Chemical Characterization Of Aqueous Extracts Of Polygala Tenuifolia And Study On The Biological Effect Of "MEN"

Rationales: Polygala tenuifolia is one of the major medicinal herbs in China,with the effects of tranquilizing the mind,educating the mind and kidneys,dispelling phlegm,and reducing swelling.The traditional medicinal part of P.tenuifolia is the root bark of P.tenuifolia,which needs to be extracted from the xylem during use.At present,except for the special grade root bark,the other grades have different degrees of adulteration of xylem,and the extraction of xylem causes waste of resources and also has a certain impact on the environment.The heart removal of P.tenuifolia is mainly related to the record of Lei Gong Pao Zhi Lun that "before using P.tenuifolia,the heart needs to be removed;if the heart is not removed,it will make people " Men " when taking it".It is necessary to clarify the connotation of "Men",and it is necessary for the research related to the removing of the xylem of P.tenuifolia.Objective: This study was conducted to investigate the differences between the xylem and the root bark of P.tenuifolia based on metabolomics,to investigate the possible biological effects of the xylem of P.tenuifolia on "Men" using behavioral studies,and to elucidate the meaning of P.tenuifolia at the molecular level using network pharmacology techniques.Methods:(1)We used Nuclear Magnetic Resonance(NMR)and Ultra Performance Liquid Chromatography-Quadrupole-Time of Flight Mass Spectrometry(UPLC-Q TOF-MS)to collect data from the xylem and root bark samples of P.tenuifolia,and used various strategies including molecular network techniques to resolve the chemical composition of the xylem(PTX)and root bark of P.tenuifolia(PTRB).We used non-targeted plant metabolomics techniques to resolve the differences in chemical composition between the root bark and xylem of Cosmopolitan.(2)We used sucrose preference test 、 forced-swimming test 、 open field test 、elevated-plus maze test 、 Morris water maze test to reveal the behavioral effects of aqueous extracts of PTX on SD rats after gavage,and to reveal the biological effects of "Men".(3)Using intracerebral cannula injection,we administered phencyclidine N-methyl-D-aspartate(NMDA)receptor antagonist 2-amino-5-phosphonopentanoic acid(AP5)to the lateral receptor nucleus(LHb)of rats gavaged with aqueous extract of PTX,and verified whether the behavioral effects of aqueous extract of PTX on SD rats were related to the lateral receptor nucleus and NMDA receptors.(4)Network pharmacology techniques,combined with non-targeted plant metabolomics screening of differential metabolites up-regulated in the PTX relative to the PTRB,were used to predict the core targets and key pathways of PTX affecting depression-like and anxiety-like behaviors,and We also use molecular docking technology to validate the screened core components and core targets of the PTX that cause "MEN"..Results:(1)A total of 25 chemical components were identified using NMR;137 components were identified using UPLC-Q TOF MS,including 94 components in negative ion mode and 43 components in positive ion mode.12 differential metabolites were screened by NMR;36 differential metabolites were screened by UPLC-Q TOF MS in negative ion mode,and 43 differential metabolites were screened by positive ion NMR screened a total of 12 different metabolites;UPLC-Q TOF MS screened 36 different metabolites in negative mode and 13 in positive mode.(2)The behavioral effects of the aqueous extract of the PTX on SD rats after gavage were investigated in multiple ways using the SPT,FST,OFT,EPM and MWM,revealing that the occurrence of "Men" is dose-dependent and transient,and its objective manifestation may be a depression-like and anxiety-like behavior.(3)The mechanism of depression phenotype generation was used to verify the possibility of "Men" as a depression-like behavior.The results showed that cannula injection of AP5 into the LHb brain region reversed the results of depression-like behavioral test in rats induced by aqueous extracts of PTX,but did not significantly affect the results of anxiety-like behavioral test.The behavioral effects of aqueous extracts of Xylem of P.tenuifolia in SD rats were related to the lateral rein nucleus and NMDAR.(4)Using network pharmacology techniques combined with non-targeted phytometabolomics screening of differential metabolites up regulated in the PTX relative to P.tenuifolia root bark,nine potential components were identified that may act through91 potential targets including SLC6A1,GRM5,GRM8,GRIA1,and SLC1A1,involving neuroactive ligand-receptor interactions,retrograde endogenous cannabinoid signaling,the serotonergic synapses,c AMP signaling pathway,calcium signaling pathway,and other possible pathways to affect the release of dopamine and other neurotransmitters and other pathways to influence the production of depression-like and anxiety-like behaviors in humans;and the core components and targets were validated using molecular docking techniques.Conclusion: In this study,we resolved the chemical components in the PTX and PTRB,identified 137 components,and screened the differential metabolites in the PTX and PTRB under different collection techniques,respectively,of which 12 were screened by NMR,36 by UPLC-Q TOF MS in negative,and 13 by UPLC-Q TOF MS in positive ion mode.The behavioral effects of aqueous extracts of PTX on SD rats after gavage were investigated by animal experiments,which revealed that "Men" may be a depressive and anxiety-like behavior.It was verified that the depression-like behavior was reversed by the NMDA receptor antagonist AP5,suggesting that the behavioral effects of the aqueous extract of the PTX on SD rats were related to the lateral rein and NMDA receptors.The network pharmacology results showed that some amino acids and small organic acids in the PTX could act on multiple targets and affect multiple pathways to achieve the "Men" effect.The present study opens a new perspective for the research related to the "Removing the PTX" and lays the foundation for the quality control research of the herb.