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目标探讨乌头汤中川乌与黄芪的增效作用及甘草联合川乌减毒作用的机制,阐明乌头汤的配伍原则.方法通过中药系统药理学数据库及分析平台(TCMSP)、中医药综合数据库(TCMID)进行相关药物的活性成分及靶点的检索.针对川乌毒性靶点的研究,我们主要选取了其成分中5种具有明确毒性的成分并运用瑞士的靶点预测平台(SwissTargetPrediction)进行相关靶点收集;通过基因疾病关联数据库(DisGeNET),人类基因数据库(GenCards)和人类在线孟德尔遗传(OMIM)数据库来进行类风湿关节炎(RA)疾病相关靶点的整理.我们将通过上述获得的药物靶点与疾病靶点取交集,并定义相关交集靶点为潜在治疗靶点.运用可视化复杂网络(Cytoscape 3.9.0)软件实现药物成分-靶点网络的构建.使用注释、可视化和集成发现的数据库(DAV-ID)平台来完成相关基因本体(GO)富集分析及京都基因和基因组百科全书(KEGG)通路分析.结果共得到191个甘草相关的靶点,171个黄芪的靶点及103个川乌靶点(川乌中有效成分次乌头碱的靶点获得主要通过相关文献查找及SwissTargetPrediction平台).同时,我们共获得了5872个与RA发病具有相关性的靶点.经过筛选后,我们共获得了13个增效相关的靶点及9个减毒相关的靶点.其中以PGR为核心发挥增效作用靶点,KCNH2为核心发挥减毒作用的靶点.最终,对于增效相关靶点的GO分析有23个模块,其中显示RNA聚合酶II启动子转录的正调节、类固醇激素介导的信号通路、质膜和蛋白质结合(P<?0.01)等可能与增效作用的实现密切相关;同时,KEGG通路分析共得到13条与增效作用密切相关的通路[如雌激素、胆碱能突触和磷酸肌醇3-激酶/蛋白激酶B(PI3K-Akt)信号通路等].有关减毒靶点的GO分析有28个模块,显示与G-蛋白偶联受体、质膜和药物结合等生物过程密切相关;KEGG通路分析共得到5条与减毒作用密切相关的通路:胆碱能突触通路、钙离子信号通路、肌动蛋白细胞骨架的调节、神经活性配体-受体相互作用以及含血清素的神经突触.结论乌头汤中川乌及黄芪主要通过雌激素信号通路、PI3K/Akt信号通路以调节PGR基因为核心发挥增效作用.同时,川乌与甘草则通过胆碱能突触、钙离子信号通路以KCNH2基因为核心发挥减毒作用.“,”Objective To investigate the underlying drug enhancement mechanisms of the Chuanwu (Aconiti Radix) and Huangqi (Astragali Radix) combination and toxicity reduction of Chuan-wu combined with Gancao (Glycyrrhizae Radix et Rhizoma) in Wutou Decoction (乌头汤, WTD), and to elucidate the compatibility principle.Methods The active compounds and potential effective targets of the selected combinations were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Traditional Chinese Medicines Integrated Database (TCMID). The toxicity of Chuanwu (Aconiti Radix) was investigated by selecting all five toxic compounds from the literature and the TCMSP database, and obtaining their targets through SwissTargetPrediction. Targets related to rheumatoid arthritis (RA) were searched using Dis-GeNET, GenCards, and Online Mendelian Inheritance in Man (OMIM). Mutual targets between the drug pairs and RA were selected as potential RA therapy targets. The medicinally active compound-target network was constructed using Cytoscape 3.9.0. Gene ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrich-ment were performed using the Database for Annotation, Visualization, and Integrated Dis-covery (DAVID) platform. Results We obtained 191 active compound targets for Gancao (Glycyrrhizae Radix et Rhizoma), 171 for Huangqi (Astragali Radix), and 103 for Chuanwu (Radix Aconiti) (hypo-aconitine\'s target was obtained through literature and SwissTargetPrediction). A total of 5872 genes were obtained for RA. A drug-active compound-target network involving 13 effect-en-hancing and nine toxicity reduction targets was constructed. PGR was the main effect en-hancement target, and KCNH2 was the main toxicity reduction target. The effect-enhancing targets were related to 23 GO terms (such as positive regulation of transcription from RNA polymerase Ⅱ promoter, steroid hormone-mediated signaling pathway, plasma membrane, and protein binding) (P < 0.01), and 13 KEGG pathways related to synergism [such as estro-gen signaling pathway, cholinergic synapse, and phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway]. The toxicity reduction targets were related to 28 GO terms (mainly involes G-protein coupled receptor signaling pathway, plasma membrane, and drug binding) (P < 0.01), and five KEGG pathways related to toxicity reduction (cholinergic syn-apse, calcium signaling pathway, regulation of actin cytoskeleton, neuroactive ligand-recept-or interaction, and serotonergic synapse).Conclusion The combination of Chuanwu (Aconiti Radix) and Huangqi (Astragali Radix) plays an important effect-enhancing role in WTD and involves the estrogen and PI3K/Akt sig-naling pathways, with PGR as the core. The Chuanwu (Aconiti Radix) and Gancao (Gly-cyrrhizae Radix et Rhizoma) combination decreases toxicity in WTD and is associated with the cholinergic synapse and calcium signaling pathways, with KCNH2 as the core.