Fermentation offers a promising route to enhance the bioactivity of medicinal plants,yet traditional processes often rely on complex,undefined microbiota that limit reproducibility and yield.Here,using a microbiome-gu...Fermentation offers a promising route to enhance the bioactivity of medicinal plants,yet traditional processes often rely on complex,undefined microbiota that limit reproducibility and yield.Here,using a microbiome-guided approach,we develop a defined fermentation system for Platycodon grandiflorus(PG)aimed at the tar-geted enrichment of its key bioactive saponins,particularly platycodin D(PD).Metagenomic analysis of naturally fermented PG pickle revealed a Firmicutes-dominated community(>65%)with Weissella and Enterococcus as core genera.TargetingβD glucosidase-a key enzyme for platycoside activation-we isolated 141 microbial strains and identified Leuconostoc mesenteroides ZS W34-4 as the top performer in monobacterial fermentation.This defined starter initially increased total saponins content to 57.76±3.52 mg/g in static fermentation,a 41.3%gain over natural fermentation,and elevated PD by 26.25%.We further integrated ultrasound assistance to optimize the fermentation process parameters,which ultimately raised total saponins content to a maximum of 60.4±1.1mg/g-a 4.92%improvement over the static pure-culture fermentation and a 47.9%increase compared to natural fermentation.This progression demonstrates a stepwise enhancement from strain screening to process optimization.The strain maintained stable productivity over five passages(P>0.05),demonstrating robust process controllability.Our study elucidates the role of L.mesenteroides in platycoside transformation and establishes a scalable,microbiome-informed strategy for fermenting PG.This work provides a defined microbial starter and a tunable fermentation platform to support the standardized,high-value production of traditional fermented medicinal foods.展开更多
AIMS: To develop an HPLC-MS/MS method for the quantification of platycodin D(PD) in rat plasma, and to acquire the main pharmacokinetic parameters of PD after oral administration of pure PD or of Platycodi Radix extra...AIMS: To develop an HPLC-MS/MS method for the quantification of platycodin D(PD) in rat plasma, and to acquire the main pharmacokinetic parameters of PD after oral administration of pure PD or of Platycodi Radix extract(PRE) containing PD. METHOD: Plasma samples were pretreated with solid-phase extraction using an Oasis HLB SPE cartridge. Madecassoside was used as the internal standard(IS). Chromatographic separation was achieved on an ODS column(100 mm × 2.1 mm i.d., 3.5 μm) with a mobile phase consisting of acetonitrile/water(30 : 70, V/V) containing 0.1 mmol L 1ammonium acetate at a flow rate of 0.25 mL min 1. The detection was performed on a triple quadruple tandem mass spectrometer using an electrospray ionization(ESI) source with a chromatographic run time of 3.0 min. The detection was operated by multiple reaction monitoring(MRM) of the transitions of m/z 1 223.6→469.2 for PD and of m/z 973.6→469.2 for madecassoside(IS), respectively. RESULTS: The calibration curve was linear from 5 to 2 000 ng mL 1(r2>0.99) with a lower limit of quantification(LLOQ) of 5 ng mL 1. The intra- and inter-day precision(relative standard deviation, RSD) values were below 15% and the accuracy(relative error, RE) was from 15% to +15% at three quality control(QC) levels. Plasma concentrations of PD were determined for 24 h after i.v. administration of PD, and oral administration of PD and PRE, respectively. The absolute oral bioavailability of PD in rats was found to be(0.48 ± 0.19)% when administered PD, and to be(1.81 ± 0.89)% when administered PRE. CONCLUSION: The developed HPLC-MS/MS method was successfully applied to assess the pharmacokinetic parameters and oral bioavailability of PD in rats after administration of PD and Platycodi Radix extract.展开更多
Platycodin D(PD), a triterpenoid saponin isolated from Platycodonis Radix, is a famous Chinese herbal medicine that has been shown to have anti-proliferative effects in several cancer cell lines. The aim of this study...Platycodin D(PD), a triterpenoid saponin isolated from Platycodonis Radix, is a famous Chinese herbal medicine that has been shown to have anti-proliferative effects in several cancer cell lines. The aim of this study was to determine the changes in cellular proteins after the treatment of hepatocellular carcinoma HepG2 cells with PD using proteomics approaches. The cell viability was determined using the MTT assay. The proteome was analyzed by two-dimensional difference gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blot analysis was used to confirm the expression of changed proteins. Our results showed that PD inhibited the proliferation of HepG2 cells in concentration- and time-dependent manners. Sixteen proteins were identified to be up-regulated in PD-treated HepG2 cells, including ATP5 H, OXCT1, KRT9, CCDC40, ERP29, RCN1, ZNF175, HNRNPH1, HSP27, PA2G4, PHB, BANF1, TPM3, ECH1, LGALS1, and MYL6. Three proteins(i.e., RPS12, EMG1, and KRT1) decreased in HepG2 cells after treatment with PD. The changes in HSP27 and PHB were further confirmed by Western blotting. In conclusion, our results shed new lights on the mechanisms of action for the anti-cancer activity of PD.展开更多
