目的建立基于高分辨质谱、全球天然产物分子网络(global natural products social molecular networking,GNPS)和网络药理学的中药治疗共病物质基础研究策略,并对开心散治疗抑郁症-阿尔茨海默病的物质基础进行系统分析。方法采用超高效...目的建立基于高分辨质谱、全球天然产物分子网络(global natural products social molecular networking,GNPS)和网络药理学的中药治疗共病物质基础研究策略,并对开心散治疗抑郁症-阿尔茨海默病的物质基础进行系统分析。方法采用超高效液相色谱-轨道阱串联质谱(ultra-performance liquid chromatography-orbitrap tandem mass spectrometry,UPLCOrbitrap-MS/MS)对开心散提取物及其血浆和脑组织暴露样品进行非靶向二级质谱分析。然后,利用GNPS平台,基于开心散提取物和血浆样品所得质谱数据构建分子网络,结合化合物的精确质量数、保留时间、碎片信息、质谱裂解规律,快速鉴定开心散血浆暴露成分。基于血浆暴露成分,结合脑组织样品的高分辨质谱数据,鉴定开心散入脑成分。结合脑组织暴露成分和网络药理学技术探究开心散治疗抑郁症-阿尔茨海默病共病物质基础。结果从血浆中鉴定出开心散暴露成分共34个,从脑组织中鉴定出开心散暴露成分共21个,并筛选出开心散中具有治疗抑郁症-阿尔茨海默病共病活性的成分共19个,涉及皂苷类成分9个、寡糖酯类成分9个、酮类成分1个。结论建立了中药共病物质基础快速发现的新策略,为中药“异病同治”研究提供方法支撑,且为开心散治疗抑郁症-阿尔茨海默病共病的物质基础阐释及其临床应用提供数据支撑。展开更多
Background:Wenqing Yin(WQY)is a classic prescription used to treat skin diseases like atopic dermatitis(AD)in China,and the aim of this study is to investigate the therapeutic effects and molecular mechanisms of WQY o...Background:Wenqing Yin(WQY)is a classic prescription used to treat skin diseases like atopic dermatitis(AD)in China,and the aim of this study is to investigate the therapeutic effects and molecular mechanisms of WQY on AD.Methods:The DNFB-induced mouse models of AD were established to investigate the therapeutic effects of WQY on AD.The symptoms of AD in the ears and backs of the mice were assessed,while inflammatory factors in the ear were quantified using quantitative real-time-polymerase chain reaction(qRT-PCR),and the percentages of CD4^(+)and CD8^(+)cells in the spleen were analyzed through flow cytometry.The compounds in WQY were identified using ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)analysis and the key targets and pathways of WQY to treat AD were predicted by network pharmacology.Subsequently,the key genes were tested and verified by qRT-PCR,and the potential active components and target proteins were verified by molecular docking.Results:WQY relieved the AD symptoms and histopathological injuries in the ear and back skin of mice with AD.Meanwhile,WQY significantly reduced the levels of inflammatory factors IL-6 and IL-1βin ear tissue,as well as the ratio of CD4^(+)/CD8^(+)cells in spleen.Additionally,a total of 142 compounds were identified from the water extract of WQY by UPLC-Orbitrap-MS/MS.39 key targets related to AD were screened out by network pharmacology methods.The KEGG analysis indicated that the effects of WQY were primarily mediated through pathways associated with Toll-like receptor signaling and T cell receptor signaling.Moreover,the results of qRT-PCR demonstrated that WQY significantly reduced the mRNA expressions of IL-4,IL-10,GATA3 and FOXP3,and molecular docking simulation verified that the active components of WQY had excellent binding abilities with IL-4,IL-10,GATA3 and FOXP3 proteins.Conclusion:The present study demonstrated that WQY effectively relieved AD symptoms in mice,decreased the inflammatory factors levels,regulated the balance of CD4^(+)and CD8^(+)cells,and the mechanism may be associated with the suppression of Th2 and Treg cell immune responses.展开更多
文摘目的建立基于高分辨质谱、全球天然产物分子网络(global natural products social molecular networking,GNPS)和网络药理学的中药治疗共病物质基础研究策略,并对开心散治疗抑郁症-阿尔茨海默病的物质基础进行系统分析。方法采用超高效液相色谱-轨道阱串联质谱(ultra-performance liquid chromatography-orbitrap tandem mass spectrometry,UPLCOrbitrap-MS/MS)对开心散提取物及其血浆和脑组织暴露样品进行非靶向二级质谱分析。然后,利用GNPS平台,基于开心散提取物和血浆样品所得质谱数据构建分子网络,结合化合物的精确质量数、保留时间、碎片信息、质谱裂解规律,快速鉴定开心散血浆暴露成分。基于血浆暴露成分,结合脑组织样品的高分辨质谱数据,鉴定开心散入脑成分。结合脑组织暴露成分和网络药理学技术探究开心散治疗抑郁症-阿尔茨海默病共病物质基础。结果从血浆中鉴定出开心散暴露成分共34个,从脑组织中鉴定出开心散暴露成分共21个,并筛选出开心散中具有治疗抑郁症-阿尔茨海默病共病活性的成分共19个,涉及皂苷类成分9个、寡糖酯类成分9个、酮类成分1个。结论建立了中药共病物质基础快速发现的新策略,为中药“异病同治”研究提供方法支撑,且为开心散治疗抑郁症-阿尔茨海默病共病的物质基础阐释及其临床应用提供数据支撑。
基金supported by grants from the National Natural Science Foundation of China(82004252)the Project of Administration of Traditional Chinese Medicine of Guangdong Province(202405112017596500)the Basic and Applied Basic Research Foundation of Guangzhou Municipal Science and Technology Bureau(202102020533).
