OBJECTIVE To investigate the regulatory effects of icariin(ICA)on cardiac micro⁃vascular endothelial cells(CMEC)after oxygenglucose deprivation reperfusion(OGD/R)injury.METHODS CMEC were subjected to OGD/R treatment t...OBJECTIVE To investigate the regulatory effects of icariin(ICA)on cardiac micro⁃vascular endothelial cells(CMEC)after oxygenglucose deprivation reperfusion(OGD/R)injury.METHODS CMEC were subjected to OGD/R treatment to construct a myocardial ischemiareperfusion model,and were divided into normal,model,low(10μmol·L^(-1)),medium(20μmol·L^(-1))and high(40μmol·L^(-1))ICA group,and high ICA+inhibitor group(40μmol·L^(-1)+20 nmol·L^(-1)).CCK-8 assay was used to assess the protective ability of ICA against CMEC,and cell migration assay and tube-formation assay were used to detect the migration and generation ability of CMEC.The TCMSP database,Swiss-Target database and literature mining methods were used to col⁃lect ICA-related targets,the GeneCards data⁃base was used to collect target genes related to myocardial ischemia/reperfusion,and Cytoscape 3.8.0 software was used to construct a"drug-tar⁃get-disease"network.The potential targets were imported into STRING 11.5 database to obtain the PPI network.GO and KEGG enrichment analyses were performed on the potential targets using the DAVID database.Molecular docking was performed using AutoDock-vina 1.1.2 soft⁃ware.Western blot detected the expression of related proteins.RESULTS After CMEC was subjected to OGD/R treatment,ICA had a protec⁃tive effect at 10^(-1)60μmol·L^(-1);the results of the cell migration assay showed that each group of ICA could promote the migratory effect of CMEC(P<0.01,P<0.01);and the results of tube-for⁃mation assay showed that each group of ICA could significantly promote the generation of branches(P<0.01)and the capillary length exten⁃sion(P<0.05).Network pharmacology collected a total of 23 ICA action targets,1500 disease tar⁃gets and 12 key targets.GO function enrichment analysis found 85 results.KEGG pathway enrich⁃ment analysis found 53 results,involving AGERAGE signaling pathway,sphingolipid signaling pathway and VEGF signaling pathway.Molecu⁃lar docking results showed that ICA had better binding with core targets PRKCB,PRKCA and PTGS2.Western blot results showed that ICA could regulate the expression of PRKCB,PRKCA and PTGS2 proteins.The results of cell migra⁃tion assay,tube-formation assay and protein expression were reversed after addition of PKC inhibitor.CONCLUSION The potential mecha⁃nism of action of ICA against myocardial isch⁃emia-reperfusion injury may be related to the reg⁃ulation of processes such as CMEC migration and angiogenesis,and it functions through the key target gene PKC.展开更多
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 isolate and identify the undescribed compounds from the fruits of Cinnamomum migao and evaluate its nitric oxide inhibition potential.Methods:The chromatographic techniques of silica gel,Sephadex,and HPLC...Objective:To isolate and identify the undescribed compounds from the fruits of Cinnamomum migao and evaluate its nitric oxide inhibition potential.Methods:The chromatographic techniques of silica gel,Sephadex,and HPLC were used for isolation and purification of the compounds,while HR-ESI-MS,1D NMR,2D NMR,ECD,and X-ray diffraction techniques were used to characterize and confirm the isolated compounds.Moreover,the antiinflammatory activity of the isolated compounds was carried out to check inhibitory potential against the production of nitric oxide with RAW264.7 cells stimulated by LPS.Results:Camganoid A(1),a novel sesquiterpene possessing an unprecedented skeleton,and camganoid B(2),containing a unique eight-membered sesquiterpene moiety with a new carbon skeleton,were isolated and identified from the fruits of C.migao.The absolute configurations of 1 and 2 were confirmed by single crystal X-ray diffraction and electronic circular dichroism(ECD)calculations.Among these compounds,compound 1 exhibited potent inhibitory activity against the production of nitric oxide with IC50value of 4.59μmol/L in RAW264.7 cells stimulated by LPS.Conclusion:The isolation of two new skeletons from the fruits part of C.migao possessed unique skeletons which have not been reported before.展开更多
