OBJECTIVE This study investigated transcriptional regulation of the main chemical con.stituents of Polygonum multiflorum Thunb.including Stilbene Glucoside(THSG) and anthraquinone constituents(Emodin,Rhein,Aloeemodin,...OBJECTIVE This study investigated transcriptional regulation of the main chemical con.stituents of Polygonum multiflorum Thunb.including Stilbene Glucoside(THSG) and anthraquinone constituents(Emodin,Rhein,Aloeemodin,Chrysophanol and Physcion) and six potential liver injury constituents(gallic acid,quercetin,luteolin,kaempferol,resveratrol) on mediated by PXR CYP3A4.Early establishment of pregnane X receptor mediated CYP3A4 drug induced rapid screening technique was used to determine the effects of these constituents.METHODS First,effect of constituents on the cell activity was detected by MTS cell viability assay.IC50 was calculated.Second,the expression vector and reporter vector were co-transfected into Hep G2 cells,10 μmol·L^(-1) Rifampicin as a positive control,10 μmol·L^(-1) Ketoconazole as a negative control.After treated with different concentrations of(the an.thraquinone constituents concentrations were 2.5,5 and 10 μmol·L^(-1);the concentrations of Gallic Acid,Quercetin,Luteolin,Kaempferol,Apigenin,Resveratrol concentrations were 5,10 and 20 μmol·L^(-1)) for24 h,the cells were tested for dual luciferase activity.RESULTS The results show that the inhibitory ef.fect of THSG,Chrysophanol,Emodin,Rhein and Aloeemodin on CYP3A4 was inhibited by co-transfec.tion of pcDNA3.1 and pGL4.17-CYP3A4.The expression of pcDNA3.14-PXR and pGL4.17-CYP3A4 was induced by the four constituents.Besides,Emodin has a directly inducing effect.Four anthraqui.none constituentscan induce the effect of CYP3A4 by PXR,but Emodin can directly induce CYP3A4.THSG can inhibit CYP3A4,but in the presence of PXR plasmid can induce CYP3A4.For the six poten.tial liver injury constituents,results show that the plasmid pcDNA3.1 was cotransfected with pGL4.17-CYP3A4 regulation of Gallic Acid and Resveratrol on CYP3A4 inhibitory effects of Quercetin,Luteolin,Kaempferol have an induce effect;after pcDNA3.14-PXR and pGL4.17-CYP3A4 cotransfected,Quercetin,Luteolin,Kaempferol,Apigenin,Resveratrol have induced effect,three constituents' induc.tion effect had significant difference.CONCLUSION 12 kinds of Polygonum multiflorum Thunb.constit.uents have inhibitory or activating effects on CYP3A4,after the participation of PXR,9 components have induced effects on CYP3A4,and the induction effect of 6 components has significant difference.The results suggested that we should pay attention to potential drug interactions when combined with Polygonum multiflorum Thunb.,and improve safety and efficacy.展开更多
基金supported by Beijing Natural Science Fund(7164291)Major National S&T Program(2015ZX09501004-003-003)Chinese Medicine Industry Projects(201507004)
文摘OBJECTIVE This study investigated transcriptional regulation of the main chemical con.stituents of Polygonum multiflorum Thunb.including Stilbene Glucoside(THSG) and anthraquinone constituents(Emodin,Rhein,Aloeemodin,Chrysophanol and Physcion) and six potential liver injury constituents(gallic acid,quercetin,luteolin,kaempferol,resveratrol) on mediated by PXR CYP3A4.Early establishment of pregnane X receptor mediated CYP3A4 drug induced rapid screening technique was used to determine the effects of these constituents.METHODS First,effect of constituents on the cell activity was detected by MTS cell viability assay.IC50 was calculated.Second,the expression vector and reporter vector were co-transfected into Hep G2 cells,10 μmol·L^(-1) Rifampicin as a positive control,10 μmol·L^(-1) Ketoconazole as a negative control.After treated with different concentrations of(the an.thraquinone constituents concentrations were 2.5,5 and 10 μmol·L^(-1);the concentrations of Gallic Acid,Quercetin,Luteolin,Kaempferol,Apigenin,Resveratrol concentrations were 5,10 and 20 μmol·L^(-1)) for24 h,the cells were tested for dual luciferase activity.RESULTS The results show that the inhibitory ef.fect of THSG,Chrysophanol,Emodin,Rhein and Aloeemodin on CYP3A4 was inhibited by co-transfec.tion of pcDNA3.1 and pGL4.17-CYP3A4.The expression of pcDNA3.14-PXR and pGL4.17-CYP3A4 was induced by the four constituents.Besides,Emodin has a directly inducing effect.Four anthraqui.none constituentscan induce the effect of CYP3A4 by PXR,but Emodin can directly induce CYP3A4.THSG can inhibit CYP3A4,but in the presence of PXR plasmid can induce CYP3A4.For the six poten.tial liver injury constituents,results show that the plasmid pcDNA3.1 was cotransfected with pGL4.17-CYP3A4 regulation of Gallic Acid and Resveratrol on CYP3A4 inhibitory effects of Quercetin,Luteolin,Kaempferol have an induce effect;after pcDNA3.14-PXR and pGL4.17-CYP3A4 cotransfected,Quercetin,Luteolin,Kaempferol,Apigenin,Resveratrol have induced effect,three constituents' induc.tion effect had significant difference.CONCLUSION 12 kinds of Polygonum multiflorum Thunb.constit.uents have inhibitory or activating effects on CYP3A4,after the participation of PXR,9 components have induced effects on CYP3A4,and the induction effect of 6 components has significant difference.The results suggested that we should pay attention to potential drug interactions when combined with Polygonum multiflorum Thunb.,and improve safety and efficacy.
基金supported by Hubei Provincial Central Government Guided Local Science and Technology Development Special Project“Traditional Chinese Herbal Medicine Properties and Quality Evaluation Platform”(2020ZYYD030)Hubei Province Health and Family Planning Scientific Research Project(WJ2023Q016).
