A strategy combining a tailored database and high-throughput activity screening that discover bioactive metabolites derived from Magnoliae Officinalis Cortex(MOC)was developed and implemented to rapidly profile and di...A strategy combining a tailored database and high-throughput activity screening that discover bioactive metabolites derived from Magnoliae Officinalis Cortex(MOC)was developed and implemented to rapidly profile and discover bioactive metabolites in vivo derived from traditional Chinese medicine(TCM).The strategy possessed four characteristics:1)The tailored database consisted of metabolites derived from big data-originated reference compound,metabolites predicted in silico,and MOC chemical profile-based pseudomolecular ions.2)When profiling MOC-derived metabolites in vivo,attentions were paid not only to prototypes of MOC compounds and metabolites directly derived from MOC compounds,as reported by most papers,but also to isomerized metabolites and the degradation products of MOC compounds as well as their derived metabolites.3)Metabolite traceability was performed,especially to distinguish isomeric prototypes-derived metabolites,prototypes of MOC compounds as well as phase I metabolites derived from other MOC compounds.4)Molecular docking was utilized for high-throughput activity screening and molecular dynamic simulation as well as zebrafish model were used for verification.Using this strategy,134 metabolites were swiftly characterized after the oral administration of MOC to rats,and several metabolites were reported for the first time.Furthermore,17 potential active metabolites were discovered by targeting the motilin,dopamine D2,and the serotonin type 4(5-HT4)receptors,and part bioactivities were verified using molecular dynamic simulation and a zebrafish constipation model.This study extends the application of mass spectrometry(MS)to rapidly profile TCM-derived metabolites in vivo,which will help pharmacologists rapidly discover potent metabolites from a complex matrix.展开更多
OBJECTIVE:To eliminate ineffective or interfering compounds from the eight absorbed compounds(8ACs),identify the primary ACs that represent the multifunctional therapeutic effects of Zhiqiao(Fructus Aurantii Submaturu...OBJECTIVE:To eliminate ineffective or interfering compounds from the eight absorbed compounds(8ACs),identify the primary ACs that represent the multifunctional therapeutic effects of Zhiqiao(Fructus Aurantii Submaturus)and Houpo(Cortex Magnoliae Officinalis)(FM),along with elucidating their underlying mechanisms.METHODS:Key multifunctional ACs were screened through ex vivo-to-in vitro extrapolation(ex vivo dose=serum concentration)and validated in vivo,with efficacy assessed via contribution(dose=FM content).Functional magnetic resonance imaging analyzed brain regions's blood oxygen level-dependent(BOLD)changes,and the molecular mechanisms were analyzed by transcriptome of the dentate gyrus(DG).RESULTS:The results showed that representative 2ACs(Nobiletin+Magnolol)and 3ACs(Nobiletin+Magnolol+Meranzin hydrate)screened on ex vivo experiment by the criteria of contribution ranking contributed 80.72%-126.74%of the antidepressant and prokinetic effects of the FM(improvement of depressive-like behaviours,gastrointestinal disorder,monoamine neurotransmitters,ghrelin,endocrine hormones,pro-inflammatory factors,oxidative stress indicators).In addition,3ACs demonstrated superiority over 2ACs in improving depression and levels of multiple stress mediators.Zhiqiao(Fructus Aurantii Submaturus)(FRA)reduced acute stress-induced hyperactivation of the cingulate cortex,thalamus,hypothalamus,and entorhinal cortex and elevated BOLD signaling in the insular cortex,temporal association cortex.Furthermore,FRA upregulated pathways of neurotransmitter receptor activity and serotonergic synaptic function and downregulated inflammation-related pathways at the genetic level.CONCLUSION:2ACs and 3ACs closely reflected FM's multifunctional effect in antidepressant and prokinetic.FRA restores stress-impaired neural connectivity in functional brain regions and improves synaptic plasticity and neurogenesis at the genetic level.展开更多
A novel high performance liquid chromatographic method was developed for the determination of 4-O- methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation. Plasma samples were treated by o...A novel high performance liquid chromatographic method was developed for the determination of 4-O- methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation. Plasma samples were treated by one-fold volume of methanol and acetonitrile to remove the interference proteins. A reverse phase column of SHIM- PACK VP-ODS （150 mm × 4.6 mm, 5.0 μm） was used to separate 4-O-methylhonokiol in the plasma samples. The detection limit of 4-O-methylhonokiol was 0.2 μg/L and the linear range was 0. 012 - 1. 536 mg/L. The good extraction recoveries were obtained for the spiked samples （84.7%, 89.3% and 87.7% for low, middle and high concentrations of added standards, respectively）. The relative standard deviation of intra-day and inter-day precisions was in the range from 0.6% to 13.5%. The pharmacokinetic study of 4-O-methylhonokiol was made and the results from the plasmaconcentration curve of 4-0-methylhonokiol showed a two-apartment open model. This work developed a sensitive, stable and rapid HPLC method for the determination of 4-O-methylhonokiol and the developed method has been successfully applied to a pharmacokinetic study of 4-O-methylhonokiol.展开更多
基金supported by the Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences,China(Grant Nos.:CI2023E002 and CI2021A04513)the National Natural Science Foundation of China(Grant Nos.:82204619 and 82274094)the Fundamental Research Funds for the Central Public Welfare Research Institutes,China(Grant Nos.:ZZ15-YQ-067 and ZZ16-ND-10-26).
