| The plants in Schisandraceae includes two genus such as Schisandra Michx and Kadsura Kaempf.ex.Juss,in which there are more than 60 species in the world.In our country,there are about 29 species found in Schisandraceae.So far,the fruits of Schisandra chinensis(Turcz.)Baill was included in "Chinese Pharmacopoeia"(2015 version)as North WUWEIZI.In addition,the fruits of Schisandra sphenanthera Rehd.Et Wils are also included in "Chinese Pharmacopoeia"(2015 version)as Sorthern WUWEIZI.They are the legal drug source plants of WUWEIZI.Previous reports have showed that the plants in Schisandraceae exerted a wide range of pharmacological activities,especially for the investigation on active compotents including lignans and triterpenoids.The findngs from previous results confirmed that the content of lignans was greatly different in different varieties of WUWEIZI.For example,the content of schisandrin in S.chinensis should not be less than 0.4% and the content of schisantherin A in S.sphenanthera should not be less than 3.0 % in "Chinese Pharmacopoeia"(2015 version),respectively.The content of deoxyschizandrin and schisantherin A in S.sphenanthera were higher than that in S.chinensis,but the contents of schisandrol A,schisandrol B,?-Schizandrin and total lignans in S.sphenanthera were relatively lower than S.chinensis.Thus,the huge difference between the species of WUWEIZI plants from the external form to the internal composition of the compounds are showed clearly.The distant grafting is one of the technical means to realize the moving north of the southern species,and provides the basic conditions for the study of the Kadsura genus in different habitats.In this thesis,the effects of distant grafting on physiological and biochemical characteristics of S.chinensis and K.longipedunculata were investigated for the first time.In addition,the effects of distant grafting on the photosynthetic system and the synthesis pathway of lignans were evaluated using proteomics techniques.Importantly,the pharmacological analysis of S.chinensis(WWZ)in urine on type 2 diabetic rats was established in the presnt study.The results in this thesis were summersized as follows:(1)The chlorophyll fluorescence parameters of S.chinensis was analyzed by the modulation of chlorophyll fluorescence imaging system IMAGING-PAM.The results showed that the ?PS?,qL and rETR of scion increased significantly after grafting.Nutrients in S.chinensis could be transmitted to K.longipedunculata and be used for effective photosynthesis,and the accumulation of organic matter of K.longipedunculata was also accelerated.(2)The hormone content of K.longipedunculata were analyzed after distinct grafting.Results clear showed that the contents of IAA,GA,ZR,and the ratio of ZR/IAA and ZR/ABA in the scion of K.longipedunculata were increased by the grafting,and the content of ABA was decreased.The increase of IAA and GA and the decline of ABA clearly indicates that the grafting is beneficial to the vegetative growth of K.longipedunculata,and its growth potential is stronger than that of ungrafted seedlings.The increase of ZR/IAA showed that the growth of lateral buds of grafted seedlings was not inhibited.The increase of ZR/ABA indicated the better plant growth.So grafting increased the contents of vegetative growth related hormones and enhance the adaptability of K.longipedunculata in the northeast of China.(3)Through the differential protein pathway analysis,we found one photosynthetic metabolic pathway related to grafting.Interestingly,the different proteins and the genes on this pathway were all upregulated after the grafting,so that the photosynthetic performance of grafted seedlings is stronger than that of ungrafted seedlings,and the growth potential is enhanced.This was consistent with the results of the increased actual photosynthetic efficiency and photosynthetic electron transport rate of K.longipedunculata caused by grafting,which is consistent with the results of the graft growth rate indicated by the change of endogenous hormone content.(4)By using the iTRAQ proteome analysis techenology,the upstream regulatory genes and proteins related to the synthesis of Schisandra lignans were found.These regulatory enzymes(beta-glucosidase,cinnamyl-alcohol dehydrogenase CAD,peroxidase and coniferyl-aldehyde dehydrogenase)were down-regulated after grafting and could provide more raw materials for synthetic lignans.The synthetic downstream genes of lignans are yet to be further excavated and validated.(5)In this study,an untargeted urinary metabonomics method based on UPLC-Q-TOF-MS has been established to investigate the mechanism of Schisandra chinensis in treating diabetes and its complications.The urinary biomarkers related to the therapeutic effects of Schisandra chinensis on the diabetes rats were analyzed.In urine,28 endogenous metabolites were identified as potential biomarkers,including 13 in positive ion mode,16 in negative ion mode and and hippuric acid detected both in positive and negative ion modes.The results revealed that Schisandra chinensis mainly affected the pathways of pentose and glucuronate interconversions,riboflavin metabolism,pantothenate and CoA biosynthesis,arginine and proline metabolism,intestinal bacteria metabolism,ascorbate and aldarate metabolism and tryptophan metabolism in diabetic rats.Combined with biological analysis of these pathways,the therapeutic mechanism of Schisandra chinensis on diabetes and its complications was verified.Based on the biological function of each pathway,the effect of Schisandra chinensis on diabetic nephropathy is stronger.Moreover,it also has the effects of protecting liver,decreasing fat and antioxidant activity. |
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