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
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.