Objective To establish a reverse-phase liquid chromatography method for the determination of seven alkaloids (magnoflorine, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, and berberine) in Fufang Zhen...Objective To establish a reverse-phase liquid chromatography method for the determination of seven alkaloids (magnoflorine, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, and berberine) in Fufang Zhenzhu Tiaozhi Capsule. Methods Chromatography was performed on a Dionex Acclaim C 18 column (250 mm × 4.6 mm, 5.0 μm) at 30 ℃ .The mobile phase was composed of acetonitrile-potassium dihydrogen phosphate solution (0.015 mol/L, 40:60, including 1.7 g/L sodium dodecyl sulfate and phosphoric acid used to regulate pH value to 3.0), with a flow rate of 1.0 mL/min. The detection wavelength was 270 nm. Results The calibration curves of magnoflorine, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, and berberine were linear in the range of 1.07- 10.65, 0.78-7.55, 0.75-7.50, 1.60-15.95, 2.69-26.85, 2.31-23.10, and 6.04-60.40 mg/mL. The average recoveries of magnoflorine, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, and berberine were 101.0%, 101.2%, 100.1%, 100.0%, 100.1%, 101.1%, and 99.7%, respectively. Conclusion The method could be used for the quantitative determination of the preparation.展开更多
Objective:To explore the protective effect of Huoxin Pill(HXP)on acute myocardial ischemia-reperfusion(MIRI)injury in rats.Methods:Seventy-five adult SD rats were divided into the sham-operated group,model group,posit...Objective:To explore the protective effect of Huoxin Pill(HXP)on acute myocardial ischemia-reperfusion(MIRI)injury in rats.Methods:Seventy-five adult SD rats were divided into the sham-operated group,model group,positive drug group(diltiazem hydrochloride,DH),high dose group(24 mg/kg,HXP-H)and low dose group(12 mg/kg,HXP-L)of Huoxin Pill(n=15 for every group)according to the complete randomization method.After 1 week of intragastric administration,the left anterior descending coronary artery of the rat's heart was ligated for 45 min and reperfused for 3 h.Serum was separated and the levels of creatine kinase(CK),creatine kinase isoenzyme(CK-MB)and lactate dehydrogenase(LDH),superoxide dismutase(SOD),and malondialdehyde(MDA),hypersensitive C-reactive protein(hs-CRP)and interleukin-1β(IL-1β)were measured.Myocardial ischemia rate,myocardial infarction rate and myocardial no-reflow rate were determined by staining with Evans blue and 2,3,5-triphenyltetrazolium chloride(TTC).Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMAN)databases were used to screen for possible active compounds of HXP and their potential therapeutic targets;the results of anti-inflammatory genes associated with MIRI were obtained from GeneC ards,Drugbank,Online Mendelian Inheritance in Man(OMIM),and Therapeutic Target Datebase(TTD)databases was performed;Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment were used to analyze the intersected targets;molecular docking was performed using AutoD ock Tools.Western blot was used to detect the protein expression of Toll-like receptor 4(TLR4)/nuclear factor kappa-B(NFκB)/NOD-like receptor protein 3(NLRP3).Results:Compared with the model group,all doses of HXP significantly reduced the levels of LDH,CK and CK-MB(P<0.05,P<0.01);HXP significantly increased serum activity of SOD(P<0.05,P<0.01);all doses of HXP significantly reduced the levels of hs-CRP and IL-1β(P<0.05,P<0.01)and the myocardial infarction rate and myocardial no-reflow rate(P<0.01).GO enrichment analysis mainly involved positive regulation of gene expression,extracellular space and identical protein binding,KEGG pathway enrichment mainly involved PI3K-Akt signaling pathway and lipid and atherosclerosis.Molecular docking results showed that kaempferol and luteolin had a better affinity with TLR4,NFκB and NLRP3 molecules.The protein expressions of TLR4,NFκB and NLRP3 were reduced in the HXP group(P<0.01).Conclusions:HXP has a significant protective effect on myocardial ischemia-reperfusion injury in rats,and its effect may be related to the inhibition of redox response and reduction of the inflammatory response by inhibiting the TLR4/NFκB/NLRP3 signaling pathway.展开更多
