Background:Valtrate(Val)was extracted from the Valeriana jatamansi Jones plant,had good antitumor activity.However,its precise molecular mechanism in cancer cells was still unclear.This study investigated the effect o...Background:Valtrate(Val)was extracted from the Valeriana jatamansi Jones plant,had good antitumor activity.However,its precise molecular mechanism in cancer cells was still unclear.This study investigated the effect of Val on gastric cancer(GC)cells and its potential molecular mechanism.Methods:Cell viability was examined by CCK-8 assay.Cell cycle,apoptosis,and Reactive oxygen species(ROS)level were analyzed by flow cytometry.The migration effect of Val on AGS cells was analyzed by transwell and wound-healing assay.The expression levels of proteins were analyzed by western blot.Results:The cell viability assay results demonstrated that Val significantly decreased GC cell viability.Apoptosis assay results revealed that Val induced mitochondria-dependent apoptosis through the Bad/Bcl-2/cyto-c/cle-casp-3/cle-PARP pathways.Further exploration found that Val induced apoptosis through increasing the expression of phosphorylated p38 mitogen-activated protein kinase(p-p38),phosphorylated c-Jun N-terminal kinase(p-JNK),and Inhibitor kappa B alpha(IκB-α)proteins and decreasing the expression of phosphorylated extracellular signal-regulated kinase(p-ERK),phosphorylated signal transducer and activator of transcription 3(p-STAT3),and nuclear factor kappa-B(NF-κB)proteins;these expression levels of proteins were reversed by mitogen-activated protein kinase(MAPK)inhibitor.Furthermore,Val induced G2/M phase arrest in AGS cells through downregulating the expression of phosphorylated protein kinase B(p-AKT).Moreover,Val induced inhibition of AGS cell migration through downregulating the expression of p-GSK-3βandβ-catenin.In addition,Val promoted the ROS accumulation of AGS cells.Further investigation found that Val-induced apoptosis,arrested the cell cycle,and inhibited cell migration,and that its signaling pathways related to protein expressions were reversed by the ROS scavenger,N-acetyl-L-cysteine.Conclusion:Val induced apoptosis,arrested the cell cycle,and inhibited migration by ROS-mediated MAPK/STAT3/NF-κB,AKT/Cyclin B/CDK1/2,and GSK-3β/β-catenin signaling pathways in AGS cells.展开更多
Valtrate is the main drug quality control for the qualitative and quantitative analysis of Valerian medicines in the Chinese Pharmacopoeia 2010. However, valtrate is unstable under some conditions. We, for the first t...Valtrate is the main drug quality control for the qualitative and quantitative analysis of Valerian medicines in the Chinese Pharmacopoeia 2010. However, valtrate is unstable under some conditions. We, for the first time, systemically evaluated the stability of two bath reference standards （RS） by high performance liquid chromatography coupled with a triple quadrupole mass spectrometer （HPLC-MS/MS）. The forced degradations of valtrate were performed to evaluate its optimal storage, transportation and experiment conditions according to ICH guideline. The developed HPLC method was validated to determine the degradation products. Valtrate RS was sensitive to alkaline and thermal conditions, but it was relatively stable under acidic, oxidation and photolysis conditions. A total of nine degradation components were identified under alkaline hydrolysis （N1-N4） and thermal degradation （B1-B5）. The information obtained in this work would be valuable to minimize the decomposition of valtrate during the processes of preparation, storage, distribution and utilization. It was highly suggested to store valtrate with a single dose packing in brown closed ampoule at -20℃. Under the above-mentioned storage condition, valtrate could be stable for up to 3 years.展开更多
基金funded by Central Government Supports Local College Reform and Development Fund Talent Training Project(No.2020GSP16)Heilongjiang Province Key Research and Development Plan Guidance Project(No.GZ20220039)+2 种基金Heilongjiang Touyan Innovation Team Program(No.2019HTY078)Program of Heilongjiang Bayi Agricultural University(No.XDB201818)Postgraduate Innovative Research Project of Heilongjiang Bayi Agricultural University(No.YJSCX2022-Y55).
文摘Background:Valtrate(Val)was extracted from the Valeriana jatamansi Jones plant,had good antitumor activity.However,its precise molecular mechanism in cancer cells was still unclear.This study investigated the effect of Val on gastric cancer(GC)cells and its potential molecular mechanism.Methods:Cell viability was examined by CCK-8 assay.Cell cycle,apoptosis,and Reactive oxygen species(ROS)level were analyzed by flow cytometry.The migration effect of Val on AGS cells was analyzed by transwell and wound-healing assay.The expression levels of proteins were analyzed by western blot.Results:The cell viability assay results demonstrated that Val significantly decreased GC cell viability.Apoptosis assay results revealed that Val induced mitochondria-dependent apoptosis through the Bad/Bcl-2/cyto-c/cle-casp-3/cle-PARP pathways.Further exploration found that Val induced apoptosis through increasing the expression of phosphorylated p38 mitogen-activated protein kinase(p-p38),phosphorylated c-Jun N-terminal kinase(p-JNK),and Inhibitor kappa B alpha(IκB-α)proteins and decreasing the expression of phosphorylated extracellular signal-regulated kinase(p-ERK),phosphorylated signal transducer and activator of transcription 3(p-STAT3),and nuclear factor kappa-B(NF-κB)proteins;these expression levels of proteins were reversed by mitogen-activated protein kinase(MAPK)inhibitor.Furthermore,Val induced G2/M phase arrest in AGS cells through downregulating the expression of phosphorylated protein kinase B(p-AKT).Moreover,Val induced inhibition of AGS cell migration through downregulating the expression of p-GSK-3βandβ-catenin.In addition,Val promoted the ROS accumulation of AGS cells.Further investigation found that Val-induced apoptosis,arrested the cell cycle,and inhibited cell migration,and that its signaling pathways related to protein expressions were reversed by the ROS scavenger,N-acetyl-L-cysteine.Conclusion:Val induced apoptosis,arrested the cell cycle,and inhibited migration by ROS-mediated MAPK/STAT3/NF-κB,AKT/Cyclin B/CDK1/2,and GSK-3β/β-catenin signaling pathways in AGS cells.
基金Special Funds of the National Natural Science Foundation of China on"Major New Drugs Innovation and Development"(Grant No.2014ZX09304307-002)
文摘Valtrate is the main drug quality control for the qualitative and quantitative analysis of Valerian medicines in the Chinese Pharmacopoeia 2010. However, valtrate is unstable under some conditions. We, for the first time, systemically evaluated the stability of two bath reference standards （RS） by high performance liquid chromatography coupled with a triple quadrupole mass spectrometer （HPLC-MS/MS）. The forced degradations of valtrate were performed to evaluate its optimal storage, transportation and experiment conditions according to ICH guideline. The developed HPLC method was validated to determine the degradation products. Valtrate RS was sensitive to alkaline and thermal conditions, but it was relatively stable under acidic, oxidation and photolysis conditions. A total of nine degradation components were identified under alkaline hydrolysis （N1-N4） and thermal degradation （B1-B5）. The information obtained in this work would be valuable to minimize the decomposition of valtrate during the processes of preparation, storage, distribution and utilization. It was highly suggested to store valtrate with a single dose packing in brown closed ampoule at -20℃. Under the above-mentioned storage condition, valtrate could be stable for up to 3 years.
