OBJECTIVE:To analyze the effect and molecular mechanism of Gehua Jiejiu Dizhi decoction(葛花解酒涤脂汤,GJDD)on alcoholic fatty live disease(AFLD)by using proteomic methods.METHODS:The male C57BL/6J mouse were randomly...OBJECTIVE:To analyze the effect and molecular mechanism of Gehua Jiejiu Dizhi decoction(葛花解酒涤脂汤,GJDD)on alcoholic fatty live disease(AFLD)by using proteomic methods.METHODS:The male C57BL/6J mouse were randomly divided into four groups:control group,model group,GJDD group and resveratrol group.After the AFLD model was successfully prepared by intragastric administration of alcohol once on the basis of the Lieber-DeCarli classical method,the GJDD group and resveratrol group were intragastrically administered with GJDD(4900 mg/kg)and resveratrol(400 mg/kg)respectively,once a day for 9 d.The fat deposition of liver tissue was observed and evaluated by oil red O(ORO)staining.4DLabel-free quantitative proteome method was used to determine and quantify the protein expression in liver tissue of each experimental group.The differentially expressed proteins were screened according to protein expression differential multiples,and then analyzed by Gene ontology classification and Kyoto Encyclopedia of Genes and Genomes pathway enrichment.Finally,expression validation of the differentially co-expressed proteins from control group,model group and GJDD group were verified by targeted proteomics quantification techniques.RESULTS:In semiquantitative analyses of ORO,all kinds of steatosis(ToS,MaS,and MiS)were evaluated higher in AFLD mice compared to those in GJDD or resveratroltreated mice.4DLabel-free proteomics analysis results showed that a total of 4513 proteins were identified,of which 3763 proteins were quantified and 946 differentially expressed proteins were screened.Compared with the control group,145 proteins were up-regulated and 148 proteins were down-regulated in the liver tissue of model group.In addition,compared with the model group,92 proteins were up-regulated and 135 proteins were downregulated in the liver tissue of the GJDD group.15 differentially co-expressed proteins were found between every two groups(model group vs control group,GJDD group vs model group and GJDD group vs control group),which were involved in many biological processes.Among them,11 differentially co-expressed key proteins(Aox3,H1-5,Fabp5,Ces3a,Nudt7,Serpinb1a,Fkbp11,Rpl22l1,Keg1,Acss2 and Slco1a1)were further identified by targeted proteomic quantitative technology and their expression patterns were consistent with the results of 4D label-free proteomic analysis.CONCLUSIONS:Our study provided proteomics-based evidence that GJDD alleviated AFLD by modulating liver protein expression,likely through the modulation of lipid metabolism,bile acid metabolism and with exertion of antioxidant stress.展开更多
目的:通过网络药理学、生物信息学、分子对接及实验方法探讨葛花解酲方通过调控铁死亡抑制上皮间质转化(EMT)干预肝细胞癌(HCC)的机制。方法:通过中药系统药理学数据库与分析平台(TCMSP)、蛋白质数据库(Uniprot)、基因表达数据库(GEO)...目的:通过网络药理学、生物信息学、分子对接及实验方法探讨葛花解酲方通过调控铁死亡抑制上皮间质转化(EMT)干预肝细胞癌(HCC)的机制。方法:通过中药系统药理学数据库与分析平台(TCMSP)、蛋白质数据库(Uniprot)、基因表达数据库(GEO)、蛋白质相互作用网络数据库(String)等筛选出药物疾病靶点;通过生物信息学分析差异基因表达、人类蛋白质图谱(THPA)数据库蛋白表达、肝癌临床相关性、生存预后、基因集富(GSEA)筛选关键靶点;通过Discovery Studio 2016分子对接软件筛选关键靶点对应的化合物;通过Transwell和Western blot法探讨葛花解酲方含药血清对HepG2细胞的EMT作用。结果:获得葛花解酲方-铁死亡-肝癌交集靶点13个;通过差异基因表达分析、THPA数据库蛋白表达分析、肝癌临床相关性分析、生存预后分析最终筛选得出1个关键靶点为雄激素受体(AR)。GSEA富集分析发现AR与上皮间质转化(EMT)的细胞黏附过程密切相关。分子对接结果显示,AR与脱氧三尖杉酯碱、3-甲氧基葛根素、葛根素和染料木素8-C-葡萄糖苷成分有较高的亲合力。与空白对照组比较,葛花解酲方含药血清可显著降低HepG2细胞的迁移率,显著上调HepG2细胞中E-cadherin(上皮标志物)、AR的表达(P<0.01),下调N-cadherin(间质标志物)、谷胱甘肽过氧化酶4(GPX4,铁死亡标志物)的表达(P<0.01)。结论:葛花解酲方可以通过多种有效成分作用于AR调控铁死亡抑制EMT而干预HCC。展开更多
