The purpose of this study was to establish a high-performance liquid chromatography (HPLC) method for the simultaneous determination of sodium danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B, a...The purpose of this study was to establish a high-performance liquid chromatography (HPLC) method for the simultaneous determination of sodium danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B, and 4-coumaric acid in Danhong injection. The chromatographic method employed was as follows: the column was a Welch Ultimate XB-C18 column (250 mm × 4.6 mm, 10 μm), the mobile phase was a gradient elution of 0.4% formic acid aqueous solution (A) and acetonitrile (B), the detection wavelengths were 280 nm for sodium danshensu, protocatechuic aldehyde, and salvianolic acid B and 326 nm for 4-coumaric acid and rosmarinic acid, the sample volume was 10 μL, the flow rate was 1.0 mL/min, and the column temperature was 35°C. This method can realize the separation and determination of sodium danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B, and 4-coumaric acid within 50 minutes. The linear relationships of the five peak areas and their concentrations are good (R<sup>2</sup>> 0.9997). The precision RSD values are all less than 1.0%. The reproducibility RSD values are all less than 1.3%. The stability RSD values are all less than 2.2%. The recovery values ranged from 92.4% to 99.4%. This method is simple, accurate, and reproducible. It can be used for the determination of sodium danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B, and 4-coumaric acid in Danhong injection.展开更多
Objective:To investigate the involvement of endothelial cells(ECs)-derived exosomes in the antiapoptotic effect of Danhong Injection(DHI)and the mechanism of DHI-induced exosomal protection against postinfarction myoc...Objective:To investigate the involvement of endothelial cells(ECs)-derived exosomes in the antiapoptotic effect of Danhong Injection(DHI)and the mechanism of DHI-induced exosomal protection against postinfarction myocardial apoptosis.Methods:A mouse permanent myocardial infarction(MI)model was established,followed by a 14-day daily treatment with DHI,DHI plus GW4869(an exosomal inhibitor),or saline.Phosphatebuffered saline(PBS)-induced ECs-derived exosomes were isolated,analyzed by miRNA microarray and validated by droplet digital polymerase chain reaction(ddP CR).The exosomes induced by DHI(DHI-exo),PBS(PBS-exo),or DHI+GW4869(GW-exo)were isolated and injected into the peri-infarct zone following MI.The protective effects of DHI and DHI-exo on MI hearts were measured by echocardiography,Masson's trichrome staining,and TUNEL apoptosis assay.The Western blotting and quantitative reverse transcription PCR(qR T-PCR)were used to evaluate the expression levels of miR-125b/p53-mediated pathway components,including miR-125b,p53,Bak,Bax,and caspase-3 activities.Results:DHI significantly improved cardiac function and reduced infarct size in MI mice(P<0.01),which was abolished by the GW4869 intervention.DHI promoted the exosomal secretion in ECs(P<0.01).According to the results of exosomal miRNA microarray assay,30 differentially expressed miRNAs in the DHI-exo were identified(28 up-regulated miRNAs and 2 down-regulated miRNAs).Among them,DHI significantly elevated miR-125b level in DHI-exo and DHI-treated ECs,a recognized apoptotic inhibitor impeding p53 signaling(P<0.05).Remarkably,treatment with DHI and DHI-exo attenuated apoptosis,elevated miR-125b expression level,inhibited capsase-3 activity,and down-regulated the expression levels of proapoptotic effectors(p53,Bak,and Bax)in post-MI hearts,whereas these effects were blocked by GW4869(P<0.05 or P<0.01).Conclusion:DHI and DHI-induced exosomes inhibited apoptosis,promoted the miR-125b expression level,and regulated the p53 apoptotic pathway in post-infarction myocardium.展开更多
To characterize and identify multiple constituents in Danhong injection(DHI), a fast ultra-high performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight tandem mass spectrometry(U...To characterize and identify multiple constituents in Danhong injection(DHI), a fast ultra-high performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight tandem mass spectrometry(UHPLC-ESI-QTOF/MS) method was established and validated in the present study. A total of 63 compounds, including 33 phenolic acids, 2 C-glycosyl quinochalcones, 6 flavonoid O-glycosides, 4 iridoid glycosides, 6 organic acids, 5 amino acids, and 3 nucleosides, were identified or tentatively characterized. In conclusion, the UHPLC-ESI-QTOF/MS method is useful and efficient for in-depth structural elucidation of chemical compounds in complex matrices of herbal medicines such as DHI.展开更多