文摘A strategy combining a tailored database and high-throughput activity screening that discover bioactive metabolites derived from Magnoliae Officinalis Cortex(MOC)was developed and implemented to rapidly profile and discover bioactive metabolites in vivo derived from traditional Chinese medicine(TCM).The strategy possessed four characteristics:1)The tailored database consisted of metabolites derived from big data-originated reference compound,metabolites predicted in silico,and MOC chemical profile-based pseudomolecular ions.2)When profiling MOC-derived metabolites in vivo,attentions were paid not only to prototypes of MOC compounds and metabolites directly derived from MOC compounds,as reported by most papers,but also to isomerized metabolites and the degradation products of MOC compounds as well as their derived metabolites.3)Metabolite traceability was performed,especially to distinguish isomeric prototypes-derived metabolites,prototypes of MOC compounds as well as phase I metabolites derived from other MOC compounds.4)Molecular docking was utilized for high-throughput activity screening and molecular dynamic simulation as well as zebrafish model were used for verification.Using this strategy,134 metabolites were swiftly characterized after the oral administration of MOC to rats,and several metabolites were reported for the first time.Furthermore,17 potential active metabolites were discovered by targeting the motilin,dopamine D2,and the serotonin type 4(5-HT4)receptors,and part bioactivities were verified using molecular dynamic simulation and a zebrafish constipation model.This study extends the application of mass spectrometry(MS)to rapidly profile TCM-derived metabolites in vivo,which will help pharmacologists rapidly discover potent metabolites from a complex matrix.
基金Supported by the Jiangsu Province’s Colleges and Universities(Integration of Chinese and Western Medicine)the National Natural Science Foundation of China[Methanism on Molecular and Hippocampus-Thalamus Neurocircuitry of Rapid Prokinetic and Antidepressant by Shugan Following Acute Stress(81973589)The Relationship Between Stress,Ghrelin Signaling,Liver-Soothing and Antidepressant Prokinetic Mechanisms(81573797)。
文摘OBJECTIVE:To eliminate ineffective or interfering compounds from the eight absorbed compounds(8ACs),identify the primary ACs that represent the multifunctional therapeutic effects of Zhiqiao(Fructus Aurantii Submaturus)and Houpo(Cortex Magnoliae Officinalis)(FM),along with elucidating their underlying mechanisms.METHODS:Key multifunctional ACs were screened through ex vivo-to-in vitro extrapolation(ex vivo dose=serum concentration)and validated in vivo,with efficacy assessed via contribution(dose=FM content).Functional magnetic resonance imaging analyzed brain regions's blood oxygen level-dependent(BOLD)changes,and the molecular mechanisms were analyzed by transcriptome of the dentate gyrus(DG).RESULTS:The results showed that representative 2ACs(Nobiletin+Magnolol)and 3ACs(Nobiletin+Magnolol+Meranzin hydrate)screened on ex vivo experiment by the criteria of contribution ranking contributed 80.72%-126.74%of the antidepressant and prokinetic effects of the FM(improvement of depressive-like behaviours,gastrointestinal disorder,monoamine neurotransmitters,ghrelin,endocrine hormones,pro-inflammatory factors,oxidative stress indicators).In addition,3ACs demonstrated superiority over 2ACs in improving depression and levels of multiple stress mediators.Zhiqiao(Fructus Aurantii Submaturus)(FRA)reduced acute stress-induced hyperactivation of the cingulate cortex,thalamus,hypothalamus,and entorhinal cortex and elevated BOLD signaling in the insular cortex,temporal association cortex.Furthermore,FRA upregulated pathways of neurotransmitter receptor activity and serotonergic synaptic function and downregulated inflammation-related pathways at the genetic level.CONCLUSION:2ACs and 3ACs closely reflected FM's multifunctional effect in antidepressant and prokinetic.FRA restores stress-impaired neural connectivity in functional brain regions and improves synaptic plasticity and neurogenesis at the genetic level.
基金supported by Xi'an Jiaotong University(No.01380011)
文摘A novel high performance liquid chromatographic method was developed for the determination of 4-O- methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation. Plasma samples were treated by one-fold volume of methanol and acetonitrile to remove the interference proteins. A reverse phase column of SHIM- PACK VP-ODS （150 mm × 4.6 mm, 5.0 μm） was used to separate 4-O-methylhonokiol in the plasma samples. The detection limit of 4-O-methylhonokiol was 0.2 μg/L and the linear range was 0. 012 - 1. 536 mg/L. The good extraction recoveries were obtained for the spiked samples （84.7%, 89.3% and 87.7% for low, middle and high concentrations of added standards, respectively）. The relative standard deviation of intra-day and inter-day precisions was in the range from 0.6% to 13.5%. The pharmacokinetic study of 4-O-methylhonokiol was made and the results from the plasmaconcentration curve of 4-0-methylhonokiol showed a two-apartment open model. This work developed a sensitive, stable and rapid HPLC method for the determination of 4-O-methylhonokiol and the developed method has been successfully applied to a pharmacokinetic study of 4-O-methylhonokiol.
