OBJECTIVE:To evaluate the efficacy of Renshen Sanqi Chuanxiong formula(RSCF)for preventing vascular aging,and to investigate the possible molecular mechanism underlying the actions of RSCF.METHODS:Potentially active c...OBJECTIVE:To evaluate the efficacy of Renshen Sanqi Chuanxiong formula(RSCF)for preventing vascular aging,and to investigate the possible molecular mechanism underlying the actions of RSCF.METHODS:Potentially active components and their relatively direct targets were identified by combining drug-likeness(DL)screening using a target identification process.Vascular aging-associated targets for RSCF were obtained by selecting common genes not only from potential targets but also from human vascular aging-associated genes.Cytoscape 3.2.1 software was employed to visualize the complex compound-target and target-function networks.Biological process and molecular function were assessed,and the Kyoto Encyclopedia of Genes and Genomes and pathway enrichment analyses were performed using Clue GO.Pathways directly associated with vascular aging were integrated into a"vascular aging-related"pathway.RESULTS:Altogether,122 potentially active components of RSCF were identified through DL screening,and their corresponding 692 direct targets were retrieved via target prediction and identification.We identified 49 vascular aging-associated targets for RSCF by overlapping the 692 potential targets with 146 human vascular aging-associated genes.The results from the compound-target network indicated that most components acted on common targets and displayed synergistic action,which showed that the magnifying effects of RSCF were based on these common targets.The target-function network revealed that each target was involved in multiple function modules,suggesting that RSCF was multi-functional during treatment of vascular aging.The results of the Clue GO analysis indicated that most of the targets were associated with the hypoxia-inducible factor 1(HIF-1)signaling pathway.The results from the pathway analysis also indicated that an integrative vascular aging-related pathway mainly included an angiogenesis regulation module,cell-survival module,and oxidative stress-resistance module.CONCLUSION:Our results suggested that many components act synergistically on common targets to delay vascular aging,and each target is involved in multiple functional modules.The Clue GO analysis indicated that most of the targets were connected to the HIF-1 signaling pathway,FOXO signaling pathway,and thyroid hormone signaling pathway.展开更多
OBJECTIVE: To investigate the effects of extracts from Renshen(Radix Ginseng), Sanqi(Radix Notoginseng), and Chuanxiong(Rhizoma Chuanxiong) on the endothelial actin cytoskeleton in senescent human cardiac microvascula...OBJECTIVE: To investigate the effects of extracts from Renshen(Radix Ginseng), Sanqi(Radix Notoginseng), and Chuanxiong(Rhizoma Chuanxiong) on the endothelial actin cytoskeleton in senescent human cardiac microvascular endothelial cells(HCMECs), and to propose the possible mechanism underlying the actions.METHODS: Lentiviral mediated RNA interference was applied to a replicative senescent HCMEC model by knocking down heat shock protein 27(HSP27)gene. Cells were treated with extracts from Renshen(Radix Ginseng), Sanqi(Radix Notoginseng), and Chuanxiong(Rhizoma Chuanxiong) at final concentrations of 50, 100, and 200 mg/L, respectively and with 10 μM resveratrol for 48 h. Untreated cells were used as controls. Senescence was detected by senescence β-galactosidase staining and cell proliferation was analyzed by cell counting kit-8 assays.Secreted nitric oxide levels were detected by nitrate reductase. Morphological changes of F-actin and G-actin were observed by laser scanning confocal microscopy. Protein and gene expression of Factin and HSP27 was detected by western blotting.RESULTS: Compared with the control group, the proportion of senescent HSP27 shRNA cells treated with the extracts was decreased and their proliferation was increased. In the extract intervention group, F-actin around the cell periphery became irregular and jagged fractures formed gradually and then dissipated. Moreover, some dynamic actin stress fiber filaments appeared. The G-actin stretched to the cell periphery and punctate staining was scattered in the cytoplasm. In addition, the mean optical density value of F/G-actin was decreased significantly and the protein expression of F-actin was downregulated.CONCLUSION: The extracts delayed microvascular endothelial cell senescence by downregulating the expression of F-actin through HSP27.展开更多
基金Supported by the National Nature Science Foundation of China(the Mechamism research of Vascular Aging Induced by High Glucose from AMPK/m TOR Pathway and the Interfering Effects of Yiqi Huoxue Chinese Herbal Medicine,No.81673822,the Molecular Mechamism Research of Yiqi Huoxue Fang Postponing Vascular Endothelial Cell Senescence by SIRT1-autophagy Pathway,No.81503448)the Independent Topic Program of China Academy of Chinese Medical Sciences(Effect of Intestinal Microflora on High Glucose-induced Vascular Aging and Internention of Extracts from Radix Ginseng Radix Notoginseng and Rhizoma Chanxiong,No.ZZ2017011)
