Study On Antitussive And Expectorant Substances Of Total Saponins Of Platycodonis Radix And Their Mechanism Based On Spectrum Effect Relationship,Microbiota Transform And Metabonomics

Platycodins Radix(PR)is the dried root of P.grandiflorum(Jacq.)A.DC.It possessed the functions of promoting lung,soothing pharynx,expectorant and purging pus.The main active fraction of PR is total saponins(PTS).However,the active components and their mechanisms of PTS about antitussive and expectorant are not clear.On the other hand,the oral administrated Chinese herbal medicines could interact with the intestinal flora when they entered the intestinal tract.During this process,the structure,activity,and mechanism of the components may be changed significantly.The spectrum-effect relationship(SER)combined "spectrum" with "effect",which can be used to quickly screen the potential active ingredients.While the metabolomics can be applied to reveal the mechanism of action of drugs.Based on those,the active components of the PTS with antitussive and expectorant activities were clarified using SER;and their mechanism were studied using metabonomics.Furthermore,the PTS were biotransformed by intestinal bacteria and the products were enriched for the study of the active metabolites with antitussive and expectorant effect were also studied using SER.And their mechanism was also studied using metabonomics method.The main methods and results were listed as follows:1.The PTS fingerprints of 10 PR samples from different origins were constructed using UHPLC-LTQ-Orbitrap-MS technology.The antitussive and expectorant effects of PTS from 10 different origins were evaluated by the concentrated ammonia water spray method in mice and excretion of phenol red in the trachea the method,respectively.The SER between antitussive and expectorant activities of PTS and fingerprint spectrum was constructed by partial least squares regression analysis(PLS),respectively.The metabolomics was applied to explore the antitussive and expectorant mechanism of PTS.The results of the SER study showed that Polygalacin D、Deapioplatycodin D、Platycodigenin、Platycodin D、Platycoside G3、3"-O-acetyl platycodin D3 、 Platycodin D3 、 β-Gentiotriosyl-platycodigenin 、 3-O-β-D-glucopyranosyl platycodigenin)and 2"-O-acetyl polygalacin D are related to the antitussive activity of PTS,while 3"-O-acetyl platycodin D3,Platycodin D2,Platycoside D,Polygalacin D2,Tangshenoside I,Platycodin H,Deapio-platycodin D3,Conyzasaponin T,Platycoside E,Deapio-platycoside E,Platycodon B,Platycodin D,Platycoside G3,Platycogenin A,Platycoside G2,and Platycodin D3 are related to the expectorant activity of PTS.Linoleic acid metabolism,arachidonic acid metabolism and glycerophospholipid metabolism were the potential metabolism pathways related to antitussive activity.While linoleic acid metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,arachidonic acid metabolism,phenylalanine metabolism and α-linolenic acid metabolism were related to the expectorant activity.2.The PTS of 10 PR samples were biotransformed in vitro by intestinal bacteria.UHPLC-LTQ-Orbitrap-MS technology was used to construct the fingerprint of PTS intestinal bacteria biotransformed product(PTS-IBTP).The antitussive and expectorant effects of the 10 PTS-IBTP were evaluated by the concentrated ammonia spray method and excretion of phenol red in the trachea the method,respectively.PLS was used to construct the SER between the antitussive and expectorant activity of the PTS-IBTP,respectively.The metabolomics was applied to explore the antitussive and expectorant mechanism of PTS-IBTP.The results of antitussive SER showed that IBTP-18、IBTP2、IBTP-4、IBTP-13、IBTP-6、IBTP-17、IBTP-11、IBTP-16、IBTP-22、IBTP-20 and IBTP-21 were related to antitussive activity.The results of expectorant SER showed that IBTP-11、IBTP-22、IBTP-19、IBTP-18、IBTP-17、IBTP-23、IBTP-13、IBTP-5、IBTP-21、IBTP-10、IBTP-2 and IBTP-16 were related to expectorant activity.The results of metabolomics showed that PTS-IBTP could exert antitussive activity through linoleic acid metabolism,arachidonic acid metabolism and glycerolphospholipid metabolism.While linoleic acid metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,arachidonic acid metabolism,phenylalanine metabolism and α-linolenic acid metabolism were related to the expectorant activity.3、The effective forms or the main effective forms of PTS in the body may be the original and non-prototype components biotransformed by the intestinal bacteria.In addition,many components in PTS many possess the same or similar effects as they have the same parent structure,which could be transformed by intestinal bacteria to form the metabolites with the same or similar structure in the body.Hence,many compounds in PTS may have similar biological activities and similar metabolomics regulation mechanisms.