AIM:To explore the protective effect and the relevant mechanisms of Fufang Biejia Ruangan Pills(FFBJRGP)on hepatic fibrosis in vivo and in vitro.METHODS:Hepatic fibrosis was induced by carbon tetrachloride composite f...AIM:To explore the protective effect and the relevant mechanisms of Fufang Biejia Ruangan Pills(FFBJRGP)on hepatic fibrosis in vivo and in vitro.METHODS:Hepatic fibrosis was induced by carbon tetrachloride composite factors.Adult Wistar rats were randomly divided into four groups:normal control group;hepatic fibrosis model group;FFBJRGP-treated group at a daily dose of 0.55 g/kg;and colchicinetreated group at a daily dose of 0.1 g/kg.The effects of FFBJRGP on liver function,serum levels of hyaluronic acid(HA),typeⅣcollagen(CⅣ),typeⅢprocollagen(PCⅢ),laminin(LN),histopathology,and expression of transforming growth factor(TGF-β1)and Smad3 in hepatic fibrosis were evaluated in vivo.The effects of FFBJRGP on survival rate,hydroxyproline content and cell cycle distribution were further detected in vitro.RESULTS:Compared with the hepatic fibrosis model group,rats treated with FFBJRGP showed a reduction in hepatic collagen deposition and improvement in hepatic lesions.Compared with those of the model group,the activities of alanine aminotransferase(62.0±23.7 U/L)and aspartate aminotransferase(98.8±40.0 U/L)in the FFBJRGP-treated group were decreased(50.02±3.7 U/L and 57.2±30.0 U/L,respectively,P<0.01).Compared with those in the model group,the levels of PCⅢ(35.73±17.90 g/mL),HA(563.82±335.54 ng/mL),LN(89.57±7.59 ng/mL)and CⅣ(29.20±6.17ng/mL)were decreased to 30.18±9.41,456.18±410.83,85.46±7.51 and 28.02±9.45 ng/mL,respectively.Reverse-transcriptase polymerase chain reaction and Western blotting also revealed that expression of TGF-β1 and Smad3 were down-regulated in vivo.Cell proliferation was inhibited,the level of hydroxyproline was decreased compared with the control group(P<0.01),and the cell cycle was redistributed when exposed to FFBJRGP in vitro.CONCLUSION:FFBJRGP inhibits hepatic fibrosis in vivo and in vitro,which is probably associated with downregulation of fibrogenic signal transduction of the TGF-β-Smad pathway.展开更多
[Objectives]To establish an High Performance Liquid Chromatography(HPLC)method for simultaneous determination of four amino acids in Fufang Ejiao Buxue Granule.[Methods]The quantitative analysis was carried out with a...[Objectives]To establish an High Performance Liquid Chromatography(HPLC)method for simultaneous determination of four amino acids in Fufang Ejiao Buxue Granule.[Methods]The quantitative analysis was carried out with a Waters Sunfire C 18 column(4.6 mm×250 mm,5μm).A binary mobile solvent was used:mobile solvent A was acetonitrile-0.1 mol/L sodium acetate solution(adjusting pH to 6.5 with 36%acetic acid)(7∶93)and mobile solvent B was acetonitrile-H 2O(4∶1).The mobile phase was delivered at a flow rate of 1.0 mL/min with a gradient elution profile(0-13 min,100%A→93%A;13-17.9 min,93%A→88%A;17.9-29.0 min,88%A→85%A;29-39 min,85%A→66%A;39-45 min,66%A→0%A).The column temperature was at 43℃.The detection wavelength was 254 nm.[Results]The injection volume of L-hydroxyproline,glycine,alanine,and L-proline showed a good linear relationship with the chromatographic peak area in the range of 0.012 to 0.117,0.022 to 0.218,0.010 to 0.097,0.016 to 0.160μg,separately.The average recovery rate(n=6)was 96.4%,97.3%,97.1%,and 99.4%,respectively;the relative standard deviations were 1.2%,1.9%,1.7%,and 0.9%,respectively.[Conclusions]This method is simple in operation and good in reproducibility,and provides a reliable method for controlling the quality of Fufang Ejiao Buxue Granules.展开更多