目的:探讨葛花解酲汤对小鼠醉酒模型的解酒护肝作用。方法:将小鼠随机分为对照组、醉酒模型组、葛花解酲汤组,各组小鼠分别按5 m L/kg和15 m L/kg体质量灌胃56%红星二锅头建立小鼠醉酒模型,葛花解酲汤组以葛花解酲汤灌胃,对照组以蒸馏...目的:探讨葛花解酲汤对小鼠醉酒模型的解酒护肝作用。方法:将小鼠随机分为对照组、醉酒模型组、葛花解酲汤组,各组小鼠分别按5 m L/kg和15 m L/kg体质量灌胃56%红星二锅头建立小鼠醉酒模型,葛花解酲汤组以葛花解酲汤灌胃,对照组以蒸馏水灌胃,以自主活动次数、翻正反应耐受时间与持续时间、醉酒睡眠潜伏时间与持续时间、肝脏系数和丙氨酸氨基转移酶(ALT)含量作为考察指标。结果:醉酒初期葛花解酲汤组小鼠自主活动次数明显高于对照组而低于醉酒模型组;大量醉酒时葛花解酲汤组小鼠较醉酒模型组小鼠翻正反应耐受时间和睡眠潜伏时间明显延长;葛花解酲汤组和对照组小鼠的肝脏系数明显低于醉酒模型组;葛花解酲汤组小鼠ALT含量明显低于醉酒模型组而高于对照组。结论:葛花解酲汤具有显著的醒酒、防醉与保肝护肝作用。展开更多
基金National Science Foundation-funded Project:the Study on the Changes of Energy Metabolism and Molecular Regulation Mechanism of Alcoholic Fatty Liver based on Sirtuins1-Adenosine Monophosphate-Activated Protein Kinase Signal System and the Intervention of Gehua Jiejiu dizhi decoction(No.81660752)Basic Research Project of Guizhou Provincial Science and Technology Plan:Study on the Mechanism of Sirtuins1 Mediated Deacetylation in the Regulation of Alcoholic Fatty Liver Metabolism and the Intervention of Gehua Jiejiu Dizhi Tang[QianKeHe Fundamentals-ZK[2023]General 410]。
文摘OBJECTIVE:To analyze the effect and molecular mechanism of Gehua Jiejiu Dizhi decoction(葛花解酒涤脂汤,GJDD)on alcoholic fatty live disease(AFLD)by using proteomic methods.METHODS:The male C57BL/6J mouse were randomly divided into four groups:control group,model group,GJDD group and resveratrol group.After the AFLD model was successfully prepared by intragastric administration of alcohol once on the basis of the Lieber-DeCarli classical method,the GJDD group and resveratrol group were intragastrically administered with GJDD(4900 mg/kg)and resveratrol(400 mg/kg)respectively,once a day for 9 d.The fat deposition of liver tissue was observed and evaluated by oil red O(ORO)staining.4DLabel-free quantitative proteome method was used to determine and quantify the protein expression in liver tissue of each experimental group.The differentially expressed proteins were screened according to protein expression differential multiples,and then analyzed by Gene ontology classification and Kyoto Encyclopedia of Genes and Genomes pathway enrichment.Finally,expression validation of the differentially co-expressed proteins from control group,model group and GJDD group were verified by targeted proteomics quantification techniques.RESULTS:In semiquantitative analyses of ORO,all kinds of steatosis(ToS,MaS,and MiS)were evaluated higher in AFLD mice compared to those in GJDD or resveratroltreated mice.4DLabel-free proteomics analysis results showed that a total of 4513 proteins were identified,of which 3763 proteins were quantified and 946 differentially expressed proteins were screened.Compared with the control group,145 proteins were up-regulated and 148 proteins were down-regulated in the liver tissue of model group.In addition,compared with the model group,92 proteins were up-regulated and 135 proteins were downregulated in the liver tissue of the GJDD group.15 differentially co-expressed proteins were found between every two groups(model group vs control group,GJDD group vs model group and GJDD group vs control group),which were involved in many biological processes.Among them,11 differentially co-expressed key proteins(Aox3,H1-5,Fabp5,Ces3a,Nudt7,Serpinb1a,Fkbp11,Rpl22l1,Keg1,Acss2 and Slco1a1)were further identified by targeted proteomic quantitative technology and their expression patterns were consistent with the results of 4D label-free proteomic analysis.CONCLUSIONS:Our study provided proteomics-based evidence that GJDD alleviated AFLD by modulating liver protein expression,likely through the modulation of lipid metabolism,bile acid metabolism and with exertion of antioxidant stress.
