OBJECTIVE:To investigate the potential pharmacological mechanism of Danlou tablet(丹蒌片,DLT)with a long-term clinical application in the treatment of myocardial ischemia/reperfusion(I/R)injury through network pharmac...OBJECTIVE:To investigate the potential pharmacological mechanism of Danlou tablet(丹蒌片,DLT)with a long-term clinical application in the treatment of myocardial ischemia/reperfusion(I/R)injury through network pharmacology,molecular docking and experimental verification.METHODS:The main chemical ingredients in DLT were retrieved from the Traditional Chinese Medicine(TCM)System Pharmacology Database,the TCM information database,the bioinformatics analysis tool for molecular mechanism of TCM,and HERB database.Disease targets of I/R were accessed from the databases of Online Mendelian Inheritance in Man,Gene Cards,Therapeutic Target Database,and Dis Ge NET database.The overlaying genes of DLT and I/R were obtained from the Venny online platform.The core targets and proteinprotein interaction network were constructed and analyzed via the Search Tool for the Retrieval of Interacting Genes Proteins database and Cytoscape software.Furthermore,Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed by the Metascape platform.Based on the results,the component-target-pathway network was constructed and drafted via the Cytoscape software and the platform of Bioinformatics.Furthermore,we performed molecular docking to predict the binding information between chemical molecules and target proteins.Finally,oxygenglucose deprivation/recovery(OGD/R)-induced H9c2 cardiomyocytes were used to validate the results of network pharmacology in vitro.RESULTS:A total of 189 active chemical components in DLT and 849 correlative targets of I/R were screened.Of note,133 overlaying genes found from the Venny online platform were concentrated into 28 core genes.Furthermore,the GO and KEGG pathway enrichment analysis presented that DLT might participate in 42 types of GO molecular functions,747 types of GO biological processes,19 types of GO cellular components,and 140 kinds of pathways to treat I/R.In the component-targetpathway network,the indirect relationship between herbs and their possible effective pathways was clarified.Based on the molecular docking,we speculated that Baicalein-prostaglandin G/H synthase 2(PTGS2)with-3.24 kcal/mol,Luteolin-heat shock protein 90 alpha family class A member 1(HSP90AA1)with-3.22 kcal/mol,Baicalein-HSP90AA1 with-3.13 kcal/mol,and QuercetinHSP90AA1 with-3.05 kcal/mol possessed the strongest binding force of less than-3 kcal/mol,sequentially.Experimental verification showed that Quercetin,Luteolin,and Baicalein could increase the relative cell viability of OGD/R-stimulated cardiomyocytes,probably by suppressing PTGS2,and activating HSP90AA1 and estrogen receptor 1 expression.CONCLUSIONS:We predicted the potential active compounds as the material basis of DLT that may provide a new approach to elucidate the novel pharmacological mechanism underlying the treatment of cardiac I/R damage.展开更多
[Objectives] To explore the efficacy of Danlou Tablet( DLT) in the treatment of coronary heart disease( CHD) with phlegm and blood stasis syndrome and its effects on serum inflammatory factors. [Methods]One hundred an...[Objectives] To explore the efficacy of Danlou Tablet( DLT) in the treatment of coronary heart disease( CHD) with phlegm and blood stasis syndrome and its effects on serum inflammatory factors. [Methods]One hundred and ninety-seven patients with CHD and phlegm and blood stasis syndrome in our hospital from January 2016 to January 2018 were selected and randomly divided into two groups: control group( n =98) treated with aspirin plus atorvastatin,and research group( n =99) treated with DLT and aspirin plus atorvastatin for one month. The clinical efficacy and incidence of adverse reactions were observed. Serum secretory phospholipase A2( s PLA2),lipoprotein-associated phospholipase A2( LP-PLA2),oxidized low-density lipoprotein( ox-LDL),monocyte chemoattractant protein-1( MCP-1) and World Health Organization Quality of Life( WHOQOL-100) scores were compared before and after one month of treatment. [Results] The total effective rate was93. 94% in the research group,which was higher than that in the control group( 79. 59%,P < 0. 05);the levels of serum s PLA2,LP-PLA2,ox-LDL and MCP-1 in the research group were lower than those in the control group after one month of treatment( P < 0. 05). There was no statistical significance of the difference in the total incidence of adverse reactions between the research group and the control group( P > 0. 05).After one month of treatment,WHOQOL-100 scores were higher in two groups,which were higher in the research group than that in the control group( P < 0. 05). [Conclusions]DLT can significantly reduce the level of serum inflammatory factors,improve the quality of life in patients with CHD and phlegm and blood stasis syndrome.展开更多