目的:运用网络药理学和分子对接技术探索复方丹参滴丸防治高血压病的潜在分子机制。方法:在中药系统药理学数据库与分析平台(TCMSP)中,以口服生物利用度和类药性为筛选标准,检索出复方丹参滴丸的化学成分,查询对应靶点;利用Genecards数...目的:运用网络药理学和分子对接技术探索复方丹参滴丸防治高血压病的潜在分子机制。方法:在中药系统药理学数据库与分析平台(TCMSP)中,以口服生物利用度和类药性为筛选标准,检索出复方丹参滴丸的化学成分,查询对应靶点;利用Genecards数据库查询高血压病相关靶点,运用Venny数据库得到药物与疾病交集靶点,利用Cytoscape 3.9.1软件构建“药物-活性成分-靶点”网络图及蛋白质-蛋白质相互作用(PPI)关系。通过Metascape和微生信平台对药物与疾病的交集靶点进行京都基因与基因组百科全书(KEGG)通路富集分析及基因本体论(GO)分析,采用Cytoscape 3.9.1软件对筛选的核心成分和核心靶点进行分子对接验证并使用PyMol软件对结果进行可视化分析。结果:共获复方丹参滴丸活性成分76种、相关靶点1062个、复方丹参滴丸与高血压病共有靶点272个,木犀草素、槲皮素、乙酸冰片酯等是复方丹参滴丸防治高血压病的重要化合物,主要通过癌症通路、晚期糖基化终末产物(Advanced Glycation End Product,AGE)-晚期糖基化终末产物受体(Advanced Glycation End Product Receptor,RAGE)信号通路、流体剪切应力与动脉粥样硬化等作用于丝氨酸/苏氨酸蛋白激酶1(Akt Serine/Threonine Kinase 1,AKT1)、Jun原癌基因(Jun Proto-Oncogene,JUN)、丝裂原活化蛋白激酶(Mitogen-Activated Protein Kinase,MAPK)1、MAPK3、信号转导及转录激活蛋白3(Signal Transducer and Activator of Transcription 3,STAT3)等靶点以发挥作用。结论:通过网络药理学证实了复方丹参滴丸多成分、多靶点、多途径的作用规律,为后续临床研究提供了理论依据。展开更多
基金Supported by The Major Project of Applied Basic Research Plan of the Scientific and Technological Department of TianjinChinaNo.06YFJZJC 02900
文摘AIM:To explore the protective effect and the relevant mechanisms of Fufang Biejia Ruangan Pills(FFBJRGP)on hepatic fibrosis in vivo and in vitro.METHODS:Hepatic fibrosis was induced by carbon tetrachloride composite factors.Adult Wistar rats were randomly divided into four groups:normal control group;hepatic fibrosis model group;FFBJRGP-treated group at a daily dose of 0.55 g/kg;and colchicinetreated group at a daily dose of 0.1 g/kg.The effects of FFBJRGP on liver function,serum levels of hyaluronic acid(HA),typeⅣcollagen(CⅣ),typeⅢprocollagen(PCⅢ),laminin(LN),histopathology,and expression of transforming growth factor(TGF-β1)and Smad3 in hepatic fibrosis were evaluated in vivo.The effects of FFBJRGP on survival rate,hydroxyproline content and cell cycle distribution were further detected in vitro.RESULTS:Compared with the hepatic fibrosis model group,rats treated with FFBJRGP showed a reduction in hepatic collagen deposition and improvement in hepatic lesions.Compared with those of the model group,the activities of alanine aminotransferase(62.0±23.7 U/L)and aspartate aminotransferase(98.8±40.0 U/L)in the FFBJRGP-treated group were decreased(50.02±3.7 U/L and 57.2±30.0 U/L,respectively,P<0.01).Compared with those in the model group,the levels of PCⅢ(35.73±17.90 g/mL),HA(563.82±335.54 ng/mL),LN(89.57±7.59 ng/mL)and CⅣ(29.20±6.17ng/mL)were decreased to 30.18±9.41,456.18±410.83,85.46±7.51 and 28.02±9.45 ng/mL,respectively.Reverse-transcriptase polymerase chain reaction and Western blotting also revealed that expression of TGF-β1 and Smad3 were down-regulated in vivo.Cell proliferation was inhibited,the level of hydroxyproline was decreased compared with the control group(P<0.01),and the cell cycle was redistributed when exposed to FFBJRGP in vitro.CONCLUSION:FFBJRGP inhibits hepatic fibrosis in vivo and in vitro,which is probably associated with downregulation of fibrogenic signal transduction of the TGF-β-Smad pathway.