Objective:To explore the effectiveness of Danhong Injection(丹红注射液)on improving microcirculatory injury after percutaneous coronary intervention(PCI)in patients with coronary heart disease(CHD).Methods:A randomize...Objective:To explore the effectiveness of Danhong Injection(丹红注射液)on improving microcirculatory injury after percutaneous coronary intervention(PCI)in patients with coronary heart disease(CHD).Methods:A randomized controlled trial was conducted and 90 patients were enrolled.A random sequence was generated using statistical analysis software.Patients with microcirculatory injuries after PCI were randomly divided into 3 groups for treatment(30 subjects in each group):Danhong Injection group:after PCI,Danghong Injections were given with intravenous administration with 40 mL twice a day for a week;statins intensive group:after PCI,atorvastatin calcium tablets were given oral medication with 80 mg once,and then atorvastatin 40 mg daily for 1 week;the control group:after PCI,atorvastatin calcium tablets were given oral medication with 10–20 mg daily for 1 week.The index of microcirculation resistance(IMR)was used to assess microcirculatory injury during PCI.The IMR of the target vessel was reexamined after 1 week of drug treatment.Results:After one week's drug treatment,IMR was significantly decreased in both statins intensive group and Danhong Injection group compared with the control group(P<0.01),but no difference was found between statins intensive group and Danhong injection group(14.03±2.54 vs.16.03±5.72 U,P=0.080).Conclusions:The efficacy of Danhong Injection is non-inferior to statin.Early use of Danhong Injection after PCI can effectively improve coronary microcirculation injury after PCI.展开更多
Objective:To observe the effect of Danhong injection(DHI)on perioperative metabolomics of unstable angina pectoris(UA)with blood stasis syndrome.Materials and Methods:A prospective,randomized,controlled,and single-bli...Objective:To observe the effect of Danhong injection(DHI)on perioperative metabolomics of unstable angina pectoris(UA)with blood stasis syndrome.Materials and Methods:A prospective,randomized,controlled,and single-blind clinical trial was conducted.Sixty-one UA patients with traditional Chinese medicine blood stasis syndrome undergoing elective percutaneous coronary intervention(PCI)were randomly divided into the Danhong and control groups,and 10 healthy volunteers were included as baseline.The Danhong group received western medicine+DHI treatment,while the control group received western medicine+saline.Nontargeted metabolomics was used to analyze the serum metabolites of healthy volunteers in the Danhong and control groups before and 5 days after PCI.Results:Before treatment,there was no significant difference in serum metabolites between the Danhong and control groups,but there was a significant difference between the two groups and the healthy group.Differential metabolites were clustered mainly in glycerophospholipid,sphingolipid,purine,and amino acid groups,which were generated in their metabolic pathways.After 5 days of PCI,the profiles of serum metabolites were significantly closer between the Danhong-or control-treated groups and that of the healthy group.Furthermore,DHI treatment converted the serum metabolite profile more to that of the healthy group than the control treatment.Conclusion:The beneficial effect of DHI on patients with unstable angina is reflected at the level of serum metabolic biomarkers.展开更多
Objective:The aim of this study is to explore the active ingredients and mechanism of action of danhong injection(DHI)in treating myeloproliferative neoplasms using network pharmacology.Methods:The TCMSP platform and ...Objective:The aim of this study is to explore the active ingredients and mechanism of action of danhong injection(DHI)in treating myeloproliferative neoplasms using network pharmacology.Methods:The TCMSP platform and relevant literature were used to search for the active ingredients and targets of Radix Salviae and Carthami Flos in DHI.Disease targets related to myeloproliferative neoplasms were obtained from the GEO database,GeneCards,and DisGeNET database.The queried component targets were normalized using the UniProt database.Potential targets were identified by constructing protein-protein interactions networks using STRING 11.5 and visualized and analyzed using Cytoscape 3.9.1.GO and KEGG analysis were performed using the Metascape platform,and visualization was done using the built-in plug-in CluoGO or SangerBox platforms with Cytoscape 3.9.1.Results:The active ingredients of DHI for treating myeloproliferative neoplasms mainly consist of flavonoids and o-benzoquinones,including quercetin,luteolin,kaempferol,stigmasterol,tanshinone iia,cryptotanshinone,beta-carotene,2-isopropyl-8-methylphenanthrene-3,4-dione,and neocryptotanshinone ii.The potential targets are JUN,TP53,STAT3,AKT1,MAPK1,RELA,TNF,MAPK14,IL6,and FOS.The relevant signaling pathways involved are mainly TNFαsignaling pathway,PI3K-Akt signaling pathway,apoptosis,IL-17 signaling pathway,cellular senescence,MAPK signaling pathway,p53 signaling pathway,JAK-STAT signaling pathway,and NF-kappa B signaling.Conclusions:DHI acts mainly through flavonoids and o-benzoquinones to treat myeloproliferative neoplasms in a multi-targeted and multi-pathway manner.展开更多