Background： It has been shown that administration of statins reduced the risk of peri-procedural myocardial damage. However, it remains unclear whether Chinese medicine Danlou Tablet （~）~）, similar to statins, may...Background： It has been shown that administration of statins reduced the risk of peri-procedural myocardial damage. However, it remains unclear whether Chinese medicine Danlou Tablet （~）~）, similar to statins, may protect patients undergoing percutaneous coronary intervention （PCI） from peri-procedural myocardial damage. Objective： To demonstrate the hypothesis whether treatment with Danlou Tablet would improve clinical outcome in patients undergoing selective PCI with non-ST elevation acute coronary syndrome （NSTE-ACS） in China. Methods： Approximately 220 patients with unstable angina or non-ST-segment elevation myocardial infarction undergoing PCI will be enrolled and randomized to Danlou Tablet treatment （4.5 g/day for 2 days before intervention, with a further 4.5 g/day for 90 days thereafter） or placebo. All patients will not receive Danlou Tablet before procedure. The primary end point is to evaluate the incidence of cardiac death, myocardial infarction or unplanned re-hospitalization and revascularization after 30 days in patients undergoing selective PCI treated with Danlou Tablet compared with placebo. Secondary endpoints include the incidence of peri-procedural myocardial injury, 3-month clinical outcomes, the quality of life and Chinese medicine syndromes assessment. Conclusion： This study protocol will provide important evidence of Danlou Tablet treatment on the peri-procedural myocardial injury in patients with NSTE-ACS undergoing selective PCI, which may support a strategy of routine Danlou Tablet therapy to improve the clinical outcomes.展开更多
Objective:To detect whether Danlou Tablet(DLT)regulates the hypoxia-induced factor(HIF)-1α-angiopoietin-like 4(Angptl4)mRNA signaling pathway and explore the role of DLT in treating chronic intermittent hypoxia(CIH)-...Objective:To detect whether Danlou Tablet(DLT)regulates the hypoxia-induced factor(HIF)-1α-angiopoietin-like 4(Angptl4)mRNA signaling pathway and explore the role of DLT in treating chronic intermittent hypoxia(CIH)-induced dyslipidemia and arteriosclerosis.Methods:The mature adipocytes were obtained from3 T3-L1 cell culturation and allocated into 8 groups including control groups(Groups 1 and 5,0.1 mL of cell culture grade water);DLT groups(Groups 2 and 6,0.1 mL of 1,000μg/mL DLT submicron powder solution);dimethyloxalylglycine(DMOG)groups(Groups 3 and 7,DMOG and 0.1 mL of cell culture grade water);DMOG plus DLT groups(Groups 4 and 8,DMOG and 0.1 mL of 1,000μg/mL DLT submicron powder solution).Groups1-4 used mature adipocytes and groups 5-8 used HIF-1α-siRNA lentivirus-transfected mature adipocytes.After 24-h treatment,real-time polymerase chain reaction and Western blot were employed to determine the mRNA and protein expression levels of HIF-1αand Angptl4.In animal experiments,the CIH model in ApoE^(-/-)mice was established.Sixteen mice were complete randomly divided into 4 groups including sham group,CIH model group[intermittent hypoxia and normal saline(2 mL/time)gavage once a day],Angptl4 Ab group[intermittent hypoxia and Angptl4 antibody(30 mg/kg)intraperitoneally injected every week],DLT group[intermittent hypoxia and DLT(250 mg/kg)once a day],4 mice in each group.After 4-week treatment,enzyme linked immunosorbent assay was used to detect the levels of serum total cholesterol(TC)and triglyceride(TG).Hematoxylin-eosin and CD68 staining were used to observe the morphological properties of arterial plaques.Results:Angptl4 expression was dependent on HIF-1α,with a reduction in mRNA expression and no response in protein level to DMOG or DLT treatment in relation to siHIF-1α-transfected cells.DLT inhibited HIF-1αand Angptl4 mRNA expression(P<0.05 or P<0.01)and reduced HIF-1αand Angptl4 protein expressions with DMOG in mature adipocytes(all P<0.01),as the effect on HIF-1αprotein also existed in the presence of siHIF-1α(P<0.01).ApoE^(-/-)mice treated with CIH had increased TG and TC levels(all P<0.01)and atherosclerotic plaque.Angptl4 antibody and DLT both reduced TG and TC levels(all P<0.01),as well as reducing atherosclerotic plaque areas,narrowing arterial wall thickness and alleviating atherosclerotic lesion symptoms to some extent.Conclusion:DLT had positive effects in improving dyslipidemia and arteriosclerosis by inhibiting Angptl4 protein level through HIF-1α-Angptl4 mRNA signaling pathway.展开更多