Objective: To isolate the phenolic amides from the dried bulbs of Allium chinense and investigate their myocardium protective activities.Methods: The chemical constituents were isolated and purified by combining with ...Objective: To isolate the phenolic amides from the dried bulbs of Allium chinense and investigate their myocardium protective activities.Methods: The chemical constituents were isolated and purified by combining with silica gel column,Sephadex LH-20 column, HPLC and other chromatography techniques. Their structures were elucidated by NMR techniques and mass spectrometry. The isolated compounds were evaluated to determine their protective effect for myocardium cells in vitro.Results: Two new phenolic amides, namely, alichinemide Ⅰ(1) and alichinemide Ⅱ(2), and six konwn amides were isolated from the dried bulbs of A. chinense. The structures of compounds 3–8 were identified as 3-indolcarbaldehyde(3), 1-(2-aminophenyl)urea(4), 2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylic acid(5), N-trans-feruloyltyramine(6), N-trans-p-coumaroyltyramine(7), and N-(3,4-dimethoxyphenethyl) acetamide(8). Compound 3(50 μmol/L) showed significant inhibitory effect on the damage of H9c2 myocardial cells induced by H2O2in vitro.Conclusion: Compounds 1 and 2 were new phenolic amides. Compound 3 could be one of the potential myocardium protective constituents of A. chinense.展开更多
基金National Natural Science Foundation of China(82030124)National Natural Science Foundation of China(82174015)Science and Technology Innovation Project of China Academy of Traditional Chinese Medicine(CI2021A04609)。
文摘OBJECTIVE To investigate the regulatory effects of icariin(ICA)on cardiac micro⁃vascular endothelial cells(CMEC)after oxygenglucose deprivation reperfusion(OGD/R)injury.METHODS CMEC were subjected to OGD/R treatment to construct a myocardial ischemiareperfusion model,and were divided into normal,model,low(10μmol·L^(-1)),medium(20μmol·L^(-1))and high(40μmol·L^(-1))ICA group,and high ICA+inhibitor group(40μmol·L^(-1)+20 nmol·L^(-1)).CCK-8 assay was used to assess the protective ability of ICA against CMEC,and cell migration assay and tube-formation assay were used to detect the migration and generation ability of CMEC.The TCMSP database,Swiss-Target database and literature mining methods were used to col⁃lect ICA-related targets,the GeneCards data⁃base was used to collect target genes related to myocardial ischemia/reperfusion,and Cytoscape 3.8.0 software was used to construct a"drug-tar⁃get-disease"network.The potential targets were imported into STRING 11.5 database to obtain the PPI network.GO and KEGG enrichment analyses were performed on the potential targets using the DAVID database.Molecular docking was performed using AutoDock-vina 1.1.2 soft⁃ware.Western blot detected the expression of related proteins.RESULTS After CMEC was subjected to OGD/R treatment,ICA had a protec⁃tive effect at 10^(-1)60μmol·L^(-1);the results of the cell migration assay showed that each group of ICA could promote the migratory effect of CMEC(P<0.01,P<0.01);and the results of tube-for⁃mation assay showed that each group of ICA could significantly promote the generation of branches(P<0.01)and the capillary length exten⁃sion(P<0.05).Network pharmacology collected a total of 23 ICA action targets,1500 disease tar⁃gets and 12 key targets.GO function enrichment analysis found 85 results.KEGG pathway enrich⁃ment analysis found 53 results,involving AGERAGE signaling pathway,sphingolipid signaling pathway and VEGF signaling pathway.Molecu⁃lar docking results showed that ICA had better binding with core targets PRKCB,PRKCA and PTGS2.Western blot results showed that ICA could regulate the expression of PRKCB,PRKCA and PTGS2 proteins.The results of cell migra⁃tion assay,tube-formation assay and protein expression were reversed after addition of PKC inhibitor.CONCLUSION The potential mecha⁃nism of action of ICA against myocardial isch⁃emia-reperfusion injury may be related to the reg⁃ulation of processes such as CMEC migration and angiogenesis,and it functions through the key target gene PKC.
