AIM:To investigate the hepatoprotective effects and mechanisms of an extract of Salvia miltiorrhiza and Carthamus tinctorius in vivo.METHODS:C57BL/6J mice were randomly assigned to five groups and intraperitoneally ad...AIM:To investigate the hepatoprotective effects and mechanisms of an extract of Salvia miltiorrhiza and Carthamus tinctorius in vivo.METHODS:C57BL/6J mice were randomly assigned to five groups and intraperitoneally administered 0.9% saline,Salvia miltiorrhiza and Carthamus tinctorius extract [Danhong injection(DHI),0.75 and 3 g/kg mixed extract] or reduced glutathione for injection(RGI,300 mg/kg) for 30 min before exposure to lipopolysaccharide(LPS,16 mg/kg). After intraperitoneal LPS stimulation for 90 min or 6 h,the mice were sacrificed by ether anaesthesia,and serum and liver samples were collected. Histological analysis(H&E) and terminal deoxynucleotidyl transferase-mediated d UTP nick end-labelling(TUNEL) staining were performed. Alanine transferase(ALT),aspartate transaminase(AST),total bilirubin(TBil),glutathione-S-transferase(GST),malondialdehyde(MDA),tumour necrosis factor(TNF)-α,interleukin(IL)-6,and caspase-3 levels were measured. Bax,Bcl-2,P-IκBα,IκBα,P-NF-κB p65,and NF-κB p65 protein levels were determined by Western blot. TNF-α,IL-6,caspase-3,Bax and Bcl-2 m RNA expression was measured by real-time reverse transcription-polymerase chain reaction(RT-PCR).RESULTS:Hematoxylin-eosin staining and TUNEL results suggested that DHI(3 g/kg) treatment alleviated inflammatory and apoptotic(P < 0.01) injury in the liver of mice. DHI treatment dose-dependently blunted the abnormal changes in biochemical parameters such as ALT(72.53 ± 2.83 for 3 g/kg,P < 0.01),AST(76.97 ± 5.00 for 3 g/kg,P < 0.01),TBil(1.17 ± 0.10 for 3 g/kg,P < 0.01),MDA(0.81 ± 0.36 for 3 g/kg,P < 0.01),and GST(358.86 ± 12.09 for 3 g/kg,P < 0.01). Moreover,DHI(3 g/kg) remarkably decreased LPS-induced protein expression of TNF-α(340.55 ± 10.18 for 3 g/kg,P < 0.01),IL-6(261.34 ± 10.18 for 3 g/kg,P < 0.01),and enzyme activity of caspase-3(0.93 ± 0.029 for 3 g/kg,P < 0.01). The LPS-induced m RNA expression of TNF-α,IL-6 and caspase-3 was also decreased by DHI. Western blot analysis revealed that DHI antagonised LPS-stimulated decrease of Bcl-2 and increase of Bax protein expression. Furthermore,DHI inhibited LPS-induced IκBα and NF-κB p65 phosphorylation.CONCLUSION:DHI may be a multi-function protectant against acute hepatic injury in mice through its antiinflammatory,anti-oxidative and anti-apoptotic activities.展开更多
Based on network pharmacology and molecular docking technology,the similarities and differences of anti-myocardial ischemia mechanism between Salvia miltiorrhiza Bunge(SM)and Carthamus tinctorius L.(CT)were studied.Fi...Based on network pharmacology and molecular docking technology,the similarities and differences of anti-myocardial ischemia mechanism between Salvia miltiorrhiza Bunge(SM)and Carthamus tinctorius L.(CT)were studied.Firstly,based on the traditional Chinese medicine systems pharmacology(TCMSP),the related compounds of SM and CT were obtained,and the potential targets of these compounds were collected by the target fishing method.Genecards database was used to obtain targets related to myocardial ischemia.The cross targets of CT,SM,and myocardial ischemia were then selected,and the protein-protein interaction(PPI)network was constructed based on the STRING database.The cross targets were imported into the Metascape database for Gene Oncology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.Cytoscape software was used to build the topological network diagram of the drug-compound-target path.Finally,the binding ability of the active ingredient and the key target was verified by molecular docking.65 active ingredients and 38 potential targets were screened from SM,and 22 active ingredients and 58 potential targets were excavated from CT.Important targets common to SM and CT were TNF,IL6,VEGFA,AKT1,etc.The common enrichment pathways involved are fluid shear stress and atherosclerosis,IL-17 signaling pathway,pathways in cancer,and toxoplasmosis.The findings suggested that the two traditional Chinese medicines exerted the effect of myocardial ischemia through the characteristics of multiple targets,multiple pathways,and multiple compounds.展开更多
