[Objectives]This study was conducted to establish the method for simultaneous determination of six active components.[Methods]Simultaneous determination of chlorogenic acid,phellodendrine,magnoflorine,jatrorrhizine,pa...[Objectives]This study was conducted to establish the method for simultaneous determination of six active components.[Methods]Simultaneous determination of chlorogenic acid,phellodendrine,magnoflorine,jatrorrhizine,palmatine and berberine in Cortex Phellodendri was carried out by HPLC with a Diamonsil C18(4.6 mm×250 mm,5μm)column was used.The mobile phase was acetonitrile-water(1‰acetic acid,2 mmol ammonium acetate)solution in gradient elution.The detection wavelength was set at 280 nm,and the column temperature was kept at 25℃and the flow rate was 1 ml/min.[Results]The linear ranges of chlorogenic acid,phellodendrine,magnoflorine,jatrorrhizine,palmatine and berberine were 20.00-320.00,18.75-130.00,25.00-200.00,5.00-100.00,20.00-200.00,and 0.09-1.80 mg/L,respectively.The average recovery was 98.1%,99.4%,97.5%,97.3%,104.0%,and 98.5%,respectively;and the RSDs were 0.5%,0.6%,0.8%,1.0%,1.4%,and 0.9%,respectively.[Conclusions]The method is convenient,stable,reliable and suitable for quality control of Cortex Phellodendri.展开更多
A combined method of high performance liquid chromatograph-elecrtrospray-ionization mass spectrometer（HPLC-ESI-MS/MS） coupled with a photodiode array detector（HPLC-DAD） and principal component analysis（PCA） was ...A combined method of high performance liquid chromatograph-elecrtrospray-ionization mass spectrometer（HPLC-ESI-MS/MS） coupled with a photodiode array detector（HPLC-DAD） and principal component analysis（PCA） was applied to the qualitative and quantitative analyses of alkaloids in Cortex Phellodendri（CP） samples, and to the differentiation of two species of CP, Cortex Phellodendri Chinensis（CPC） and Cortex Phellodendri Amurensis（CPA）. Twenty-two peaks appeared in the HPLC-MS base peak chromatogram of CP detected by the HPLC-ESI-MS/MS analysis, and the alkaloids were identified according to the MSn data, the known MS fragmentation rules and the literature data. Five alkaloids including berberine, palmatine, jatrorrhizine, phellodendrine and magnoflorine were simultaneously determinated by the HPLC-DAD. Berberine was the primary component in all CP samples, and the contents of berberine and palmatine were exploited to be two critical parameters for effective discrimination between the two species of CP. The average content of berberine in CPC（58.75 mg/g） was higher than that in CPA（9.16 mg/g）, while the content of palmatine was less, only 0.25 mg/g in CPC and 4.19 mg/g in CPA. With the use of PCA, samples datasets were separated successfully into two different clusters corresponding to the two species, and berberine, pahnatine, phellodendrine and magnoflorine contribute most to the above mentioned calssifying . The proposed method oroved to be a useful tool in the aualitv control of Chinese herbal medicines.展开更多