Objective： To observe the in vivo effect of Danlou Tablet（丹蒌片,DLT） on myocardial ischemia and reperfusion（I/R） injury.Methods： DLT effects were evaluated in mouse heart preparation using 30-min coronary occlu...Objective： To observe the in vivo effect of Danlou Tablet（丹蒌片,DLT） on myocardial ischemia and reperfusion（I/R） injury.Methods： DLT effects were evaluated in mouse heart preparation using 30-min coronary occlusion followed by 24-h reperfusion and compared among sham group（n=6）,I/R group（n=8）,IPC group（ischemia preconditioning,n=6） and DLT group（I/R with DLT pretreatment for 3 days,750 mg·kg^（-1）·day^（-1）,n=8）.The effects of DLT were characterized in infarction size（IS） compared with risk region（RR） and left ventricle using the Evans blue/triphenyltetrazolium chloride double dye staining method in vivo.Furthermore,the dose-dependent effect of DLT on I/R injury was evaluated by double staining method.Five different concentrations of DLT（0.625,1.25,2.5,5 and 10 g·kg^（-1）·day^（-1）） were chosen in this study,and dose-response curve of DLT was obtained on these data.Results： The ratio of IS to left ventricle was significantly smaller in the DLT and IPC groups than the I/R group（P〈0.05 or P〈0.01）,the ratio of IS to RR was also reduced in the DLT and IPC groups（P〈0.01）,while there were no differences in RR among the four groups（P〉0.05）.Experiments showed incidence of arrhythmias was reduced in the DLT group（P〈0.01）.Furthermore,DLT produced a dose-dependent inhibitory effect with a half maximal inhibitory concentration of 1.225 g·kg^（-1）·day^（-1）.Conclusions： Our research concluded that DLT was effective in reducing I/R injury in mice,and provided experimental supports for the clinical use of DLT.展开更多
文摘OBJECTIVE:To investigate the potential pharmacological mechanism of Danlou tablet(丹蒌片,DLT)with a long-term clinical application in the treatment of myocardial ischemia/reperfusion(I/R)injury through network pharmacology,molecular docking and experimental verification.METHODS:The main chemical ingredients in DLT were retrieved from the Traditional Chinese Medicine(TCM)System Pharmacology Database,the TCM information database,the bioinformatics analysis tool for molecular mechanism of TCM,and HERB database.Disease targets of I/R were accessed from the databases of Online Mendelian Inheritance in Man,Gene Cards,Therapeutic Target Database,and Dis Ge NET database.The overlaying genes of DLT and I/R were obtained from the Venny online platform.The core targets and proteinprotein interaction network were constructed and analyzed via the Search Tool for the Retrieval of Interacting Genes Proteins database and Cytoscape software.Furthermore,Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed by the Metascape platform.Based on the results,the component-target-pathway network was constructed and drafted via the Cytoscape software and the platform of Bioinformatics.Furthermore,we performed molecular docking to predict the binding information between chemical molecules and target proteins.Finally,oxygenglucose deprivation/recovery(OGD/R)-induced H9c2 cardiomyocytes were used to validate the results of network pharmacology in vitro.RESULTS:A total of 189 active chemical components in DLT and 849 correlative targets of I/R were screened.Of note,133 overlaying genes found from the Venny online platform were concentrated into 28 core genes.Furthermore,the GO and KEGG pathway enrichment analysis presented that DLT might participate in 42 types of GO molecular functions,747 types of GO biological processes,19 types of GO cellular components,and 140 kinds of pathways to treat I/R.In the component-targetpathway network,the indirect relationship between herbs and their possible effective pathways was clarified.Based on the molecular docking,we speculated that Baicalein-prostaglandin G/H synthase 2(PTGS2)with-3.24 kcal/mol,Luteolin-heat shock protein 90 alpha family class A member 1(HSP90AA1)with-3.22 kcal/mol,Baicalein-HSP90AA1 with-3.13 kcal/mol,and QuercetinHSP90AA1 with-3.05 kcal/mol possessed the strongest binding force of less than-3 kcal/mol,sequentially.Experimental verification showed that Quercetin,Luteolin,and Baicalein could increase the relative cell viability of OGD/R-stimulated cardiomyocytes,probably by suppressing PTGS2,and activating HSP90AA1 and estrogen receptor 1 expression.CONCLUSIONS:We predicted the potential active compounds as the material basis of DLT that may provide a new approach to elucidate the novel pharmacological mechanism underlying the treatment of cardiac I/R damage.