文摘The purpose of this study was to establish a high-performance liquid chromatography (HPLC) method for the simultaneous determination of sodium danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B, and 4-coumaric acid in Danhong injection. The chromatographic method employed was as follows: the column was a Welch Ultimate XB-C18 column (250 mm × 4.6 mm, 10 μm), the mobile phase was a gradient elution of 0.4% formic acid aqueous solution (A) and acetonitrile (B), the detection wavelengths were 280 nm for sodium danshensu, protocatechuic aldehyde, and salvianolic acid B and 326 nm for 4-coumaric acid and rosmarinic acid, the sample volume was 10 μL, the flow rate was 1.0 mL/min, and the column temperature was 35°C. This method can realize the separation and determination of sodium danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B, and 4-coumaric acid within 50 minutes. The linear relationships of the five peak areas and their concentrations are good (R<sup>2</sup>> 0.9997). The precision RSD values are all less than 1.0%. The reproducibility RSD values are all less than 1.3%. The stability RSD values are all less than 2.2%. The recovery values ranged from 92.4% to 99.4%. This method is simple, accurate, and reproducible. It can be used for the determination of sodium danshensu, protocatechuic aldehyde, rosmarinic acid, salvianolic acid B, and 4-coumaric acid in Danhong injection.
基金Supported by National Natural Science Foundation of China (No.82174106 and No.81703850)。
文摘Objective:To investigate the involvement of endothelial cells(ECs)-derived exosomes in the antiapoptotic effect of Danhong Injection(DHI)and the mechanism of DHI-induced exosomal protection against postinfarction myocardial apoptosis.Methods:A mouse permanent myocardial infarction(MI)model was established,followed by a 14-day daily treatment with DHI,DHI plus GW4869(an exosomal inhibitor),or saline.Phosphatebuffered saline(PBS)-induced ECs-derived exosomes were isolated,analyzed by miRNA microarray and validated by droplet digital polymerase chain reaction(ddP CR).The exosomes induced by DHI(DHI-exo),PBS(PBS-exo),or DHI+GW4869(GW-exo)were isolated and injected into the peri-infarct zone following MI.The protective effects of DHI and DHI-exo on MI hearts were measured by echocardiography,Masson's trichrome staining,and TUNEL apoptosis assay.The Western blotting and quantitative reverse transcription PCR(qR T-PCR)were used to evaluate the expression levels of miR-125b/p53-mediated pathway components,including miR-125b,p53,Bak,Bax,and caspase-3 activities.Results:DHI significantly improved cardiac function and reduced infarct size in MI mice(P<0.01),which was abolished by the GW4869 intervention.DHI promoted the exosomal secretion in ECs(P<0.01).According to the results of exosomal miRNA microarray assay,30 differentially expressed miRNAs in the DHI-exo were identified(28 up-regulated miRNAs and 2 down-regulated miRNAs).Among them,DHI significantly elevated miR-125b level in DHI-exo and DHI-treated ECs,a recognized apoptotic inhibitor impeding p53 signaling(P<0.05).Remarkably,treatment with DHI and DHI-exo attenuated apoptosis,elevated miR-125b expression level,inhibited capsase-3 activity,and down-regulated the expression levels of proapoptotic effectors(p53,Bak,and Bax)in post-MI hearts,whereas these effects were blocked by GW4869(P<0.05 or P<0.01).Conclusion:DHI and DHI-induced exosomes inhibited apoptosis,promoted the miR-125b expression level,and regulated the p53 apoptotic pathway in post-infarction myocardium.
基金supported by the National Natural Science Foundation of China(Nos.81130068 and 81503241)a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘To characterize and identify multiple constituents in Danhong injection(DHI), a fast ultra-high performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight tandem mass spectrometry(UHPLC-ESI-QTOF/MS) method was established and validated in the present study. A total of 63 compounds, including 33 phenolic acids, 2 C-glycosyl quinochalcones, 6 flavonoid O-glycosides, 4 iridoid glycosides, 6 organic acids, 5 amino acids, and 3 nucleosides, were identified or tentatively characterized. In conclusion, the UHPLC-ESI-QTOF/MS method is useful and efficient for in-depth structural elucidation of chemical compounds in complex matrices of herbal medicines such as DHI.
