Objective:To develop an interference-free and rapid method to elucidate GuanxinⅡ(GXⅡ)'s representative vasodilator absorbed bioactive compounds(ABCs)among enormous phytochemicals.Methods:The contents of ferulic ...Objective:To develop an interference-free and rapid method to elucidate GuanxinⅡ(GXⅡ)'s representative vasodilator absorbed bioactive compounds(ABCs)among enormous phytochemicals.Methods:The contents of ferulic acid,tanshinol,and hydroxysafflor yellow A(FTA)in GXⅡ/rat serum after the oral administration of GXⅡ(30 g/kg)were detected using ultra-performance liquid chromatography-mass spectrometry.Totally 18 rats were randomly assigned to the control group(0.9%normal saline),GXⅡ(30 g/kg)and FTA(5,28 and 77 mg/kg)by random number table method.Diastolic coronary flow velocity-time integral(VTI),i.e.,coronary flow or coronary flow-mediated dilation(CFMD),and endothelium-intact vascular tension of isolated aortic rings were measured.After 12 h of exposure to blank medium or 0.5 mmol/L H_(2)O_(2),endothelial cells(ECs)were treated with post-dose GXⅡof supernatant from deproteinized serum(PGSDS,300μL PGSDS per 1 m L of culture medium)or FTA(237,1539,and 1510 mg/m L)for 10 min as control,H_(2)O_(2),PGSDS and FTA groups.Nitric oxide(NO),vascular endothelial growth factor(VEGF),endothelin-1(ET-1),superoxide dismutase(SOD),malondialdehyde(MDA)and phosphorylated phosphoinositide 3 kinase(p-PI3K),phosphorylated protein kinase B(p-AKT),phosphorylated endothelial nitric oxide synthase(p-e NOS)were analyzed.PGSDS was developed as a GXⅡproxy of ex vivo herbal crude extracts.Results:PGSDS effectively eliminates false responses caused by crude GXⅡpreparations.When doses equaled the contents in GXⅡ/its post-dose serum,FTA accounted for 98.17%of GXⅡ-added CFMD and 92.99%of PGSDS-reduced vascular tension.In ECs,FTA/PGSDS was found to have significant antioxidant(lower MDA and higher SOD,P<0.01)and endothelial function-protective(lower VEGF,ET-1,P<0.01)effects.The increases in aortic relaxation,endothelial NO levels and phosphorylated PI3K/Akt/e NOS protein induced by FTA/PGSDS were markedly abolished by NG-nitro-L-arginine methyl ester(L-NA,e NOS inhibitor)and wortmannin(PI3K/AKT inhibitor),respectively,indicating an endothelium-dependent vasodilation via the PI3K/AKT-e NOS pathway(P<0.01).Conclusion:This study provides a strategy for rapidly and precisely elucidating GXⅡ's representative in/ex vivo cardioprotective absorbed bioactive compounds(ABCs)-FTA,suggesting its potential in advancing precision ethnomedicine.展开更多
Objective:To study the pharmacological mechanism of Guanxin Ⅱ formula(冠心Ⅱ号方,GXⅡ)for treatment of coronary heart disease(CHD).Methods:A network pharmacology-based method was utilized.First candidate compounds,ta...Objective:To study the pharmacological mechanism of Guanxin Ⅱ formula(冠心Ⅱ号方,GXⅡ)for treatment of coronary heart disease(CHD).Methods:A network pharmacology-based method was utilized.First candidate compounds,targets of GXⅡ were collected using Pharm Mapper,BATMAN-TCM,Drug Bank and Swiss Target Prediction,and targets on CHD were mined from Gene Cards,Dis Genet,Drug Bank and GEO.Afterwards,the big hub compounds and targets were chosen in the candidate compounds-direct therapeutic targets on the CHD(C-T)network and the direct therapeutic targets on the CHD(T-D)network.Furthermore,the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis were performed to identify the enriched terms.Finally,a molecular docking simulation strategy was adopted to verify the binding capacity between the big hub compounds and big hub targets on CHD.Results:First,114 candidate compounds were selected with the following criteria:OB 30%,DL 0.18,and HL 4 h.Then,1,035 targets of GXⅡ were gathered,while 928 targets on CHD were collected.Afterwards,196 common targets of compound targets and therapeutic targets on CHD were defined as direct therapeutic targets acting on CHD.In addition,the contribution index(CI)in the C-T network was calculated,and 4 centrality properties,including degree,betweenness,closeness and coreness,in the T-D network,4 big hub compounds,and 6 big hub targets were eventually chosen.Furthermore,the GO and KEGG analysis indicated that GXⅡ acted on CHD by regulating the reactive oxygen species metabolism,steroid metabolism,lipid metabolism,inflammatory response,proliferation,differentiation and apoptosis.The docking results manifested excellent binding capacity between the 4 big hub compounds and 6 big hub targets on CHD.Conclusion:This network pharmacology-based exploration revealed that GXⅡ might prevent and inhibit the primary pathological processes of CHD.展开更多