文摘OBJECTIVE:To evaluate the efficacy of Renshen Sanqi Chuanxiong formula(RSCF)for preventing vascular aging,and to investigate the possible molecular mechanism underlying the actions of RSCF.METHODS:Potentially active components and their relatively direct targets were identified by combining drug-likeness(DL)screening using a target identification process.Vascular aging-associated targets for RSCF were obtained by selecting common genes not only from potential targets but also from human vascular aging-associated genes.Cytoscape 3.2.1 software was employed to visualize the complex compound-target and target-function networks.Biological process and molecular function were assessed,and the Kyoto Encyclopedia of Genes and Genomes and pathway enrichment analyses were performed using Clue GO.Pathways directly associated with vascular aging were integrated into a"vascular aging-related"pathway.RESULTS:Altogether,122 potentially active components of RSCF were identified through DL screening,and their corresponding 692 direct targets were retrieved via target prediction and identification.We identified 49 vascular aging-associated targets for RSCF by overlapping the 692 potential targets with 146 human vascular aging-associated genes.The results from the compound-target network indicated that most components acted on common targets and displayed synergistic action,which showed that the magnifying effects of RSCF were based on these common targets.The target-function network revealed that each target was involved in multiple function modules,suggesting that RSCF was multi-functional during treatment of vascular aging.The results of the Clue GO analysis indicated that most of the targets were associated with the hypoxia-inducible factor 1(HIF-1)signaling pathway.The results from the pathway analysis also indicated that an integrative vascular aging-related pathway mainly included an angiogenesis regulation module,cell-survival module,and oxidative stress-resistance module.CONCLUSION:Our results suggested that many components act synergistically on common targets to delay vascular aging,and each target is involved in multiple functional modules.The Clue GO analysis indicated that most of the targets were connected to the HIF-1 signaling pathway,FOXO signaling pathway,and thyroid hormone signaling pathway.
基金Supported by the National Natural Science Foundation of China(the Study on Intervention and Mechanism of Chinese Herbal Medicines for Nourishing Qi and Resolving Blood Stasis on Cytoskeletal Protein of Vascular Aging(No.81273976)the Mechanism of Exploration about Formula for Reinforcing Qi,Warming Yang,Activating Blood and Promoting Diuresis in the Prevention and Treatment of Heart Failure through mT ORC1-Mitophagy Pathway(No.8170-3865)+2 种基金Independent Research Program of the China Academy of Chinese Medical Sciences(the Study on Mechanism of Chinese Herbal Medicines for Nourishing Qi and Resolving Blood Stasis Delaying Vascular Aging(No.zz2013002)the Mechanism of Exploration about Formula for Reinforcing Qi,Warming Yang,Activating Blood and Promoting Diuresis in the Prevention and Treatment of Heart Failure through Nrf2-iron Death Pathway(No.zz13-YQ-034)Beijing Science and Technology Bureau(Clinical study about Formula for Reinforcing Qi,Warming Yang,Activating Blood and Promoting Diuresis to Improve the Survival Status of Patients with Chronic Heart Failure(No.Z171100001017108)。
文摘OBJECTIVE: To investigate the effects of extracts from Renshen(Radix Ginseng), Sanqi(Radix Notoginseng), and Chuanxiong(Rhizoma Chuanxiong) on the endothelial actin cytoskeleton in senescent human cardiac microvascular endothelial cells(HCMECs), and to propose the possible mechanism underlying the actions.METHODS: Lentiviral mediated RNA interference was applied to a replicative senescent HCMEC model by knocking down heat shock protein 27(HSP27)gene. Cells were treated with extracts from Renshen(Radix Ginseng), Sanqi(Radix Notoginseng), and Chuanxiong(Rhizoma Chuanxiong) at final concentrations of 50, 100, and 200 mg/L, respectively and with 10 μM resveratrol for 48 h. Untreated cells were used as controls. Senescence was detected by senescence β-galactosidase staining and cell proliferation was analyzed by cell counting kit-8 assays.Secreted nitric oxide levels were detected by nitrate reductase. Morphological changes of F-actin and G-actin were observed by laser scanning confocal microscopy. Protein and gene expression of Factin and HSP27 was detected by western blotting.RESULTS: Compared with the control group, the proportion of senescent HSP27 shRNA cells treated with the extracts was decreased and their proliferation was increased. In the extract intervention group, F-actin around the cell periphery became irregular and jagged fractures formed gradually and then dissipated. Moreover, some dynamic actin stress fiber filaments appeared. The G-actin stretched to the cell periphery and punctate staining was scattered in the cytoplasm. In addition, the mean optical density value of F/G-actin was decreased significantly and the protein expression of F-actin was downregulated.CONCLUSION: The extracts delayed microvascular endothelial cell senescence by downregulating the expression of F-actin through HSP27.