Objective:To study the transmembrane transport of chemical components of Chinese herbs and to explore the function of platycodin D (PD) on biomembranes.Methods:Interaction between PD and the dipalmitoylphosphatidylcho...Objective:To study the transmembrane transport of chemical components of Chinese herbs and to explore the function of platycodin D (PD) on biomembranes.Methods:Interaction between PD and the dipalmitoylphosphatidylcholine (DPPC) bilayer was reproduced by molecular dynamics simulation with the Martini force field.A model validation and methodological study were first performed,and were based on simulation investigations of transmembrane transport for three herbal compounds with distinct hydrophilic properties.Results:PD increased the mobility of the DPPC bilayer since its aglycone strongly interacted with the hydrophobic layer,which broke the structure of the gate layer,and weakened the ordered performance of hydrophobic tails.Conclusion:The Martini force field was successfully applied to the study of the interaction between herbal compounds and a biological membrane.By combining the dynamics equilibrium morphology,the distribution of drugs inside and outside the biomembrane,and the interaction sites of drugs on the DPPC bilayer,factors influencing transmembrane transport of drugs were elucidated and the function of platycodin D in a biological membrane was reproduced.展开更多
Qingfei Paidu decoction(QFPDD)has been extensively used in clinical treatments during the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)epidemic.SARS-CoV-2 primarily invades host cells via its spike(S)pro...Qingfei Paidu decoction(QFPDD)has been extensively used in clinical treatments during the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)epidemic.SARS-CoV-2 primarily invades host cells via its spike(S)protein binding to the angiotensin-converting enzyme 2(ACE2)on the cell membrane,mediating viral-host membrane fusion.Blocking viral entry is a crucial step in preventing infection,with the interaction between the S receptor binding domain(S-RBD)and ACE2 being a key antiviral target.Given that SARS-CoV-2 predominantly affects the respiratory system and approxi-mately 25%of patients suffering from corona virus disease 2019(COVID-19)with gastrointestinal symptoms,we are com-mitted to identifying more active ingredients in QFPDD that target the respiratory and gastrointestinal tracts of COVID-19 patients.Among medicinal plants,ephedra and liquorice derived from QFPDD,along with two other Chinese herbs,Platycodon grandiflorum and Radix Rhei Et Rhizome(rhubarb),have garnered our interest.These herbs have historically been used in traditional Chinese medicine(TCM)for treating infectious diseases with respiratory and digestive symp-toms.Here,we established a library containing all components of the four individual herbs gathered from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and performed structure-based virtual screening to identify potential ACE2/S-RBD inhibitors.Subsequently,we selected 10 ingredients from the top 30 candidates and evaluated their activities using a pseudovirus neutralization assay.Delphinidin and deapio platyco-din D(DPD)showed significant antiviral potential with half-maximal inhibitory concentration(IC_(50))values of 45.35 μM and 1.38 μM,respectively.Furthermore,delphinidin also inhibited the 3-chymotrypsin-like protease(3CL^(pro)),indicating its dual-viral target inhibitory potential.Notably,DPD effectively suppressed HCoV-229E replication in BEL-7402 cells.This study not only provides a strategy for rapid identifying antiviral agents from TCM in anticipation of future pandemics but also offers theoretical and experimental evidence to support for the clinical use of QFPDD.展开更多
基金funded by the National Natural Science Foundation of China(Grant numbers U20A20400)Shenyang Agricultural University Talent Introduction(High Level)Research Initiation Fee Project(numbers 2023YJRC002)+4 种基金Liao Ning Revitalization Talents Program[XLYC2402005XLYC2213026]Liaoning Province Science and Technology Plan Project(2024JH2/101900005)Introduction of talents(high-level)research start-up fund project of Shenyang Agricultural University(2023YJRC002)Shenyang Science and technology innovation platform project[21-103-0-14,21-104-0-28].
