Studies On The Chemical Components And Uric Acid Lowering Activity Of 5 Ethnic Medicinal Plants From Genus Clematis

The genus Clematis of Ranunculaceae,including 300 species,is distributed in all over world.There are more than 150 species in China,most of which are widely distributed in southwest China with abundant resources.Forty nine plants from Clematis were considered to be the ethnic medicines,possessing antibacterial and anti-inflammatory,inducing diuresis,dispelling wind and relieving pain,in 29minorities throughout China.Some ethnic medicines were used as the treatment for palpitations,ruffians,stomach cold,abdominal distension,nasosinustis,jaundice,and so on.There are some differences,which showed the diversity of its efficacy and provided the valuable information for the development of the plants from Clematis,in medicinal species,location and clinical application among different nationalities.However,the efficacy and scientific connotation of these herb medicines need to be verified and clarified.On the other hand,synonym,homonym,and succedaneum are presented very seriously due to the distribution of these herb medicines.But these phenomena have never been interpreted through science thought and science method.Thus,it is important to rationally exploit and utilize resources,and to standardize clinical application of the genus Clematis that the chemical constituents,pharmacological activities and action mechanism between the different species of Clematis or the same species in different nationalities are compared.In this thesis,the chemical constituents of five plants of the genus Clematis were investigated;the activities of reducing uric acid in vitro or vivo of the pure compounds were evaluated.The main contents and results are as follows:1 The triterpenoid saponins of C.rehderiana,C.akebioide,C.speterae,C. songarica,and C.delavayi were analyzed with LTQ-Oribitrap XL Mass on basis of the fragmentation regularity of triterpenoid saponins isolated from the plants of the genus Clematis.Thirty eight triterpenoid saponins of C.rehderiana,including 21hederagenin-type triterpenoid saponins,were identified and the exact structures of nine compounds were confirmed.Fouty six triterpenoid saponins of C.akebioides,including 40 hederagenin-type triterpenoid saponins,were identified and the exact structures of ten compounds were confirmed.Twety one triterpenoid saponins of C.songarica,including 10 hederagenin-type triterpenoid saponins,were identified and the exact structures of seven compounds were confirmed.Four triterpenoid saponins of C.peterae,including 3 Oleanolic-type triterpenoid saponins,were identified and the exact structure of one compound was confirmed.Twety four triterpenoid saponins of C.delavayi,including 17 hederagenin-type triterpenoid saponins,were identified and the exact structures of four compounds were confirmed.2 Six unknown compounds,including one coumarin?24?,one lignin?25?,and four hederagenin-type triterpenoid saponins?36,37,93,94?,along with seventy two known compounds was isolated from five plants by various chromatography methods?silica gel column chromatography,MCI,ODS column chromatography,Sephadex LH-20 column chromatography,MPLC,RPLC?.Their structures were elucidated on the basis of ¹H NMR,¹³C NMR,¹H-¹H COSY?HSQC?HMBC?MS.Thirty compounds were obtained for the first time.Thirteen compounds were the common components among 2 or more plants,indicating that there were significant differences in structure types and composition of the isolated compounds among 5 species of Clematis.3 The uric acid lowering activities and mechanism of five compounds from two or more plants and five new compounds were assayed with the inhibitory activity of XOD and HK-2 cell.The activity reducing uric acid of calcoside D,a gram-level high-content compound obtained from all five plants,was validated with animal model.The results showed ten compounds had little inhibitory effect on XOD in vitro,but affected the expression of three uric acid transporters in different levels,indicating potential the effect reducing acid.Calcoside D can significantly reduce the level of uric acid and urea nitrogen in serum of the model,suggesting that it has a certain role in reducing uric acid and protecting kidney.Calcoside D could decrease XOD activity in the liver of the mice at high and low doses,but the effect of calcoside D was not as good as that of the allopurinol group.Comparing with the model group,OAT1expression was increased and URAT1 and GLUT9 protein expression was decreased in the calcoside D group.Meanwhile,the selectivity of calcoside D was higher than that of benzbromarone.These results suggested that calcoside D may reduce uric acid level by inhibiting XOD activity,inhibiting the reabsorption of uric acid,and promoting the transport of uric acid,and had fewer side effects than benzbromarone.Conclusion:Those results supported why five plants possess the varied clinical efficacy among different nationalities and are often usedalternately,provided chemical and preliminary pharmacodynamic evidence for the rational utilization of resources and variety systematization of five plants,and offer the reason for that C.rehderiana Craib?C.akebioides?Maxim.?Hort.ex Veitch?C.delavayi Franch were use as“Ye meng”medicinal materials.However,more studies of philology,resources,chemistry,and pharmacology were needed to support the rational usage of those plants because“Ye meng”medicinal materials were used for long time in the Zang nationality.