Coptidis Rhizoma(Chinese:Huanglian)and Phellodendri Chinensis Cortex(Chinese:Huangbo)are widely used Traditional Chinese Medicine,and often used in combination because of their similar pharmacological effects in clini...Coptidis Rhizoma(Chinese:Huanglian)and Phellodendri Chinensis Cortex(Chinese:Huangbo)are widely used Traditional Chinese Medicine,and often used in combination because of their similar pharmacological effects in clinical practice.However,the quality control methods of the two drugs are different and complicated,which is time consuming and laborious in practical application.In this paper,rapid and simultaneous determination of moisture and berberine in Coptidis Rhizoma(CR)and Phellodendri Chinensis Cortex(PC)was realized by near-infrared spectroscopy(NIRs)combined with global models.Competitive adaptive reweighted sampling(CARS)and successive projection algorithm(SPA)method were applied for variable selection.Principal component analysis(PCA)and partial least squares regression method(PLSR)were applied for qualitative and quantitative analysis,respectively.The characteristic variables of berberine showed similarity and consistency in distribution,providing basis for the global models.For moisture content,the global model had relative standard error of prediction set(RSEP)value of 3.04%and 2.53%for CR and PC,respectively.For berberine content,the global model had RSEP value of 5.41%and 3.97%for CR and PC,respectively.These results indicated the global models based on CARS-PLS method achieved satisfactory prediction for moisture and berberine content,improving the determination e±ciency.Furthermore,the greater range and larger number of samples enhanced the reliance of the global model.The NIRs combined with global models could be a powerful tool for quality control of CR and PC.展开更多
Traditional Chinese medicine(TCM)Phellodendri Chinensis Cortex(PCC)has been used for the treatment of human hepatocellular carcinoma,but the underlying mechanisms are still unclear.In this study,15 active compositions...Traditional Chinese medicine(TCM)Phellodendri Chinensis Cortex(PCC)has been used for the treatment of human hepatocellular carcinoma,but the underlying mechanisms are still unclear.In this study,15 active compositions of PCC were obtained from Traditional Chinese Medicine Systems Pharmacology Database(TCMSP),and 505 putative identified targets of PCC were screened by Swiss Target Prediction server.Next,HCC data was downloaded from Drugbank and GeneCards databases.Furtherly,45 common targets were revealed.The network diagrams of the active component-target network,protein-protein interaction(PPI)network and active component-target-pathway network were constructed using Cytoscape software.The analysis of the network results showed that the active ingredients of PCC,such as berberine,obacunone,rutaecarpine,candletoxin A,palmatine,isocorypalmine,quercetin,and(S)-Canadine,had a good binding activity with more targets.Additionally,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses revealed that common targets were significantly enriched in Ras signaling pathway,ErbB signaling pathway,and Mammalian target of rapamycin(mTOR)signaling pathway.Altogether,the multi-component,multi-target,and multi-pathway characteristics of PCC provided a reference for the in-depth study of the mechanism of PCC in the treatment of HCC.展开更多