目的基于网络药理学筛选丹参-红花药对配伍抗缺血性脑卒中的成分靶点、作用靶点,并对大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)大鼠模型进行相关作用靶点的实验验证。方法①通过系统药理学平台(Traditional Chinese Medic...目的基于网络药理学筛选丹参-红花药对配伍抗缺血性脑卒中的成分靶点、作用靶点,并对大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)大鼠模型进行相关作用靶点的实验验证。方法①通过系统药理学平台(Traditional Chinese Medicine Systems Pharmacology,TCMSP)、基因名片(GeneCards)数据库筛选丹参-红花的有效成分、成分靶点及缺血性脑卒中的相关靶点;运用STRING数据库构建蛋白互作网络图(protein protein interaction network,PPI),并利用生物信息分析软件(Cytoscape3.8.0)构建成分靶点互作网络;利用David在线分析工具对目标基因进行基因本体论(gene ontology,GO)富集分析和基因的京都百科全书和基因组(kyoto encyclopedia of genes and genomes,KEGG)通路分析。②采用改良MCAO法制备缺血性脑卒中大鼠模型,采用免疫组化法、实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠脑组织中NLR家族Pyrin域蛋白3(NLRP3)炎症小体和核转录因子-B(NF-kB)p65(NFkBp65)蛋白的阳性表达,探讨丹红配伍抗脑缺血损伤炎症反应的作用机制。结果①丹参-红花药对中筛选出87个有效成分,253个靶点,缺血性脑卒中疾病靶点1448个,药物与疾病相关靶点161个,通过GO分析得出730个生物学过程(biological process,BP),81个细胞组分(cellular component,CC),128个分子功能(molecular function,MF),通过KEGG分析得出127条信号通路。②免疫组化法和Real-time PCR测定结果显示,与假手术组比较,模型组大鼠脑组织中NLRP3炎症小体和NFkBp65蛋白表达明显增加(P<0.01),与模型组比较,丹红配伍组和尼莫地平组NLRP3炎症小体和NFkBp65蛋白表达明显降低(P<0.01)。结论丹红有效组分配伍能够通过NF-κB信号通路下调NLRP3炎症小体的释放,阻断炎症损伤环节,最终达到抗炎损伤的作用。展开更多
基金Supported by National Natural Science Foundation of China,No.81173469 and No.81273891the Key New Drug Creation and Manufacturing Program,No.2012ZX09304007
文摘AIM:To investigate the hepatoprotective effects and mechanisms of an extract of Salvia miltiorrhiza and Carthamus tinctorius in vivo.METHODS:C57BL/6J mice were randomly assigned to five groups and intraperitoneally administered 0.9% saline,Salvia miltiorrhiza and Carthamus tinctorius extract [Danhong injection(DHI),0.75 and 3 g/kg mixed extract] or reduced glutathione for injection(RGI,300 mg/kg) for 30 min before exposure to lipopolysaccharide(LPS,16 mg/kg). After intraperitoneal LPS stimulation for 90 min or 6 h,the mice were sacrificed by ether anaesthesia,and serum and liver samples were collected. Histological analysis(H&E) and terminal deoxynucleotidyl transferase-mediated d UTP nick end-labelling(TUNEL) staining were performed. Alanine transferase(ALT),aspartate transaminase(AST),total bilirubin(TBil),glutathione-S-transferase(GST),malondialdehyde(MDA),tumour necrosis factor(TNF)-α,interleukin(IL)-6,and caspase-3 levels were measured. Bax,Bcl-2,P-IκBα,IκBα,P-NF-κB p65,and NF-κB p65 protein levels were determined by Western blot. TNF-α,IL-6,caspase-3,Bax and Bcl-2 m RNA expression was measured by real-time reverse transcription-polymerase chain reaction(RT-PCR).RESULTS:Hematoxylin-eosin staining and TUNEL results suggested that DHI(3 g/kg) treatment alleviated inflammatory and apoptotic(P < 0.01) injury in the liver of mice. DHI treatment dose-dependently blunted the abnormal changes in biochemical parameters such as ALT(72.53 ± 2.83 for 3 g/kg,P < 0.01),AST(76.97 ± 5.00 for 3 g/kg,P < 0.01),TBil(1.17 ± 0.10 for 3 g/kg,P < 0.01),MDA(0.81 ± 0.36 for 3 g/kg,P < 0.01),and GST(358.86 ± 12.09 for 3 g/kg,P < 0.01). Moreover,DHI(3 g/kg) remarkably decreased LPS-induced protein expression of TNF-α(340.55 ± 10.18 for 3 g/kg,P < 0.01),IL-6(261.34 ± 10.18 for 3 g/kg,P < 0.01),and enzyme activity of caspase-3(0.93 ± 0.029 for 3 g/kg,P < 0.01). The LPS-induced m RNA expression of TNF-α,IL-6 and caspase-3 was also decreased by DHI. Western blot analysis revealed that DHI antagonised LPS-stimulated decrease of Bcl-2 and increase of Bax protein expression. Furthermore,DHI inhibited LPS-induced IκBα and NF-κB p65 phosphorylation.CONCLUSION:DHI may be a multi-function protectant against acute hepatic injury in mice through its antiinflammatory,anti-oxidative and anti-apoptotic activities.
