| Rationales:Radix Polygalae, the root of Polygala tenuifolia Willd or Polygala sibirica L., a traditional Chinese herb has long been used as a traditional Chinese medicine and widely used in the vast majority of Chinese medicine prescription for antidepressant, expectorant, antitussive and anti-inflammatory et al. At present, the studies of Polygala tenuifolia Willd mainly concentrated in①the aerial part of P. tenuifolia is discarded usually after the root harvest, resulting in waste of resources.②the xylem is required to be removed traditionally, not required now.③the processed products and so on.Quality evaluation of P. tenuifolia still have the following deficiencies:①quality evaluation of P. tenuifolia by limited ingredients to cannot do a comprehensive analysi of the chemical composition, and HPLC-UV fingerprints only can do simple chromatographic analysis. It is very important to introduce a more comprehensive analytical tool-metabolomics;②he acid hydrolyzates of saponins cannot be used as reference substance to evaluate saponins content comprehensively. Alkaline hydrolyzates of saponins is single, Chinese Pharmacopoeia2010edition provides tenuifolin to saponin content measured by the standard, there is no report of saponin content evaluation by tenuifloin;③the aerial part of P. tenuifolia is discarded usually after the root harvest, resulting in waste of resources. The xylem is required to be removed traditionally, not required now. And the evalution of different parts of P. tenuifolia only concentrated in a limited composition;④pharmacological studies of the aerial part have not been reported.Objective:the methods based on NMR/UPLC plant metabolomics research and content of determination of tenuifolin by UPLC have been established. Then the metabolomic profiles of different parts and different processed products of P.tenuifolia have not be evaluated, and the pharmacological efficacies of the aerial part of P. tenuifolia have been studied, providing the scientific guidance from new view for the quality assessment and resource utilization of P. tenuifolia.Methods:1. The study based on NMR and UPLC plant metabolomics of the root, aerial part, cortex, xylem, Honey-stir-baked Radix Polygalae, Liquorice juice-stir-boiled Radix Polygalae:First, the extracts of samples were determinated by NMR spectrometer, the1H NMR spectras were processed by MestRenova software for large amounts of data, which was analysised for data principal component analysis and the smallest partial squares analysis by SIMC A-P software. Then, part of the metabolites in the NMR spectrum and different metabolites in samples were identified according to the literature and study results, combining with the t-test, the relative concentration of the different metabolites in the root, aerial part, cortex, xylem, H P. tenuifolia and L P. tenuifolia were determinate, to seek for NMR metabolomics differences among the different medicinal parts and processed products of Polygala tenuifolia Willd.2. The study based on UPLC plant metabolomics of the root, aerial part, cortex, xylem, H P. tenuifolia and L P. tenuifolia:First, the extracts of samples were determinate by Ultra High Pressure Liquid Chromatography (UPLC) fingerprint, the peak areas in the chromatograms were integrated manually. Then the datas were analysised for data principal component analysis and the smallest partial squares analysis by SIMCA-P software. Then, part of the metabolites in the UPLC chromatogram and different metabolites in samples were identified according to the literature and study results, combining with the t-test, the relative concentration of the different metabolites in the root, aerial part, cortex, xylem, H P. tenuifolia and L P. tenuifolia were determinated, to seek for UPLC metabolomics differences among the different medicinal parts and processed products of Polygala tenuifolia Willd.3. The UPLC content determination of tenuifolin in the root, aerial part, cortex, xylem, H P. tenuifolia and L P. tenuifolia:The standard substance for UPLC content determination, tenuifolin, was separated by us in the laboratory, the purity more than98%.4. The antidepressant and antitussive, expectorant effects of the aerial part of Polygala tenuifolia Willd were studied by the classic Mice tail suspension test and Forced swimming test antidepressant model, the classic ammonia induced cough test and racheal phenol red excretion test antitussive, expectorant model.Results:1. NMR plant metabolomics studies have shown that, there were significant differences among the metabolomic profiles of different parts and processed products, which mainly concentrated in the primary metabolites.