基金Supported by the Project of Shaanxi Provincial Science and Technology Department(2016TZC-S-14-3)
文摘[Objectives] To explore the efficacy of Danlou Tablet( DLT) in the treatment of coronary heart disease( CHD) with phlegm and blood stasis syndrome and its effects on serum inflammatory factors. [Methods]One hundred and ninety-seven patients with CHD and phlegm and blood stasis syndrome in our hospital from January 2016 to January 2018 were selected and randomly divided into two groups: control group( n =98) treated with aspirin plus atorvastatin,and research group( n =99) treated with DLT and aspirin plus atorvastatin for one month. The clinical efficacy and incidence of adverse reactions were observed. Serum secretory phospholipase A2( s PLA2),lipoprotein-associated phospholipase A2( LP-PLA2),oxidized low-density lipoprotein( ox-LDL),monocyte chemoattractant protein-1( MCP-1) and World Health Organization Quality of Life( WHOQOL-100) scores were compared before and after one month of treatment. [Results] The total effective rate was93. 94% in the research group,which was higher than that in the control group( 79. 59%,P < 0. 05);the levels of serum s PLA2,LP-PLA2,ox-LDL and MCP-1 in the research group were lower than those in the control group after one month of treatment( P < 0. 05). There was no statistical significance of the difference in the total incidence of adverse reactions between the research group and the control group( P > 0. 05).After one month of treatment,WHOQOL-100 scores were higher in two groups,which were higher in the research group than that in the control group( P < 0. 05). [Conclusions]DLT can significantly reduce the level of serum inflammatory factors,improve the quality of life in patients with CHD and phlegm and blood stasis syndrome.
基金Supported by the National Natural Science Foundation of China(No.81202782)a grant from the Department of Science and Technology and the Academy of Traditional Chinese Medicine of Guangdong Province,China(No.2012A032500013)
文摘Background： It has been shown that administration of statins reduced the risk of peri-procedural myocardial damage. However, it remains unclear whether Chinese medicine Danlou Tablet （~）~）, similar to statins, may protect patients undergoing percutaneous coronary intervention （PCI） from peri-procedural myocardial damage. Objective： To demonstrate the hypothesis whether treatment with Danlou Tablet would improve clinical outcome in patients undergoing selective PCI with non-ST elevation acute coronary syndrome （NSTE-ACS） in China. Methods： Approximately 220 patients with unstable angina or non-ST-segment elevation myocardial infarction undergoing PCI will be enrolled and randomized to Danlou Tablet treatment （4.5 g/day for 2 days before intervention, with a further 4.5 g/day for 90 days thereafter） or placebo. All patients will not receive Danlou Tablet before procedure. The primary end point is to evaluate the incidence of cardiac death, myocardial infarction or unplanned re-hospitalization and revascularization after 30 days in patients undergoing selective PCI treated with Danlou Tablet compared with placebo. Secondary endpoints include the incidence of peri-procedural myocardial injury, 3-month clinical outcomes, the quality of life and Chinese medicine syndromes assessment. Conclusion： This study protocol will provide important evidence of Danlou Tablet treatment on the peri-procedural myocardial injury in patients with NSTE-ACS undergoing selective PCI, which may support a strategy of routine Danlou Tablet therapy to improve the clinical outcomes.
