摘要
Objective To elucidate the mechanisms underlying the therapeutic effects of Fufang Ejiao Jiang(复方阿胶浆,FFEJJ)on aplastic anemia(AA)using integrated network pharmacology and serum metabolomics.Methods Traditional Chinese Medicine Systems Pharmacology(TCMSP),Pubmed,integrative pharmacology-based research platform of traditional Chinese medicine(TCMIP),and Bioinformatics Analysis Tool for Molecular mech ANism of Traditional Chinese Medicine(BATMAN-TCM)were used to identify the constituents and putative targets of FFEJJ.Gene Cards and DisGeNET databases were used to identify AA-associated targets.We constructed a herb-component-target network and analyzed the protein-protein interaction(PPI)network.Potential mechanisms were determined using Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.In addition,an AA model was established using acetylphenylhydrazine(APH)and cetylphenylhydrazine(CTX).Ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS)-based serum metabolomics was applied to screen potential metabolites and the related pathways associated with AA and the potential anti-anemic effects of FFEJJ.Results A total of 30 active components of FFEJJ and 24 targets were related to AA.PPI network analysis showed that VEGFA,AKT1,IL-6,CASP3,and ICAM1 were key nodes overlapping with proteins known to be related to AA.KEGG pathway enrichment analysis revealed that the presumed targets of FFEJJ were mainly associated with pathways linked to the promotion of hematopoiesis and improvement of the hematopoietic microenvironment.A total of 423 metabolite biomarkers were identified between the control and AA models,which are involved in the development of AA.In contrast,FFEJJ reversed the 79 differential metabolites altered by AA.Pathway analysis suggested that the synergistic effects of FFEJJ were mainly enriched in 24 metabolic pathways.Among them,sphingolipid metabolism,glycerophospholipid metabolism,and arachidonic acid metabolism were related to promoting hematopoiesis and improving the hematopoietic microenvironment,which partially conforms with network pharmacology.The interaction network formed by three key differential metabolites,including hydroxy-eicosatetraenoic acid(HETE),sphingosine 1-phosphate(S1 P),and lysophosphatidylcholine(lyso PC),and three predicted network targets(VEGFA,CASP3,and ICAM1)may be the potential mechanism underlying the anti-AA action of the multi-component of FFEJJ.Conclusion FFEJJ could be an alternative treatment option for AA.It acts by promoting hematopoiesis and improving the hematopoietic microenvironment.Network pharmacology-integrated metabolomics makes it possible to analyze TCMs from a systems perspective and at the molecular level.
目的采用网络药理学和血清代谢组学相结合的方法,探讨复方阿胶浆(FFEJJ)治疗再生障碍性贫血(AA)潜在作用机制。方法从中药系统药理学数据库与分析平台(TCMSP),Pubmed,中医药综合药理研究平台(TCMIP),Bioinformatics,中药分子机制的生物信息学分析工具(BATMAN-TCM)用于确定FFEJJ的成分和假定的靶点。使用GeneCard和DisGeNET数据库获得AA相关的靶点,构建草药-成分-靶点网络,并分析蛋白质相互作用(PPI)网络,通过京都基因和基因组百科全书(KEGG)对潜在途径进行富集分析。此外,联合乙酰苯肼(APH)和环磷酰胺(CTX)建立AA模型评估FFEJJ的血液保护作用,基于超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF/MS)的血清代谢组学筛选FFEJJ参与抗贫血的潜在代谢标志物和相关代谢途径。结果共筛选30个FFEJJ活性成分和24个与AA调控相关的靶点。已报道的PPI网络分析显示VEGFA、AKT1、IL-6、CASP3和ICAM1与AA发生发展相关的关键靶点。KEGG通路富集分析表明,FFEJJ的假定靶点主要涉及与促进造血和改善造血微环境相关的途径。在对照组和AA模型之间鉴定出423个代谢物生物标记物,它们参与了AA的发展,而FFEJJ逆转了79种AA改变的差异代谢物。KEGG通路分析表明,FFEJJ的协同效应主要集中在24条代谢途径中,其中,鞘脂代谢、甘油磷脂代谢和花生四烯酸代谢与促进造血和改善造血微环境有关,这与网络药理分析结果一致。3个关键差异代谢物,羟基二十碳四烯酸(HETE)、1-磷酸鞘氨醇(S1P)和溶血磷脂酰胆碱(lysoPC)与3个网络预测靶点VEGFA、CASP3和ICAM1形成交互网络可能是FFEJJ多组分抗AA作用的潜在机制。结论FFEJJ是治疗AA的一种替代方法,其潜在机制是促进造血和改善造血微环境。本文研究策略提示网络药理学整合代谢组学研究使从系统角度和分子水平阐明中医药作用成为可能。
基金
funding support from the Natural Science Foundation of China(No.81673585,No.81874493,No.81573956)
Program of Survey of Chinese Medicines of China(No.[2017]66)
Science Foundation of Hunan Province(No.2019JJ50345,No.2020JJ5325,No.2021168)
Key Research and Development Project of Changsha Science and Technology(No.kq1901067)
Training Program for Excellent Young Innovators of Changsha(No.kq1802017)
Research on the Comprehensive Development and Utilization of Characteristic Traditional Chinese Medicine Resources(No.2060302)
the Support of Hunan Province Traditional Chinese Medicine Preparation and Quality Traceability Engineering and Technology Center
the 2011 Collaboration and Innovation Center for Digital Chinese Medicine in Hunan。