文摘Background:Wenqing Yin(WQY)is a classic prescription used to treat skin diseases like atopic dermatitis(AD)in China,and the aim of this study is to investigate the therapeutic effects and molecular mechanisms of WQY on AD.Methods:The DNFB-induced mouse models of AD were established to investigate the therapeutic effects of WQY on AD.The symptoms of AD in the ears and backs of the mice were assessed,while inflammatory factors in the ear were quantified using quantitative real-time-polymerase chain reaction(qRT-PCR),and the percentages of CD4^(+)and CD8^(+)cells in the spleen were analyzed through flow cytometry.The compounds in WQY were identified using ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)analysis and the key targets and pathways of WQY to treat AD were predicted by network pharmacology.Subsequently,the key genes were tested and verified by qRT-PCR,and the potential active components and target proteins were verified by molecular docking.Results:WQY relieved the AD symptoms and histopathological injuries in the ear and back skin of mice with AD.Meanwhile,WQY significantly reduced the levels of inflammatory factors IL-6 and IL-1βin ear tissue,as well as the ratio of CD4^(+)/CD8^(+)cells in spleen.Additionally,a total of 142 compounds were identified from the water extract of WQY by UPLC-Orbitrap-MS/MS.39 key targets related to AD were screened out by network pharmacology methods.The KEGG analysis indicated that the effects of WQY were primarily mediated through pathways associated with Toll-like receptor signaling and T cell receptor signaling.Moreover,the results of qRT-PCR demonstrated that WQY significantly reduced the mRNA expressions of IL-4,IL-10,GATA3 and FOXP3,and molecular docking simulation verified that the active components of WQY had excellent binding abilities with IL-4,IL-10,GATA3 and FOXP3 proteins.Conclusion:The present study demonstrated that WQY effectively relieved AD symptoms in mice,decreased the inflammatory factors levels,regulated the balance of CD4^(+)and CD8^(+)cells,and the mechanism may be associated with the suppression of Th2 and Treg cell immune responses.
文摘基于超高效液相色谱-四极杆静电场轨道阱质谱(UPLC-Q-Exactive Orbitrap-MS)技术及网络药理学探讨化浊散结除痹方治疗痛风性关节炎(gouty arthritis,GA)的药效物质及潜在机制。采用UPLC-Q-Exactive Orbitrap-MS技术鉴定化浊散结除痹方药物成分,对其有效成分进行定性分析,共鉴定出化浊散结除痹方中184个有效成分;通过PharmMapper在线数据库筛选有效成分靶点897个,在OMIM、GeneCards、CTD等数据库获取GA相关的疾病靶点491个,进行韦恩分析后获得二者的交集靶点60个,通过Cytoscape平台构建“成分靶点-GA靶点”网络图,利用STRING数据库构建蛋白-蛋白互作网络,筛选出16个核心靶点,将核心靶点进行基因本体论(Gene Ontology,GO)与京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)信号通路的富集分析,并构建“成分-靶点-通路”网络图,发现该方治疗GA的主要有效成分为酚类、黄酮类、生物碱类、萜类化合物,关键靶点有SRC、MMP3、MMP9、REN、ALB、IGF1R、PPARG、MAPK1、HPRT1、CASP1,通过GO分析发现其治疗GA主要涉及脂质反应、细菌反应、生物刺激的反应等生物过程,通过KEGG分析发现其治疗GA相关的通路有脂质和动脉粥样硬化、中性粒细胞胞外诱捕网、IL-17等。综上,该研究揭示了酚类、黄酮类、生物碱类、萜类化合物可能是化浊散结除痹方治疗GA的核心药效物质,其药效机制可能与SRC、MMP3、MMP9等靶点及脂质和动脉粥样硬化、中性粒细胞胞外诱捕网、IL-17等通路相关。