文摘[Objectives]To establish an High Performance Liquid Chromatography(HPLC)method for simultaneous determination of four amino acids in Fufang Ejiao Buxue Granule.[Methods]The quantitative analysis was carried out with a Waters Sunfire C 18 column(4.6 mm×250 mm,5μm).A binary mobile solvent was used:mobile solvent A was acetonitrile-0.1 mol/L sodium acetate solution(adjusting pH to 6.5 with 36%acetic acid)(7∶93)and mobile solvent B was acetonitrile-H 2O(4∶1).The mobile phase was delivered at a flow rate of 1.0 mL/min with a gradient elution profile(0-13 min,100%A→93%A;13-17.9 min,93%A→88%A;17.9-29.0 min,88%A→85%A;29-39 min,85%A→66%A;39-45 min,66%A→0%A).The column temperature was at 43℃.The detection wavelength was 254 nm.[Results]The injection volume of L-hydroxyproline,glycine,alanine,and L-proline showed a good linear relationship with the chromatographic peak area in the range of 0.012 to 0.117,0.022 to 0.218,0.010 to 0.097,0.016 to 0.160μg,separately.The average recovery rate(n=6)was 96.4%,97.3%,97.1%,and 99.4%,respectively;the relative standard deviations were 1.2%,1.9%,1.7%,and 0.9%,respectively.[Conclusions]This method is simple in operation and good in reproducibility,and provides a reliable method for controlling the quality of Fufang Ejiao Buxue Granules.
文摘目的:运用网络药理学和分子对接技术探索复方丹参滴丸防治高血压病的潜在分子机制。方法:在中药系统药理学数据库与分析平台(TCMSP)中,以口服生物利用度和类药性为筛选标准,检索出复方丹参滴丸的化学成分,查询对应靶点;利用Genecards数据库查询高血压病相关靶点,运用Venny数据库得到药物与疾病交集靶点,利用Cytoscape 3.9.1软件构建“药物-活性成分-靶点”网络图及蛋白质-蛋白质相互作用(PPI)关系。通过Metascape和微生信平台对药物与疾病的交集靶点进行京都基因与基因组百科全书(KEGG)通路富集分析及基因本体论(GO)分析,采用Cytoscape 3.9.1软件对筛选的核心成分和核心靶点进行分子对接验证并使用PyMol软件对结果进行可视化分析。结果:共获复方丹参滴丸活性成分76种、相关靶点1062个、复方丹参滴丸与高血压病共有靶点272个,木犀草素、槲皮素、乙酸冰片酯等是复方丹参滴丸防治高血压病的重要化合物,主要通过癌症通路、晚期糖基化终末产物(Advanced Glycation End Product,AGE)-晚期糖基化终末产物受体(Advanced Glycation End Product Receptor,RAGE)信号通路、流体剪切应力与动脉粥样硬化等作用于丝氨酸/苏氨酸蛋白激酶1(Akt Serine/Threonine Kinase 1,AKT1)、Jun原癌基因(Jun Proto-Oncogene,JUN)、丝裂原活化蛋白激酶(Mitogen-Activated Protein Kinase,MAPK)1、MAPK3、信号转导及转录激活蛋白3(Signal Transducer and Activator of Transcription 3,STAT3)等靶点以发挥作用。结论:通过网络药理学证实了复方丹参滴丸多成分、多靶点、多途径的作用规律,为后续临床研究提供了理论依据。
