有毒乌头类中药的毒效网络整合分析：思路与方法

中药的有效性和安全性不仅是中医药传承和发展的坚实基础,也是中医药服务于世界人民健康事业的根本前提。因此多学科交叉融合、整合分析中药毒效的物质基础、作用原理、毒效关系,揭示"有故无殒、中病即止"的科学内涵,是中药进入国际医药市场亟待解决的关键问题。利用复杂生物网络研究中药系统药理学和系统毒理学,构建可靠的中药药效物质作用靶标相关的分子效应和毒理学性质网络,反映特定疾病状态的相关生物实体的分子网络及毒性机制,整合效应物质作用靶标网络与疾病生物网络,通过网络拓扑学和动力学等方法分析"药物-靶点-疾病"网络和"药物-靶点-活性/毒性"网络中有效物质与疾病、成分与毒性的相互作用关系,检识出复杂网络模型中的靶点组合、通路组合、子网组合、药物组合等,大大提高了中药复杂药效物质的研究效率。作者主要基于其研究团队近年来开展的乌头类中药研究工作,重点介绍开展有毒中药的毒效网络整合分析研究的思路和方法。

从临床多组学整合调控网络探究雷公藤多苷片治疗类风湿关节炎的“效-毒”关联机制

目的:通过雷公藤多苷片治疗类风湿关节炎疗效应答和诱发药源性肝损伤相关分子互作网络挖掘,揭示雷公藤多苷片的“效-毒”关联机制。方法:利用前期已获得临床来源的雷公藤多苷片疗效应答相关基因表达谱和诱发药源性肝损伤相关蛋白质表达谱数据,根据生物分子间的相互作用信息,构建雷公藤多苷片“效-毒”关联网络,并通过计算网络节点的拓扑特征值,筛选关键网络靶标和挖掘其生物学功能。结果:经临床转录组学检测,获得雷公藤多苷片治疗类风湿关节炎疗效相关差异表达基因119个。经临床蛋白质组学检测,获得雷公藤多苷片诱发药源性肝损伤相关差异表达蛋白质49个。进一步的临床症状富集分析表明,雷公藤多苷片治疗类风湿关节炎的效应基因与痹症相关中医症状和医学临床表型紧密相关,雷公藤多苷片诱发药源性肝损伤相关的差异蛋白质最显著富集于消化系统异常的临床症状,可见,上述多组学数据能够表征雷公藤多苷片治疗类风湿关节炎和诱发药源性肝损伤的主要临床症状。进而,“效-毒”关联网络挖掘表明,雷公藤多苷片治疗类风湿关节炎的关键效应靶标和诱发药源性肝损伤核心致毒蛋白质均参与机体“免疫-炎症”失衡的调节,特别在中性粒细胞脱颗粒、补体级联反应、蛋白质翻译后修饰介导的免疫炎症反应等方面发挥着重要作用;二者还显著富集于肾素-血管紧张素系统(RAS)和丝裂原活化蛋白激酶(MAPK)信号通路等与细胞增殖和细胞周期调控密切相关的信号通路,以及参与糖原代谢和氧化还原反应等肝脏功能直接相关的生物学通路。结论:本研究采用临床表型组学、临床转录组学和蛋白质组学整合的研究策略,系统阐释了雷公藤多苷片“效-毒”的关联特征和分子机制,为探索雷公藤多苷片“增效减毒”机制奠定良好的数据基础。

四逆汤配伍环境下的附子“效-毒网络”交集调控研究

目的:采用网络药理学方法探索四逆汤配伍环境下,甘草、干姜对附子作用基因"效-毒网络"交集调控的机制。方法:应用Cytoscape软件和插件Agilent Literature Search,进行文本挖掘并建立附子、甘草、干姜作用的基因相互关系网络;根据附子抗心衰、神经毒性和心脏毒性形成附子"效-毒网络"交集;应用插件Clusterviz进行靶标基因聚类,从DAVID数据库中对甘草、干姜调控附子"效-毒网络"交集的可能通路进行预测。结果:与附子神经毒性、心脏毒性和抗心衰作用同时相关的基因有5个,AKT1,BAX,HCC,IL6,IL8,形成附子"效-毒网络"交集有47个节点基因;甘草、干姜与附子"效-毒网络"交集的重合基因分别有29个和27个,可能调控通路有23条和17条。结论:在四逆汤配伍环境下,甘草、干姜可能通过影响免疫-炎症反应信号通路、细胞凋亡-自噬信号通路、神经细胞与心肌细胞缺血缺氧保护信号通路等途径调控附子的效/毒效应。

Study on the compatibility principle of Wutou Decoction based on network pharmacology

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.

中药毒性的研究现状及其超分子“印迹模板”调控机制的探讨

基于前期提出的中医药超分子“印迹模板”自主作用规律,分析中药毒性的研究现状,结合超分子“气析”理论,阐述中药毒性的超分子“印迹模板”的调控机制:中药(客体超分子)可与人体(主体超分子)发生作用,中药成分群与机体内对应的孔穴通道进行识别,有毒中药的量效问题、减毒处理都与平衡常数及浓度密切相关,当具有相同(似)“印迹模板”药物的浓度相同时,毒性的大小取决于其与靶点作用的平衡常数。因此,提出了中药毒性的平衡常数研究思路,可用超分子印迹作用的平衡常数表征中药毒性,通过研究平衡常数的大小可判断成分产生毒性的主客体分子间作用程度,揭示毒性效应的主客体超分子间相互作用,促进中医药超分子体系及中药毒性的研究。