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 NSFC(81973191)Natural Science Foundation of Shanghai(19ZR1428100)+1 种基金Project supported by the Department of Science and Technology of Guizhou(Grant No.[2018]2831)the Horizontal Research Project of Shanghai Jiao Tong University(SA1700111,SA1700118)。
文摘Objective:To isolate and identify the undescribed compounds from the fruits of Cinnamomum migao and evaluate its nitric oxide inhibition potential.Methods:The chromatographic techniques of silica gel,Sephadex,and HPLC were used for isolation and purification of the compounds,while HR-ESI-MS,1D NMR,2D NMR,ECD,and X-ray diffraction techniques were used to characterize and confirm the isolated compounds.Moreover,the antiinflammatory activity of the isolated compounds was carried out to check inhibitory potential against the production of nitric oxide with RAW264.7 cells stimulated by LPS.Results:Camganoid A(1),a novel sesquiterpene possessing an unprecedented skeleton,and camganoid B(2),containing a unique eight-membered sesquiterpene moiety with a new carbon skeleton,were isolated and identified from the fruits of C.migao.The absolute configurations of 1 and 2 were confirmed by single crystal X-ray diffraction and electronic circular dichroism(ECD)calculations.Among these compounds,compound 1 exhibited potent inhibitory activity against the production of nitric oxide with IC50value of 4.59μmol/L in RAW264.7 cells stimulated by LPS.Conclusion:The isolation of two new skeletons from the fruits part of C.migao possessed unique skeletons which have not been reported before.
基金Cooperation Project in Industry, Education and Research of Guangdong Province and Ministry of Education of China (2009B090300349)a Cooperation Project of National Science Foundation of Guangdong Province (10351022401000000)
文摘Objective To establish a reverse-phase liquid chromatography method for the determination of seven alkaloids (magnoflorine, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, and berberine) in Fufang Zhenzhu Tiaozhi Capsule. Methods Chromatography was performed on a Dionex Acclaim C 18 column (250 mm × 4.6 mm, 5.0 μm) at 30 ℃ .The mobile phase was composed of acetonitrile-potassium dihydrogen phosphate solution (0.015 mol/L, 40:60, including 1.7 g/L sodium dodecyl sulfate and phosphoric acid used to regulate pH value to 3.0), with a flow rate of 1.0 mL/min. The detection wavelength was 270 nm. Results The calibration curves of magnoflorine, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, and berberine were linear in the range of 1.07- 10.65, 0.78-7.55, 0.75-7.50, 1.60-15.95, 2.69-26.85, 2.31-23.10, and 6.04-60.40 mg/mL. The average recoveries of magnoflorine, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, and berberine were 101.0%, 101.2%, 100.1%, 100.0%, 100.1%, 101.1%, and 99.7%, respectively. Conclusion The method could be used for the quantitative determination of the preparation.