基金The work was financially supported by Special Research Fund for Young Doctors of Qiqihar Academy of Medical Sciences(QMSI2020B-02,QMSI2020B-03)Postdoctoral Science Development Foundation Heilongjiang Province of China(LBH-Q17179).
文摘Based on network pharmacology and molecular docking technology,the similarities and differences of anti-myocardial ischemia mechanism between Salvia miltiorrhiza Bunge(SM)and Carthamus tinctorius L.(CT)were studied.Firstly,based on the traditional Chinese medicine systems pharmacology(TCMSP),the related compounds of SM and CT were obtained,and the potential targets of these compounds were collected by the target fishing method.Genecards database was used to obtain targets related to myocardial ischemia.The cross targets of CT,SM,and myocardial ischemia were then selected,and the protein-protein interaction(PPI)network was constructed based on the STRING database.The cross targets were imported into the Metascape database for Gene Oncology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.Cytoscape software was used to build the topological network diagram of the drug-compound-target path.Finally,the binding ability of the active ingredient and the key target was verified by molecular docking.65 active ingredients and 38 potential targets were screened from SM,and 22 active ingredients and 58 potential targets were excavated from CT.Important targets common to SM and CT were TNF,IL6,VEGFA,AKT1,etc.The common enrichment pathways involved are fluid shear stress and atherosclerosis,IL-17 signaling pathway,pathways in cancer,and toxoplasmosis.The findings suggested that the two traditional Chinese medicines exerted the effect of myocardial ischemia through the characteristics of multiple targets,multiple pathways,and multiple compounds.
文摘目的基于网络药理学筛选丹参-红花药对配伍抗缺血性脑卒中的成分靶点、作用靶点,并对大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)大鼠模型进行相关作用靶点的实验验证。方法①通过系统药理学平台(Traditional Chinese Medicine Systems Pharmacology,TCMSP)、基因名片(GeneCards)数据库筛选丹参-红花的有效成分、成分靶点及缺血性脑卒中的相关靶点;运用STRING数据库构建蛋白互作网络图(protein protein interaction network,PPI),并利用生物信息分析软件(Cytoscape3.8.0)构建成分靶点互作网络;利用David在线分析工具对目标基因进行基因本体论(gene ontology,GO)富集分析和基因的京都百科全书和基因组(kyoto encyclopedia of genes and genomes,KEGG)通路分析。②采用改良MCAO法制备缺血性脑卒中大鼠模型,采用免疫组化法、实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠脑组织中NLR家族Pyrin域蛋白3(NLRP3)炎症小体和核转录因子-B(NF-kB)p65(NFkBp65)蛋白的阳性表达,探讨丹红配伍抗脑缺血损伤炎症反应的作用机制。结果①丹参-红花药对中筛选出87个有效成分,253个靶点,缺血性脑卒中疾病靶点1448个,药物与疾病相关靶点161个,通过GO分析得出730个生物学过程(biological process,BP),81个细胞组分(cellular component,CC),128个分子功能(molecular function,MF),通过KEGG分析得出127条信号通路。②免疫组化法和Real-time PCR测定结果显示,与假手术组比较,模型组大鼠脑组织中NLRP3炎症小体和NFkBp65蛋白表达明显增加(P<0.01),与模型组比较,丹红配伍组和尼莫地平组NLRP3炎症小体和NFkBp65蛋白表达明显降低(P<0.01)。结论丹红有效组分配伍能够通过NF-κB信号通路下调NLRP3炎症小体的释放,阻断炎症损伤环节,最终达到抗炎损伤的作用。
