoids and 6 others.Rutaecarpine was separated firstly from the Forsythia.Under the guidance of theory and clinical practice of TCM,utilizing the knowledge and technologies of phytochemistry,pharmaceutical analysis, pharmacology and pharmacokinetics,the main pharmacological effects,compatibility, chemical constituents and the process in vivo and in vitro about the Herb Couple of Flos Lonicerae and Fructus Forsythiae were studied,the possible pharmacodynamic chemical substances was discussed,research patterns of pharmacodynamic chemical substances for Chinese medicine were established initially which is a explored and innovated research for modernization of Chinese medicine. |
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