文摘Fermentation offers a promising route to enhance the bioactivity of medicinal plants,yet traditional processes often rely on complex,undefined microbiota that limit reproducibility and yield.Here,using a microbiome-guided approach,we develop a defined fermentation system for Platycodon grandiflorus(PG)aimed at the tar-geted enrichment of its key bioactive saponins,particularly platycodin D(PD).Metagenomic analysis of naturally fermented PG pickle revealed a Firmicutes-dominated community(>65%)with Weissella and Enterococcus as core genera.TargetingβD glucosidase-a key enzyme for platycoside activation-we isolated 141 microbial strains and identified Leuconostoc mesenteroides ZS W34-4 as the top performer in monobacterial fermentation.This defined starter initially increased total saponins content to 57.76±3.52 mg/g in static fermentation,a 41.3%gain over natural fermentation,and elevated PD by 26.25%.We further integrated ultrasound assistance to optimize the fermentation process parameters,which ultimately raised total saponins content to a maximum of 60.4±1.1mg/g-a 4.92%improvement over the static pure-culture fermentation and a 47.9%increase compared to natural fermentation.This progression demonstrates a stepwise enhancement from strain screening to process optimization.The strain maintained stable productivity over five passages(P>0.05),demonstrating robust process controllability.Our study elucidates the role of L.mesenteroides in platycoside transformation and establishes a scalable,microbiome-informed strategy for fermenting PG.This work provides a defined microbial starter and a tunable fermentation platform to support the standardized,high-value production of traditional fermented medicinal foods.
基金supported by the National Natural Science Foundation of China(No.81073030)the National Key Technology R&D Program in the 11th Five Year Plan of China(No.2008BA151B00-2)
文摘AIMS: To develop an HPLC-MS/MS method for the quantification of platycodin D(PD) in rat plasma, and to acquire the main pharmacokinetic parameters of PD after oral administration of pure PD or of Platycodi Radix extract(PRE) containing PD. METHOD: Plasma samples were pretreated with solid-phase extraction using an Oasis HLB SPE cartridge. Madecassoside was used as the internal standard(IS). Chromatographic separation was achieved on an ODS column(100 mm × 2.1 mm i.d., 3.5 μm) with a mobile phase consisting of acetonitrile/water(30 : 70, V/V) containing 0.1 mmol L 1ammonium acetate at a flow rate of 0.25 mL min 1. The detection was performed on a triple quadruple tandem mass spectrometer using an electrospray ionization(ESI) source with a chromatographic run time of 3.0 min. The detection was operated by multiple reaction monitoring(MRM) of the transitions of m/z 1 223.6→469.2 for PD and of m/z 973.6→469.2 for madecassoside(IS), respectively. RESULTS: The calibration curve was linear from 5 to 2 000 ng mL 1(r2>0.99) with a lower limit of quantification(LLOQ) of 5 ng mL 1. The intra- and inter-day precision(relative standard deviation, RSD) values were below 15% and the accuracy(relative error, RE) was from 15% to +15% at three quality control(QC) levels. Plasma concentrations of PD were determined for 24 h after i.v. administration of PD, and oral administration of PD and PRE, respectively. The absolute oral bioavailability of PD in rats was found to be(0.48 ± 0.19)% when administered PD, and to be(1.81 ± 0.89)% when administered PRE. CONCLUSION: The developed HPLC-MS/MS method was successfully applied to assess the pharmacokinetic parameters and oral bioavailability of PD in rats after administration of PD and Platycodi Radix extract.