目的:基于网络药理学和分子对接方法,确定复方黄柏液治疗的Ⅲ度烧伤肉芽组织愈合的有效活性成分、关键靶点和潜在的药理学机制,并进行肉芽组织成纤维细胞的初步验证。方法:从公共数据库中药系统药理学分析平台(TCMSP)检索复方黄柏液组...目的:基于网络药理学和分子对接方法,确定复方黄柏液治疗的Ⅲ度烧伤肉芽组织愈合的有效活性成分、关键靶点和潜在的药理学机制,并进行肉芽组织成纤维细胞的初步验证。方法:从公共数据库中药系统药理学分析平台(TCMSP)检索复方黄柏液组成成分连翘、黄柏、金银花的有效成分和靶点;GeneCards、OMIM数据库检索“Ⅲ度烧伤”疾病相关靶点。通过生物信息学分析,包括蛋白质-蛋白质相互作用(Protein-proteininteraction,PPI)以及基因本体(Gene ontology,GO)和京都基因和基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析,获得了关键的有效成分、核心靶点和相关信号通路;DiscoveryStudio分子对接分析有效成分化合物与靶蛋白的结合。0.5%的DMSO溶液处理的成纤维细胞记为对照组;槲皮素(40μmol/ml)处理的成纤维细胞记为槲皮素组。采用CCK8法、Transwell实验检测细胞增殖、迁移侵袭;WB试验检测细胞p-PI3K、p-Akt蛋白。结果:共筛选出74个有效成分,331个作用靶点,AKT1为潜在的治疗靶点,木犀草素、山柰酚、槲皮素、汉黄芩素、丹皮酚为潜在的候选药物。PI3K-AKT信号通路可能在复方黄柏液治疗Ⅲ度烧伤中发挥关键作用;分子对接表明槲皮素与AKT1结合最好。与对照组相比,槲皮素组成纤维细胞增殖、迁移侵袭均显著降低,p-PI3K、p-Akt蛋白表达也显著降低(P<0.05)。结论:复方黄柏液促进Ⅲ度烧伤患者肉芽组织形成的生物活性成分为槲皮素,潜在通路为PI3K-AKT信号通路,为复方黄柏液治疗Ⅲ度烧伤的研究提供了思路。展开更多
目的:利用网络药理学方法和分子对接技术,分析“黄连-黄芩-黄柏”(简称“三黄”)治疗细菌性痢疾的潜在活性成分和作用机制,为后续的新药开发研究提供依据。方法:利用中药系统药理学平台(Traditional Chinese Medicine Systems Pharmacol...目的:利用网络药理学方法和分子对接技术,分析“黄连-黄芩-黄柏”(简称“三黄”)治疗细菌性痢疾的潜在活性成分和作用机制,为后续的新药开发研究提供依据。方法:利用中药系统药理学平台(Traditional Chinese Medicine Systems Pharmacology Database,TCMSP)获取“三黄”的活性化合物及其潜在作用靶点,再从GeneCards、CTD、TTD和drugbank等数据库获取细菌性痢疾的疾病靶点,然后确定“三黄”治疗细菌性痢疾的潜在作用靶点,随后进行蛋白质-蛋白质相互作用(protein-protein interaction,PPI)分析,以筛选出核心靶点,进而通过GO功能分析和KEGG通路富集分析筛选“三黄”治疗细菌性痢疾的生物过程和相关通路,最后通过分子对接处理分析受体与配体间的亲和力。结果:借助TCMSP,共从“三黄”中筛选出87个关键活性成分,而其对应的作用靶点有228个,而在相关数据库中获取到细菌性痢疾的疾病靶点共313个,通过比较分析得到交集靶点34个;通过PPI分析,又从34个交集靶点中筛选得到14个核心靶点;通过KEGG通路富集分析发现,有14条治疗细菌性痢疾的重要信号通路;利用Cytoscape软件,根据34个交集靶点和14条重要信号通路,筛选得到度值最大的前5个活性成分依次为槲皮素、汉黄芩素、黄芩素、β-谷甾醇、异延胡索单酚碱;而分子对接结果显示,肿瘤坏死因子(TNF)、白介素-6(IL-6)和环氧合酶2(PTGS2)与槲皮素、汉黄芩素、黄芩素、β-谷甾醇、异延胡索单酚碱具有较好的亲和力(结合能<-5 kJ/mol)。结论:槲皮素、汉黄芩素、黄芩素、β-谷甾醇、异延胡索单酚碱可能是“三黄”治疗细菌性痢疾的主要潜在活性成分,而TNF、IL-6和PTGS2可能是其主要作用靶点。展开更多
文摘[Objectives]This study was conducted to establish the method for simultaneous determination of six active components.[Methods]Simultaneous determination of chlorogenic acid,phellodendrine,magnoflorine,jatrorrhizine,palmatine and berberine in Cortex Phellodendri was carried out by HPLC with a Diamonsil C18(4.6 mm×250 mm,5μm)column was used.The mobile phase was acetonitrile-water(1‰acetic acid,2 mmol ammonium acetate)solution in gradient elution.The detection wavelength was set at 280 nm,and the column temperature was kept at 25℃and the flow rate was 1 ml/min.[Results]The linear ranges of chlorogenic acid,phellodendrine,magnoflorine,jatrorrhizine,palmatine and berberine were 20.00-320.00,18.75-130.00,25.00-200.00,5.00-100.00,20.00-200.00,and 0.09-1.80 mg/L,respectively.The average recovery was 98.1%,99.4%,97.5%,97.3%,104.0%,and 98.5%,respectively;and the RSDs were 0.5%,0.6%,0.8%,1.0%,1.4%,and 0.9%,respectively.[Conclusions]The method is convenient,stable,reliable and suitable for quality control of Cortex Phellodendri.