基金Supported by National Natural Science Foundation of China(No.81473465)。
文摘Objective:To detect whether Danlou Tablet(DLT)regulates the hypoxia-induced factor(HIF)-1α-angiopoietin-like 4(Angptl4)mRNA signaling pathway and explore the role of DLT in treating chronic intermittent hypoxia(CIH)-induced dyslipidemia and arteriosclerosis.Methods:The mature adipocytes were obtained from3 T3-L1 cell culturation and allocated into 8 groups including control groups(Groups 1 and 5,0.1 mL of cell culture grade water);DLT groups(Groups 2 and 6,0.1 mL of 1,000μg/mL DLT submicron powder solution);dimethyloxalylglycine(DMOG)groups(Groups 3 and 7,DMOG and 0.1 mL of cell culture grade water);DMOG plus DLT groups(Groups 4 and 8,DMOG and 0.1 mL of 1,000μg/mL DLT submicron powder solution).Groups1-4 used mature adipocytes and groups 5-8 used HIF-1α-siRNA lentivirus-transfected mature adipocytes.After 24-h treatment,real-time polymerase chain reaction and Western blot were employed to determine the mRNA and protein expression levels of HIF-1αand Angptl4.In animal experiments,the CIH model in ApoE^(-/-)mice was established.Sixteen mice were complete randomly divided into 4 groups including sham group,CIH model group[intermittent hypoxia and normal saline(2 mL/time)gavage once a day],Angptl4 Ab group[intermittent hypoxia and Angptl4 antibody(30 mg/kg)intraperitoneally injected every week],DLT group[intermittent hypoxia and DLT(250 mg/kg)once a day],4 mice in each group.After 4-week treatment,enzyme linked immunosorbent assay was used to detect the levels of serum total cholesterol(TC)and triglyceride(TG).Hematoxylin-eosin and CD68 staining were used to observe the morphological properties of arterial plaques.Results:Angptl4 expression was dependent on HIF-1α,with a reduction in mRNA expression and no response in protein level to DMOG or DLT treatment in relation to siHIF-1α-transfected cells.DLT inhibited HIF-1αand Angptl4 mRNA expression(P<0.05 or P<0.01)and reduced HIF-1αand Angptl4 protein expressions with DMOG in mature adipocytes(all P<0.01),as the effect on HIF-1αprotein also existed in the presence of siHIF-1α(P<0.01).ApoE^(-/-)mice treated with CIH had increased TG and TC levels(all P<0.01)and atherosclerotic plaque.Angptl4 antibody and DLT both reduced TG and TC levels(all P<0.01),as well as reducing atherosclerotic plaque areas,narrowing arterial wall thickness and alleviating atherosclerotic lesion symptoms to some extent.Conclusion:DLT had positive effects in improving dyslipidemia and arteriosclerosis by inhibiting Angptl4 protein level through HIF-1α-Angptl4 mRNA signaling pathway.
基金Supported by Guangdong National Scientific Funding(No.2014A030313402)Guangdong Medical Research Foundation(No.A2014271)National Natural Science Foundation of China(No.81473471 and No.81573708)
文摘Objective： To observe the in vivo effect of Danlou Tablet（丹蒌片,DLT） on myocardial ischemia and reperfusion（I/R） injury.Methods： DLT effects were evaluated in mouse heart preparation using 30-min coronary occlusion followed by 24-h reperfusion and compared among sham group（n=6）,I/R group（n=8）,IPC group（ischemia preconditioning,n=6） and DLT group（I/R with DLT pretreatment for 3 days,750 mg·kg^（-1）·day^（-1）,n=8）.The effects of DLT were characterized in infarction size（IS） compared with risk region（RR） and left ventricle using the Evans blue/triphenyltetrazolium chloride double dye staining method in vivo.Furthermore,the dose-dependent effect of DLT on I/R injury was evaluated by double staining method.Five different concentrations of DLT（0.625,1.25,2.5,5 and 10 g·kg^（-1）·day^（-1）） were chosen in this study,and dose-response curve of DLT was obtained on these data.Results： The ratio of IS to left ventricle was significantly smaller in the DLT and IPC groups than the I/R group（P〈0.05 or P〈0.01）,the ratio of IS to RR was also reduced in the DLT and IPC groups（P〈0.01）,while there were no differences in RR among the four groups（P〉0.05）.Experiments showed incidence of arrhythmias was reduced in the DLT group（P〈0.01）.Furthermore,DLT produced a dose-dependent inhibitory effect with a half maximal inhibitory concentration of 1.225 g·kg^（-1）·day^（-1）.Conclusions： Our research concluded that DLT was effective in reducing I/R injury in mice,and provided experimental supports for the clinical use of DLT.