基金supported by the Science and Technology Development Fund,Macao S.A.R(FDCT)(070/2013/A)the Research Fund of University of Macao(SRG026-ICMS13-LJJ and MRG008-LJJ2014-ICMS)+1 种基金the Administration of Traditional Chinese Medicine of Zhejiang Province(No.2012 ZA028)Program of Xinmiao Talents in Zhejiang Province(No.2010 R410024)
文摘Platycodin D(PD), a triterpenoid saponin isolated from Platycodonis Radix, is a famous Chinese herbal medicine that has been shown to have anti-proliferative effects in several cancer cell lines. The aim of this study was to determine the changes in cellular proteins after the treatment of hepatocellular carcinoma HepG2 cells with PD using proteomics approaches. The cell viability was determined using the MTT assay. The proteome was analyzed by two-dimensional difference gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blot analysis was used to confirm the expression of changed proteins. Our results showed that PD inhibited the proliferation of HepG2 cells in concentration- and time-dependent manners. Sixteen proteins were identified to be up-regulated in PD-treated HepG2 cells, including ATP5 H, OXCT1, KRT9, CCDC40, ERP29, RCN1, ZNF175, HNRNPH1, HSP27, PA2G4, PHB, BANF1, TPM3, ECH1, LGALS1, and MYL6. Three proteins(i.e., RPS12, EMG1, and KRT1) decreased in HepG2 cells after treatment with PD. The changes in HSP27 and PHB were further confirmed by Western blotting. In conclusion, our results shed new lights on the mechanisms of action for the anti-cancer activity of PD.
基金The experiment was financially supported by the National Natural Science Foundation of China(81473364)Beijing Natural Science Foundation(7162122)Excellent Talents Training Subsidy Scheme of Beijing(2013D009999000003).
文摘Objective:To study the transmembrane transport of chemical components of Chinese herbs and to explore the function of platycodin D (PD) on biomembranes.Methods:Interaction between PD and the dipalmitoylphosphatidylcholine (DPPC) bilayer was reproduced by molecular dynamics simulation with the Martini force field.A model validation and methodological study were first performed,and were based on simulation investigations of transmembrane transport for three herbal compounds with distinct hydrophilic properties.Results:PD increased the mobility of the DPPC bilayer since its aglycone strongly interacted with the hydrophobic layer,which broke the structure of the gate layer,and weakened the ordered performance of hydrophobic tails.Conclusion:The Martini force field was successfully applied to the study of the interaction between herbal compounds and a biological membrane.By combining the dynamics equilibrium morphology,the distribution of drugs inside and outside the biomembrane,and the interaction sites of drugs on the DPPC bilayer,factors influencing transmembrane transport of drugs were elucidated and the function of platycodin D in a biological membrane was reproduced.
基金supported by the Shanghai Municipal Science and Technology Major Project(ZD2021CY001)the Shanghai Oriental Talent Program for Top-notch Project(No.BJWS2024068)+2 种基金the National Key Research and Development Program of China(2023YFC3503400,2022YFC3502000)the SIMM-SHUTCM Traditional Chinese Medicine Innovation Joint Research Program(E2G805H)the National Natural Science Foundation of China(82141203).
文摘Qingfei Paidu decoction(QFPDD)has been extensively used in clinical treatments during the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)epidemic.SARS-CoV-2 primarily invades host cells via its spike(S)protein binding to the angiotensin-converting enzyme 2(ACE2)on the cell membrane,mediating viral-host membrane fusion.Blocking viral entry is a crucial step in preventing infection,with the interaction between the S receptor binding domain(S-RBD)and ACE2 being a key antiviral target.Given that SARS-CoV-2 predominantly affects the respiratory system and approxi-mately 25%of patients suffering from corona virus disease 2019(COVID-19)with gastrointestinal symptoms,we are com-mitted to identifying more active ingredients in QFPDD that target the respiratory and gastrointestinal tracts of COVID-19 patients.Among medicinal plants,ephedra and liquorice derived from QFPDD,along with two other Chinese herbs,Platycodon grandiflorum and Radix Rhei Et Rhizome(rhubarb),have garnered our interest.These herbs have historically been used in traditional Chinese medicine(TCM)for treating infectious diseases with respiratory and digestive symp-toms.Here,we established a library containing all components of the four individual herbs gathered from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and performed structure-based virtual screening to identify potential ACE2/S-RBD inhibitors.Subsequently,we selected 10 ingredients from the top 30 candidates and evaluated their activities using a pseudovirus neutralization assay.Delphinidin and deapio platyco-din D(DPD)showed significant antiviral potential with half-maximal inhibitory concentration(IC_(50))values of 45.35 μM and 1.38 μM,respectively.Furthermore,delphinidin also inhibited the 3-chymotrypsin-like protease(3CL^(pro)),indicating its dual-viral target inhibitory potential.Notably,DPD effectively suppressed HCoV-229E replication in BEL-7402 cells.This study not only provides a strategy for rapid identifying antiviral agents from TCM in anticipation of future pandemics but also offers theoretical and experimental evidence to support for the clinical use of QFPDD.