基金Supported by the National Natural Science Foundation of China (No.81973589)。
文摘Objective:To develop an interference-free and rapid method to elucidate GuanxinⅡ(GXⅡ)'s representative vasodilator absorbed bioactive compounds(ABCs)among enormous phytochemicals.Methods:The contents of ferulic acid,tanshinol,and hydroxysafflor yellow A(FTA)in GXⅡ/rat serum after the oral administration of GXⅡ(30 g/kg)were detected using ultra-performance liquid chromatography-mass spectrometry.Totally 18 rats were randomly assigned to the control group(0.9%normal saline),GXⅡ(30 g/kg)and FTA(5,28 and 77 mg/kg)by random number table method.Diastolic coronary flow velocity-time integral(VTI),i.e.,coronary flow or coronary flow-mediated dilation(CFMD),and endothelium-intact vascular tension of isolated aortic rings were measured.After 12 h of exposure to blank medium or 0.5 mmol/L H_(2)O_(2),endothelial cells(ECs)were treated with post-dose GXⅡof supernatant from deproteinized serum(PGSDS,300μL PGSDS per 1 m L of culture medium)or FTA(237,1539,and 1510 mg/m L)for 10 min as control,H_(2)O_(2),PGSDS and FTA groups.Nitric oxide(NO),vascular endothelial growth factor(VEGF),endothelin-1(ET-1),superoxide dismutase(SOD),malondialdehyde(MDA)and phosphorylated phosphoinositide 3 kinase(p-PI3K),phosphorylated protein kinase B(p-AKT),phosphorylated endothelial nitric oxide synthase(p-e NOS)were analyzed.PGSDS was developed as a GXⅡproxy of ex vivo herbal crude extracts.Results:PGSDS effectively eliminates false responses caused by crude GXⅡpreparations.When doses equaled the contents in GXⅡ/its post-dose serum,FTA accounted for 98.17%of GXⅡ-added CFMD and 92.99%of PGSDS-reduced vascular tension.In ECs,FTA/PGSDS was found to have significant antioxidant(lower MDA and higher SOD,P<0.01)and endothelial function-protective(lower VEGF,ET-1,P<0.01)effects.The increases in aortic relaxation,endothelial NO levels and phosphorylated PI3K/Akt/e NOS protein induced by FTA/PGSDS were markedly abolished by NG-nitro-L-arginine methyl ester(L-NA,e NOS inhibitor)and wortmannin(PI3K/AKT inhibitor),respectively,indicating an endothelium-dependent vasodilation via the PI3K/AKT-e NOS pathway(P<0.01).Conclusion:This study provides a strategy for rapidly and precisely elucidating GXⅡ's representative in/ex vivo cardioprotective absorbed bioactive compounds(ABCs)-FTA,suggesting its potential in advancing precision ethnomedicine.
基金Supported by the National Natural Science Foundation of China(No.81573803)。
文摘Objective:To study the pharmacological mechanism of Guanxin Ⅱ formula(冠心Ⅱ号方,GXⅡ)for treatment of coronary heart disease(CHD).Methods:A network pharmacology-based method was utilized.First candidate compounds,targets of GXⅡ were collected using Pharm Mapper,BATMAN-TCM,Drug Bank and Swiss Target Prediction,and targets on CHD were mined from Gene Cards,Dis Genet,Drug Bank and GEO.Afterwards,the big hub compounds and targets were chosen in the candidate compounds-direct therapeutic targets on the CHD(C-T)network and the direct therapeutic targets on the CHD(T-D)network.Furthermore,the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis were performed to identify the enriched terms.Finally,a molecular docking simulation strategy was adopted to verify the binding capacity between the big hub compounds and big hub targets on CHD.Results:First,114 candidate compounds were selected with the following criteria:OB 30%,DL 0.18,and HL 4 h.Then,1,035 targets of GXⅡ were gathered,while 928 targets on CHD were collected.Afterwards,196 common targets of compound targets and therapeutic targets on CHD were defined as direct therapeutic targets acting on CHD.In addition,the contribution index(CI)in the C-T network was calculated,and 4 centrality properties,including degree,betweenness,closeness and coreness,in the T-D network,4 big hub compounds,and 6 big hub targets were eventually chosen.Furthermore,the GO and KEGG analysis indicated that GXⅡ acted on CHD by regulating the reactive oxygen species metabolism,steroid metabolism,lipid metabolism,inflammatory response,proliferation,differentiation and apoptosis.The docking results manifested excellent binding capacity between the 4 big hub compounds and 6 big hub targets on CHD.Conclusion:This network pharmacology-based exploration revealed that GXⅡ might prevent and inhibit the primary pathological processes of CHD.