Studies On The Preparation Technology And Biological Activity Of Total Phenylethanoid Glycosides In The Genus Orobanche,and Pharmaphylogenetic Of Orobanchaceae

Objective According to the handbooks of Chinese herbal medicine and localherbs of all dynasties,many medicinal plants belonging to Orobanche are commonly used as substitutes for Cistanches Herba,such as O.coerulescens,O.pycnostachya and O.cernua,which have the traditional effects of strong tendons and strong bones,tonifying kidney and helping yang,and anti-fatigue similar to Cistanches Herba.It is found that there are lots of phenylethanoid glycosides?PhGs?in the medicinal plants of Orobanche,which have obvious pharmacological activities such as anti-oxidation,anti-aging,anti-fatigue,liver protection and immune enhancement.As a common parasitic weed among farmers in Inner Mongolia,O.cernua is rich in resources and rich in PhGs.Therefore,in the first part of this thesis,the PhGs were purified and enriched,the quality characterization method was established,and the antioxidant and hypoglycemic activities were studied.Orobanche,Cistanche and Boschniakia are grouped into Orobanchaceae,the main medicinal plants of which have phylogenetic relationship closely and similar traditional curative effects.PhGs,as representative compounds,have some advantages in both content and chemical structure diversity.They are considered to be the important pharmacodynamic material basis of these genus.Therefore,the second part of this paper is guided by the theory of phylogeny of medicinal plants,and study on,analysis and collate the characteristic chemical constituents-PhGs of Orobanche,Cistanche and Boschniakia by HPLC and UV-Vis methods.The relationship between the genus Orobanche,Cistanche and Boschniakia was discussed in order to lay a foundation for the rational utilization and comprehensive excavation of the medicinal resources of it.Methods 1.Optimization of extraction process of total PhGs by single factor and orthogonal experiment.2.The purification process of macroporous resin was determined by investigating the kinds of macroporous resin,sample concentration,flow rate,ethanol elution concentration,elution speed,volume and so on.3.Establishing the quantitative determination methods of total PhGs,acteoside,crenatoside and the extraction rate combining comprehensive evaluation method to evaluate the extraction and purification process.4.Build 12 batches of total PhGs fingerprint in Orobanche by HPLC.5.The DPPH radical scavenging activity of PhGs was determined and discussing the structure-activity relationship of antioxidant activity and PhGs.6.The diabetic model of rat was induced by streptozotocin and given different doses of total PhGs,and to study the hypoglycemic activity by observing blood glucose,glucose tolerance,insulin tolerance,serum insulin concentration and the contents of muscle and hepatic glycogen.7.Characteristic chromatogram and content determination of PhGs in Orobanche,Cistanche and Boschniakia are made by UV-Vis and HPLC.Results 1.Optimization of the preparation process of total PhGs are reasonable and feasible,the contents of PhGs,acteoside and crenatosid are reach 70%,30%and 12%,respectively.2.The HPLC fingerprint was established using 12 batches of total PhGs in Orobanche,and 15 common peaks were confirmed,9 of which were identified,the similarity of preparation at different times could reach more than 0.99.3.The DPPH radical scavenging activity of PhGs was determined by enzyme labelling apparatus,all of the total PhGs,acteoside:crenatoside=1:1 and 12 monomers showed good scavenging activity of DPPH free radical.4.Total PhGs can improve the sugar tolerance,strengthen the diabetic rats to exogenous insulin sensitivity,and can promote the secretion of pancreatic islet cells,increased serum insulin level of the diabetic rats,potential role in regulation of glucose metabolism.5.Establishment of characteristic fingerprint of PhGs in the genus Orobanche,Cistanche and Boschniakia,the results showed that the PhGs of Cistanche are more similar to those of Orobanche,and none of them similar to Boschniakia.Crenatoside and isocrenatoside are confined to the distribution of the Orobanche,and echinacoside of Cistanche.6.The contents of total PhGs are in the range of 47.77³25.43 mg·g^-1,15.46¹17.79 mg·g^-11 and56.65¹32.42 mg·g^-1,and the genus Orobanche is higher than others.Conclusion The main idea of this subject is traditional curative effect,component and biological activity.Traditional pharmacology,phytochemistry,analytical chemistry,pharmacology and mathematical statistics were used in the study to obtain high purity PhGs,the quality can be effectively characterized by HPLC fingerprint,and the research on hypoglycemic activity has also been explored and tried.The characteristic fingerprint and the determination method of PhGs in the genus Orobanche,Cistanche and Boschniakia are to reflect the differences between genus and within genus.