Objective To investigate whether Lingbao Huxin Pill(LBHX)protects against acute myocardial infarction(AMI)at the infarct border zone(IBZ)of myocardial tissue by regulating apoptosis and inflammation through the sirtui...Objective To investigate whether Lingbao Huxin Pill(LBHX)protects against acute myocardial infarction(AMI)at the infarct border zone(IBZ)of myocardial tissue by regulating apoptosis and inflammation through the sirtuin 1(SIRT1)-mediated forkhead box protein O1(FOXO1)and nuclear factor-κB(NF-κB)signaling pathways.Methods Six-week-old Wistar rats with normal diet were randomized into the sham,the model,Betaloc(0.9 mg/kg daily),LBHX-L(0.45 mg/kg daily),LBHX-M(0.9 mg/kg daily),LBHX-H(1.8 mg/kg daily),and LBHX+EX527(0.9 mg/kg daily)groups according to the method of random number table,13 in each group.In this study,left anterior descending coronary artery(LADCA)ligation was performed to induce an AMI model in rats.The myocardial infarction area was examined using a 2,3,5-triphenyltetrazolium chloride solution staining assay.A TdT-mediated dUTP nick-end labeling(TUNEL)assay was conducted to assess cardiomyocyte apoptosis in the IBZ.The histopathology of myocardial tissue at the IBZ was assessed with Heidenhain,Masson and hematoxylineosin(HE)staining assays.The expression levels of tumor necrosis factorα(TNF-α),interleukin(IL)-6,IL-1β,and intercellular adhesion molecule-1 were measured using enzyme-linked immunosorbent assays(ELISAs).The mRNA expressions of SIRT1 and FOXO1 were detected by real-time qPCR(RT-qPCR).The protein expressions of SIRT1,FOXO1,SOD2,BAX and NF-κB p65 were detected by Western blot analysis.Results The ligation of the LADCA successfully induced an AMI model.The LBHX pretreatment reduced the infarct size in the AMI rats(P<0.01).The TUNEL assay revealed that LBHX inhibited cardiomyocyte apoptosis at the IBZ.Further,the histological examination showed that the LBHX pretreatment decreased the ischemic area of myocardial tissue(P<0.05),myocardial interstitial collagen deposition(P<0.05)and inflammation at the IBZ.The ELISA results indicated that LBHX decreased the serum levels of inflammatory cytokines in the AMI rats(P<0.05 or P<0.01).Furthermore,Western blot analysis revealed that the LBHX pretreatment upregulated the protein levels of SIRT1,FOXO1 and SOD2(P<0.05)and downregulated NF-κB p65 and BAX expressions(P<0.05).The RT-qPCR results showed that LBHX increased the SIRT1 mRNA and FOXO1 mRNA levels(P<0.05).These protective effects,including inhibiting apoptosis and alleviating inflammation in the IBZ,were partially abolished by EX527,an inhibitor of SIRT1.Conclusion LBHX could protect against AMI by suppressing apoptosis and inflammation in AMI rats and the SIRT1-mediated FOXO1 and NF-κB signaling pathways were involved in the cardioprotection effect of LBHX.展开更多
OBJECTIVE:To investigate the mechanism of action of the Lingbao Huxin Dan(灵宝护心丹)in treating bradycardia arrhythmia with coronary heart disease(BACHD)by network pharmacology.METHODS:The active ingredients of the L...OBJECTIVE:To investigate the mechanism of action of the Lingbao Huxin Dan(灵宝护心丹)in treating bradycardia arrhythmia with coronary heart disease(BACHD)by network pharmacology.METHODS:The active ingredients of the Lingbao Huxin Dan were screened on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Bioinformatics tools designed for the analysis of molecular mechanisms of Chinese medicine platform;target prediction was conducted with the SwissTargetPrediction database,and Cytoscape 3.8 was used to construct a drug ingredient-target network.The Genecards,Online Mendelian Inheritance in Man,and DrugB ank databases were searched for disease targets.Venn plots were used to display the common targets of BA-CHD and active ingredients.The STRING platform was used to construct a protein-protein interaction network.The Metascape data platform was used for Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis to construct a signaling pathway network of the active ingredients of the Lingbao Huxin Dan.RESULTS:There were 121 active ingredients,899 related targets,39 targets important in BA-CHD and 14 targets which intersected between the active ingredients and BA-CHD.There were 27 core therapeutic ingredients,153 biological processes,18 cell ingredients and 20 molecular functions obtained by GO enrichment analysis.The KEGG pathway analysis yielded 19 signaling pathways.CONCLUSION:RBA-CHD may treat BA-CHD by regulating adrenergic receptor beta-1,alpha 1-αadrenergic receptor,calcium voltage-gated channel subunit alpha1 C,alpha-1β-adrenergic receptor,nitric oxide synthase 2,beta-2 adrenergic receptor,voltagedependent calcium channel subunit alpha-2/delta-1,angiotensin-converting enzyme,Raf-1 proto-oncogene serine/threonine-protein kinase,and other targets,potentially by affecting adrenergic receptor binding and calcium channel opening,to regulate the activity of cardiomyocytes.展开更多