基金Supported by the National Natural Science Foundation of China(No30725045)the Foundation of Eleventh Five-Year-Plan of China(No2008ZX09202-002)+1 种基金the Shanghai Leading Academic Discipline Project, China(NoB906)the Scientific Foundation of Shanghai City, China(No07DZ19702)
文摘A combined method of high performance liquid chromatograph-elecrtrospray-ionization mass spectrometer（HPLC-ESI-MS/MS） coupled with a photodiode array detector（HPLC-DAD） and principal component analysis（PCA） was applied to the qualitative and quantitative analyses of alkaloids in Cortex Phellodendri（CP） samples, and to the differentiation of two species of CP, Cortex Phellodendri Chinensis（CPC） and Cortex Phellodendri Amurensis（CPA）. Twenty-two peaks appeared in the HPLC-MS base peak chromatogram of CP detected by the HPLC-ESI-MS/MS analysis, and the alkaloids were identified according to the MSn data, the known MS fragmentation rules and the literature data. Five alkaloids including berberine, palmatine, jatrorrhizine, phellodendrine and magnoflorine were simultaneously determinated by the HPLC-DAD. Berberine was the primary component in all CP samples, and the contents of berberine and palmatine were exploited to be two critical parameters for effective discrimination between the two species of CP. The average content of berberine in CPC（58.75 mg/g） was higher than that in CPA（9.16 mg/g）, while the content of palmatine was less, only 0.25 mg/g in CPC and 4.19 mg/g in CPA. With the use of PCA, samples datasets were separated successfully into two different clusters corresponding to the two species, and berberine, pahnatine, phellodendrine and magnoflorine contribute most to the above mentioned calssifying . The proposed method oroved to be a useful tool in the aualitv control of Chinese herbal medicines.
基金supported by National Major Scientic and Technological Special Project for"Signicant New Drugs Development"(2018ZX09201010).
文摘Coptidis Rhizoma(Chinese:Huanglian)and Phellodendri Chinensis Cortex(Chinese:Huangbo)are widely used Traditional Chinese Medicine,and often used in combination because of their similar pharmacological effects in clinical practice.However,the quality control methods of the two drugs are different and complicated,which is time consuming and laborious in practical application.In this paper,rapid and simultaneous determination of moisture and berberine in Coptidis Rhizoma(CR)and Phellodendri Chinensis Cortex(PC)was realized by near-infrared spectroscopy(NIRs)combined with global models.Competitive adaptive reweighted sampling(CARS)and successive projection algorithm(SPA)method were applied for variable selection.Principal component analysis(PCA)and partial least squares regression method(PLSR)were applied for qualitative and quantitative analysis,respectively.The characteristic variables of berberine showed similarity and consistency in distribution,providing basis for the global models.For moisture content,the global model had relative standard error of prediction set(RSEP)value of 3.04%and 2.53%for CR and PC,respectively.For berberine content,the global model had RSEP value of 5.41%and 3.97%for CR and PC,respectively.These results indicated the global models based on CARS-PLS method achieved satisfactory prediction for moisture and berberine content,improving the determination e±ciency.Furthermore,the greater range and larger number of samples enhanced the reliance of the global model.The NIRs combined with global models could be a powerful tool for quality control of CR and PC.
文摘Traditional Chinese medicine(TCM)Phellodendri Chinensis Cortex(PCC)has been used for the treatment of human hepatocellular carcinoma,but the underlying mechanisms are still unclear.In this study,15 active compositions of PCC were obtained from Traditional Chinese Medicine Systems Pharmacology Database(TCMSP),and 505 putative identified targets of PCC were screened by Swiss Target Prediction server.Next,HCC data was downloaded from Drugbank and GeneCards databases.Furtherly,45 common targets were revealed.The network diagrams of the active component-target network,protein-protein interaction(PPI)network and active component-target-pathway network were constructed using Cytoscape software.The analysis of the network results showed that the active ingredients of PCC,such as berberine,obacunone,rutaecarpine,candletoxin A,palmatine,isocorypalmine,quercetin,and(S)-Canadine,had a good binding activity with more targets.Additionally,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses revealed that common targets were significantly enriched in Ras signaling pathway,ErbB signaling pathway,and Mammalian target of rapamycin(mTOR)signaling pathway.Altogether,the multi-component,multi-target,and multi-pathway characteristics of PCC provided a reference for the in-depth study of the mechanism of PCC in the treatment of HCC.