基金Supported by the National Natural Science Foundation of China (No.82174015 and No.82030124)Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences (No.CI2021A04609)。
文摘Objective:To explore the protective effect of Huoxin Pill(HXP)on acute myocardial ischemia-reperfusion(MIRI)injury in rats.Methods:Seventy-five adult SD rats were divided into the sham-operated group,model group,positive drug group(diltiazem hydrochloride,DH),high dose group(24 mg/kg,HXP-H)and low dose group(12 mg/kg,HXP-L)of Huoxin Pill(n=15 for every group)according to the complete randomization method.After 1 week of intragastric administration,the left anterior descending coronary artery of the rat's heart was ligated for 45 min and reperfused for 3 h.Serum was separated and the levels of creatine kinase(CK),creatine kinase isoenzyme(CK-MB)and lactate dehydrogenase(LDH),superoxide dismutase(SOD),and malondialdehyde(MDA),hypersensitive C-reactive protein(hs-CRP)and interleukin-1β(IL-1β)were measured.Myocardial ischemia rate,myocardial infarction rate and myocardial no-reflow rate were determined by staining with Evans blue and 2,3,5-triphenyltetrazolium chloride(TTC).Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMAN)databases were used to screen for possible active compounds of HXP and their potential therapeutic targets;the results of anti-inflammatory genes associated with MIRI were obtained from GeneC ards,Drugbank,Online Mendelian Inheritance in Man(OMIM),and Therapeutic Target Datebase(TTD)databases was performed;Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment were used to analyze the intersected targets;molecular docking was performed using AutoD ock Tools.Western blot was used to detect the protein expression of Toll-like receptor 4(TLR4)/nuclear factor kappa-B(NFκB)/NOD-like receptor protein 3(NLRP3).Results:Compared with the model group,all doses of HXP significantly reduced the levels of LDH,CK and CK-MB(P<0.05,P<0.01);HXP significantly increased serum activity of SOD(P<0.05,P<0.01);all doses of HXP significantly reduced the levels of hs-CRP and IL-1β(P<0.05,P<0.01)and the myocardial infarction rate and myocardial no-reflow rate(P<0.01).GO enrichment analysis mainly involved positive regulation of gene expression,extracellular space and identical protein binding,KEGG pathway enrichment mainly involved PI3K-Akt signaling pathway and lipid and atherosclerosis.Molecular docking results showed that kaempferol and luteolin had a better affinity with TLR4,NFκB and NLRP3 molecules.The protein expressions of TLR4,NFκB and NLRP3 were reduced in the HXP group(P<0.01).Conclusions:HXP has a significant protective effect on myocardial ischemia-reperfusion injury in rats,and its effect may be related to the inhibition of redox response and reduction of the inflammatory response by inhibiting the TLR4/NFκB/NLRP3 signaling pathway.
基金supported by Natural Science Foundation of China (No. 81973191)Shanghai Natural Science Fund (No. 19ZR1428100)+1 种基金project supported by the Department of Science and Technology of Guizhou (No. [2018] 2831)the Horizontal Research Project of Shanghai Jiao Tong University (Nos. SA1700111, SA1700118)。
文摘Objective: To isolate the phenolic amides from the dried bulbs of Allium chinense and investigate their myocardium protective activities.Methods: The chemical constituents were isolated and purified by combining with silica gel column,Sephadex LH-20 column, HPLC and other chromatography techniques. Their structures were elucidated by NMR techniques and mass spectrometry. The isolated compounds were evaluated to determine their protective effect for myocardium cells in vitro.Results: Two new phenolic amides, namely, alichinemide Ⅰ(1) and alichinemide Ⅱ(2), and six konwn amides were isolated from the dried bulbs of A. chinense. The structures of compounds 3–8 were identified as 3-indolcarbaldehyde(3), 1-(2-aminophenyl)urea(4), 2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylic acid(5), N-trans-feruloyltyramine(6), N-trans-p-coumaroyltyramine(7), and N-(3,4-dimethoxyphenethyl) acetamide(8). Compound 3(50 μmol/L) showed significant inhibitory effect on the damage of H9c2 myocardial cells induced by H2O2in vitro.Conclusion: Compounds 1 and 2 were new phenolic amides. Compound 3 could be one of the potential myocardium protective constituents of A. chinense.