①the metabolomes of different parts of Polygala tenuifolia Willd were different. Comparing with the root, the contents of valine, glutamic acid, succinic acid, GAB A, tartaric acid, maleic acid and sinapic acid were higher in the aerial part, and the contents of threonine, alanine, aspartic acid, acetic acid, sucrose, adenine and saponins compounds were lower. Compared with the phloem, the contents of alanine was higher in the xylem, the contents of threonine, aspartic acid, acetic acid, succinic acid, GAB A, tartaric acid, formic acid, choline, fructose and sucrose were lower;②the metabolomes of different processed products of Polygala tenuifolia Willd were different, too. Honey-stir-boiled made the contents of formic acid, and α-glucose increase, the contents of threonine, alanine, fructose and sucrose decreased; Liquorice juice-stir-boiled made the contents of leucine, tartaric acid increased, threonine, alanine, butanedioic acid, fumarate and sucrose decrease; Meanwhile, comparing with H P. tenuifolia, the contents of formic acid, and α-glucose were lower in L P. tenuifolia.2. UPLC plant metabolomics studies have shown that, there were significant differences among the metabolomic profiles of differert parts and processed products, which mainly concentrated in the secondary metabolites.①the metabolomes of different parts of Polygala tenuifolia Willd, were different. Comparing with the roots, the content of Polygalaxanthone III was higher in the aerial parts, and the contents of3,6'-disinapoylsucrose ester, Sibiricose A5, Sibiricose A6, Tenuifoliside A, Tenuifolise I, Tenuifolise H, PolygalaxanthoneⅣ, Onjisaponin A and Onjisaponin K were lower. Compared with the phloem, the contents of Tenuifolise I was higher in the xylem, the contents of Tenuifoliside A, Tenuifolise I, Tenuifolise H, Onjisaponin A, Polagalasaponin X X X II were lower;②the metabolomes of different processed products of Polygala tenuifolia Willd were different, too. Honey-stir-boiled made the contents of Tenuifolise I Onjisaponin A increase, the contents of3,6'-disinapoylsucrose ester, Tenuifoliside A, Tenuifolise H, Polygalaxanthone IV decrease; Liquorice juice-stir-boiled made the contents of Tenuifolise I, Tenuifolise H, Onjisaponin K increased,3,6'-disinapoylsucrose ester, Polygalaxanthone IV, Tenuifolise H decreased; Meanwhile, comparing with H P. Tenuifolia., the contents of3,6'-disinapoylsucrose ester and Tenuifolise I were lower in L P. Tenuifolia.3. UPLC content determinations have shown that,①the content of tenuifolin in the aerial part was higher than the root; the xylem which required to remove, the content of tenuifolin was very lower than in cortex, and the content difference was very significant;②In traditionally Preliminary processing of P. Tenuifolia., Honey-stir-boiled made the content of tenuifolin lower, Liquorice juice-stir-boiled made the content higher, and the later the content difference was very significant.4. Pharmacological studies of the aerial part of P. Tenuifolia. have shown that, the aerial part had low antidepressant activity and strong antitussive, expectorant activities; Comparing with the roots of P. Tenuifolia. and the classic antitussive, expectorant drug carbetapentane and T. farfara, the antitussive effect of the aerial part was stronger, the expectorant effect was lower.Conclusion:In the paper, the NMR and UPLC plant metabolomics and UPLC content determination of tenuifolin in P. tenuifolia were established, at the same time, the antidepressant and antitussive, expectorant activities of the aerial parts were evaluated. The results showed that the metabolomic profils of the root, aerial part, cortex, xylem, Honey-stir-baked Radix Polygalae, Liquorice juice-stir-boiled Radix Polygalae had significantly differences, and the aerial parts have certain antitussive and expectorant activity, and antitussive acticity is stronger than the root, expectorant activity is weaker. The active compound might be Polygalaxanthone III. |
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