基金Supported by the National Natural Science Foundation of China(No.81774141 and No.82074418)。
文摘Objective To investigate whether Lingbao Huxin Pill(LBHX)protects against acute myocardial infarction(AMI)at the infarct border zone(IBZ)of myocardial tissue by regulating apoptosis and inflammation through the sirtuin 1(SIRT1)-mediated forkhead box protein O1(FOXO1)and nuclear factor-κB(NF-κB)signaling pathways.Methods Six-week-old Wistar rats with normal diet were randomized into the sham,the model,Betaloc(0.9 mg/kg daily),LBHX-L(0.45 mg/kg daily),LBHX-M(0.9 mg/kg daily),LBHX-H(1.8 mg/kg daily),and LBHX+EX527(0.9 mg/kg daily)groups according to the method of random number table,13 in each group.In this study,left anterior descending coronary artery(LADCA)ligation was performed to induce an AMI model in rats.The myocardial infarction area was examined using a 2,3,5-triphenyltetrazolium chloride solution staining assay.A TdT-mediated dUTP nick-end labeling(TUNEL)assay was conducted to assess cardiomyocyte apoptosis in the IBZ.The histopathology of myocardial tissue at the IBZ was assessed with Heidenhain,Masson and hematoxylineosin(HE)staining assays.The expression levels of tumor necrosis factorα(TNF-α),interleukin(IL)-6,IL-1β,and intercellular adhesion molecule-1 were measured using enzyme-linked immunosorbent assays(ELISAs).The mRNA expressions of SIRT1 and FOXO1 were detected by real-time qPCR(RT-qPCR).The protein expressions of SIRT1,FOXO1,SOD2,BAX and NF-κB p65 were detected by Western blot analysis.Results The ligation of the LADCA successfully induced an AMI model.The LBHX pretreatment reduced the infarct size in the AMI rats(P<0.01).The TUNEL assay revealed that LBHX inhibited cardiomyocyte apoptosis at the IBZ.Further,the histological examination showed that the LBHX pretreatment decreased the ischemic area of myocardial tissue(P<0.05),myocardial interstitial collagen deposition(P<0.05)and inflammation at the IBZ.The ELISA results indicated that LBHX decreased the serum levels of inflammatory cytokines in the AMI rats(P<0.05 or P<0.01).Furthermore,Western blot analysis revealed that the LBHX pretreatment upregulated the protein levels of SIRT1,FOXO1 and SOD2(P<0.05)and downregulated NF-κB p65 and BAX expressions(P<0.05).The RT-qPCR results showed that LBHX increased the SIRT1 mRNA and FOXO1 mRNA levels(P<0.05).These protective effects,including inhibiting apoptosis and alleviating inflammation in the IBZ,were partially abolished by EX527,an inhibitor of SIRT1.Conclusion LBHX could protect against AMI by suppressing apoptosis and inflammation in AMI rats and the SIRT1-mediated FOXO1 and NF-κB signaling pathways were involved in the cardioprotection effect of LBHX.
基金Training Project of National Administration of Traditional Chinese Medicine“Natural Training Project of Backbone of Traditional Chinese Medicine”(SATCM,No.2019-128)the Fundamental Research Funds for the Central Public Welfare Research Institutes:the Application of Panoramic Tissue Scanning and Quantitative Analysis Systems in the Research of Traditional Chinese Medicine for Disease Prevention and Treatment”(No.JBGS2021005)。
文摘OBJECTIVE:To investigate the mechanism of action of the Lingbao Huxin Dan(灵宝护心丹)in treating bradycardia arrhythmia with coronary heart disease(BACHD)by network pharmacology.METHODS:The active ingredients of the Lingbao Huxin Dan were screened on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Bioinformatics tools designed for the analysis of molecular mechanisms of Chinese medicine platform;target prediction was conducted with the SwissTargetPrediction database,and Cytoscape 3.8 was used to construct a drug ingredient-target network.The Genecards,Online Mendelian Inheritance in Man,and DrugB ank databases were searched for disease targets.Venn plots were used to display the common targets of BA-CHD and active ingredients.The STRING platform was used to construct a protein-protein interaction network.The Metascape data platform was used for Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis to construct a signaling pathway network of the active ingredients of the Lingbao Huxin Dan.RESULTS:There were 121 active ingredients,899 related targets,39 targets important in BA-CHD and 14 targets which intersected between the active ingredients and BA-CHD.There were 27 core therapeutic ingredients,153 biological processes,18 cell ingredients and 20 molecular functions obtained by GO enrichment analysis.The KEGG pathway analysis yielded 19 signaling pathways.CONCLUSION:RBA-CHD may treat BA-CHD by regulating adrenergic receptor beta-1,alpha 1-αadrenergic receptor,calcium voltage-gated channel subunit alpha1 C,alpha-1β-adrenergic receptor,nitric oxide synthase 2,beta-2 adrenergic receptor,voltagedependent calcium channel subunit alpha-2/delta-1,angiotensin-converting enzyme,Raf-1 proto-oncogene serine/threonine-protein kinase,and other targets,potentially by affecting adrenergic receptor binding and calcium channel opening,to regulate the activity of cardiomyocytes.