基金Supported by Capital Health Development Research Project(No.2014-2-4032)。
文摘Objective:To explore the effectiveness of Danhong Injection(丹红注射液)on improving microcirculatory injury after percutaneous coronary intervention(PCI)in patients with coronary heart disease(CHD).Methods:A randomized controlled trial was conducted and 90 patients were enrolled.A random sequence was generated using statistical analysis software.Patients with microcirculatory injuries after PCI were randomly divided into 3 groups for treatment(30 subjects in each group):Danhong Injection group:after PCI,Danghong Injections were given with intravenous administration with 40 mL twice a day for a week;statins intensive group:after PCI,atorvastatin calcium tablets were given oral medication with 80 mg once,and then atorvastatin 40 mg daily for 1 week;the control group:after PCI,atorvastatin calcium tablets were given oral medication with 10–20 mg daily for 1 week.The index of microcirculation resistance(IMR)was used to assess microcirculatory injury during PCI.The IMR of the target vessel was reexamined after 1 week of drug treatment.Results:After one week's drug treatment,IMR was significantly decreased in both statins intensive group and Danhong Injection group compared with the control group(P<0.01),but no difference was found between statins intensive group and Danhong injection group(14.03±2.54 vs.16.03±5.72 U,P=0.080).Conclusions:The efficacy of Danhong Injection is non-inferior to statin.Early use of Danhong Injection after PCI can effectively improve coronary microcirculation injury after PCI.
文摘Objective:To observe the effect of Danhong injection(DHI)on perioperative metabolomics of unstable angina pectoris(UA)with blood stasis syndrome.Materials and Methods:A prospective,randomized,controlled,and single-blind clinical trial was conducted.Sixty-one UA patients with traditional Chinese medicine blood stasis syndrome undergoing elective percutaneous coronary intervention(PCI)were randomly divided into the Danhong and control groups,and 10 healthy volunteers were included as baseline.The Danhong group received western medicine+DHI treatment,while the control group received western medicine+saline.Nontargeted metabolomics was used to analyze the serum metabolites of healthy volunteers in the Danhong and control groups before and 5 days after PCI.Results:Before treatment,there was no significant difference in serum metabolites between the Danhong and control groups,but there was a significant difference between the two groups and the healthy group.Differential metabolites were clustered mainly in glycerophospholipid,sphingolipid,purine,and amino acid groups,which were generated in their metabolic pathways.After 5 days of PCI,the profiles of serum metabolites were significantly closer between the Danhong-or control-treated groups and that of the healthy group.Furthermore,DHI treatment converted the serum metabolite profile more to that of the healthy group than the control treatment.Conclusion:The beneficial effect of DHI on patients with unstable angina is reflected at the level of serum metabolic biomarkers.
基金This work has been supported by grants from the Taishan Scholars Program(TSQN201812015)the Program for Multidisciplinary Research and Innovation Team of Young Scholars at Shandong University(2020QNQT007).
文摘Objective:The aim of this study is to explore the active ingredients and mechanism of action of danhong injection(DHI)in treating myeloproliferative neoplasms using network pharmacology.Methods:The TCMSP platform and relevant literature were used to search for the active ingredients and targets of Radix Salviae and Carthami Flos in DHI.Disease targets related to myeloproliferative neoplasms were obtained from the GEO database,GeneCards,and DisGeNET database.The queried component targets were normalized using the UniProt database.Potential targets were identified by constructing protein-protein interactions networks using STRING 11.5 and visualized and analyzed using Cytoscape 3.9.1.GO and KEGG analysis were performed using the Metascape platform,and visualization was done using the built-in plug-in CluoGO or SangerBox platforms with Cytoscape 3.9.1.Results:The active ingredients of DHI for treating myeloproliferative neoplasms mainly consist of flavonoids and o-benzoquinones,including quercetin,luteolin,kaempferol,stigmasterol,tanshinone iia,cryptotanshinone,beta-carotene,2-isopropyl-8-methylphenanthrene-3,4-dione,and neocryptotanshinone ii.The potential targets are JUN,TP53,STAT3,AKT1,MAPK1,RELA,TNF,MAPK14,IL6,and FOS.The relevant signaling pathways involved are mainly TNFαsignaling pathway,PI3K-Akt signaling pathway,apoptosis,IL-17 signaling pathway,cellular senescence,MAPK signaling pathway,p53 signaling pathway,JAK-STAT signaling pathway,and NF-kappa B signaling.Conclusions:DHI acts mainly through flavonoids and o-benzoquinones to treat myeloproliferative neoplasms in a multi-targeted and multi-pathway manner.