文摘目的:通过网络药理学、生物信息学、分子对接及实验方法探讨葛花解酲方通过调控铁死亡抑制上皮间质转化(EMT)干预肝细胞癌(HCC)的机制。方法:通过中药系统药理学数据库与分析平台(TCMSP)、蛋白质数据库(Uniprot)、基因表达数据库(GEO)、蛋白质相互作用网络数据库(String)等筛选出药物疾病靶点;通过生物信息学分析差异基因表达、人类蛋白质图谱(THPA)数据库蛋白表达、肝癌临床相关性、生存预后、基因集富(GSEA)筛选关键靶点;通过Discovery Studio 2016分子对接软件筛选关键靶点对应的化合物;通过Transwell和Western blot法探讨葛花解酲方含药血清对HepG2细胞的EMT作用。结果:获得葛花解酲方-铁死亡-肝癌交集靶点13个;通过差异基因表达分析、THPA数据库蛋白表达分析、肝癌临床相关性分析、生存预后分析最终筛选得出1个关键靶点为雄激素受体(AR)。GSEA富集分析发现AR与上皮间质转化(EMT)的细胞黏附过程密切相关。分子对接结果显示,AR与脱氧三尖杉酯碱、3-甲氧基葛根素、葛根素和染料木素8-C-葡萄糖苷成分有较高的亲合力。与空白对照组比较,葛花解酲方含药血清可显著降低HepG2细胞的迁移率,显著上调HepG2细胞中E-cadherin(上皮标志物)、AR的表达(P<0.01),下调N-cadherin(间质标志物)、谷胱甘肽过氧化酶4(GPX4,铁死亡标志物)的表达(P<0.01)。结论:葛花解酲方可以通过多种有效成分作用于AR调控铁死亡抑制EMT而干预HCC。
文摘目的:探讨葛花解酲汤对小鼠醉酒模型的解酒护肝作用。方法:将小鼠随机分为对照组、醉酒模型组、葛花解酲汤组,各组小鼠分别按5 m L/kg和15 m L/kg体质量灌胃56%红星二锅头建立小鼠醉酒模型,葛花解酲汤组以葛花解酲汤灌胃,对照组以蒸馏水灌胃,以自主活动次数、翻正反应耐受时间与持续时间、醉酒睡眠潜伏时间与持续时间、肝脏系数和丙氨酸氨基转移酶(ALT)含量作为考察指标。结果:醉酒初期葛花解酲汤组小鼠自主活动次数明显高于对照组而低于醉酒模型组;大量醉酒时葛花解酲汤组小鼠较醉酒模型组小鼠翻正反应耐受时间和睡眠潜伏时间明显延长;葛花解酲汤组和对照组小鼠的肝脏系数明显低于醉酒模型组;葛花解酲汤组小鼠ALT含量明显低于醉酒模型组而高于对照组。结论:葛花解酲汤具有显著的醒酒、防醉与保肝护肝作用。