文摘目的:基于网络药理学和分子对接方法,确定复方黄柏液治疗的Ⅲ度烧伤肉芽组织愈合的有效活性成分、关键靶点和潜在的药理学机制,并进行肉芽组织成纤维细胞的初步验证。方法:从公共数据库中药系统药理学分析平台(TCMSP)检索复方黄柏液组成成分连翘、黄柏、金银花的有效成分和靶点;GeneCards、OMIM数据库检索“Ⅲ度烧伤”疾病相关靶点。通过生物信息学分析,包括蛋白质-蛋白质相互作用(Protein-proteininteraction,PPI)以及基因本体(Gene ontology,GO)和京都基因和基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析,获得了关键的有效成分、核心靶点和相关信号通路;DiscoveryStudio分子对接分析有效成分化合物与靶蛋白的结合。0.5%的DMSO溶液处理的成纤维细胞记为对照组;槲皮素(40μmol/ml)处理的成纤维细胞记为槲皮素组。采用CCK8法、Transwell实验检测细胞增殖、迁移侵袭;WB试验检测细胞p-PI3K、p-Akt蛋白。结果:共筛选出74个有效成分,331个作用靶点,AKT1为潜在的治疗靶点,木犀草素、山柰酚、槲皮素、汉黄芩素、丹皮酚为潜在的候选药物。PI3K-AKT信号通路可能在复方黄柏液治疗Ⅲ度烧伤中发挥关键作用;分子对接表明槲皮素与AKT1结合最好。与对照组相比,槲皮素组成纤维细胞增殖、迁移侵袭均显著降低,p-PI3K、p-Akt蛋白表达也显著降低(P<0.05)。结论:复方黄柏液促进Ⅲ度烧伤患者肉芽组织形成的生物活性成分为槲皮素,潜在通路为PI3K-AKT信号通路,为复方黄柏液治疗Ⅲ度烧伤的研究提供了思路。
文摘目的:利用网络药理学方法和分子对接技术,分析“黄连-黄芩-黄柏”(简称“三黄”)治疗细菌性痢疾的潜在活性成分和作用机制,为后续的新药开发研究提供依据。方法:利用中药系统药理学平台(Traditional Chinese Medicine Systems Pharmacology Database,TCMSP)获取“三黄”的活性化合物及其潜在作用靶点,再从GeneCards、CTD、TTD和drugbank等数据库获取细菌性痢疾的疾病靶点,然后确定“三黄”治疗细菌性痢疾的潜在作用靶点,随后进行蛋白质-蛋白质相互作用(protein-protein interaction,PPI)分析,以筛选出核心靶点,进而通过GO功能分析和KEGG通路富集分析筛选“三黄”治疗细菌性痢疾的生物过程和相关通路,最后通过分子对接处理分析受体与配体间的亲和力。结果:借助TCMSP,共从“三黄”中筛选出87个关键活性成分,而其对应的作用靶点有228个,而在相关数据库中获取到细菌性痢疾的疾病靶点共313个,通过比较分析得到交集靶点34个;通过PPI分析,又从34个交集靶点中筛选得到14个核心靶点;通过KEGG通路富集分析发现,有14条治疗细菌性痢疾的重要信号通路;利用Cytoscape软件,根据34个交集靶点和14条重要信号通路,筛选得到度值最大的前5个活性成分依次为槲皮素、汉黄芩素、黄芩素、β-谷甾醇、异延胡索单酚碱;而分子对接结果显示,肿瘤坏死因子(TNF)、白介素-6(IL-6)和环氧合酶2(PTGS2)与槲皮素、汉黄芩素、黄芩素、β-谷甾醇、异延胡索单酚碱具有较好的亲和力(结合能<-5 kJ/mol)。结论:槲皮素、汉黄芩素、黄芩素、β-谷甾醇、异延胡索单酚碱可能是“三黄”治疗细菌性痢疾的主要潜在活性成分,而TNF、IL-6和PTGS2可能是其主要作用靶点。
