目的 基于中医药整合药理学和转录组学探究丹栀逍遥散防治焦虑肝郁化火证的作用机制。方法 采用中医药整合药理学研究平台v2.0，检索并获取丹栀逍遥散的活性成分及靶标、焦虑肝郁化火证靶标信息，构建中药-成分-靶点-通路网络。将交集基因导入String数据库，构建交集靶点的蛋白互作网络，以确定最终需要验证的靶标。利用GEO数据库转录组学验证整合网络药理学结果，同时采用动物实验进行验证。结果 共筛选出与焦虑肝郁化火证相关的靶标13个，丹栀逍遥散活性成分430种。整合药理学结果显示，丹栀逍遥散防治焦虑肝郁化火证主要通过调控α-氨基-3-羟基-5-甲基-4-异噁唑丙酸受体（α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor，AMPAR）、蛋白激酶A（protein kinase A，PKA）、N-甲基-D-天冬氨酸（N-methyl-D-aspartic acid，NMDA）受体的激活及磷脂酰肌醇3-激酶（phosphatidylinositide 3-kinases，PI3K）/蛋白激酶B（protein kinase B，Akt）等信号通路。GEO微阵列芯片GSE4035数据集转录组学关键靶标雌激素受体α（estrogen receptor 1，ESR1）、腺苷酸环化酶5（adenylate cyclase 5，ADCY5）、谷氨酸离子型受体NMDA型亚基2A（glutamate ionotropic receptor NMDA type subunit 2A，GRIN2A）基因为差异表达基因（differential gene expression，DEGs）。实时荧光定量聚合酶链反应（quantitative real-time polymerase chain reaction，qRT-PCR）结果显示，ADCY5 mRNA表达上调和GRIN2A mRNA表达下调。结论 丹栀逍遥散可能是通过上调ADCY5 mRNA表达和下调GRIN2A mRNA表达进而调节脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）与其受体的结合和谷氨酸神经传递介导，进一步改善神经元生长、突触传导与突触的可塑性，从而发挥其生物学效应。

Objective To explore the mechanism of Danzhi Xiaoyao Powder (丹栀逍遥散) in the prevention and treatment of anxiety with liver depression transforming into fire syndrome based on integrated pharmacology of traditional Chinese medicine and transcriptomics. Methods The integrated pharmacology research platform of traditional Chinese medicine v2.0 was used to retrieve and obtain the active ingredients and target information of Danzhi Xiaoyao Powder, and the target information of anxiety with liver depression transforming into fire syndrome, and construct the network of traditional Chinese medicine-component-target-pathway. The intersection genes were imported into the String database, and the protein interaction network of intersection targets was constructed to determine the target that needed to be verified. The transcriptomic of GEO database were used to verify the integrated network pharmacology results, and animal experiments were used to verify the results. Results A total of 13 targets related to anxiety with liver depression transforming into fire syndrome and 430 kinds of active ingredients of Danzhi Xiaoyao Powder were screened out, and the integrated pharmacological results showed that Danzhi Xiaoyao Powder prevented and treated anxiety with liver depression transforming into fire syndrome by mainly regulating the activation of α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR), protein kinase A (PKA), N-methyl-D-aspartic acid (NMDA) receptor, phosphatidylinositide 3-kinases (PI3K) of protein kinase B (Akt) and other signal pathways of anxiety liver depression and fire syndrome. The GEO microarray GSE4035 data set, the key targets of transcriptomics estrogen receptor 1 (ESR1), adenylate cyclase 5 (ADCY5) and glutamate ionotropic receptor NMDA type subunit 2A (GRIN2A) genes were differential gene expression (DEGs). Quantitative real-time polymerase chain reaction (qRT-PCR) results showed that ADCY5 mRNA expression was up-regulated and GRIN2A mRNA expression was down-regulated. Conclusion Danzhi Xiaoxiao Powder may regulate brain-derived neurotrophic factor (BDNF) by up-regulating the expression of ADCY5 mRNA and down-regulating the expression of GRIN2A mRNA, thus regulating the binding of BDNF and its receptors and the neurotransmission of glutamate, further improve neuron growth, synaptic conduction and synaptic plasticity, so as to exert its biological effects.

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国家自然科学基金项目（81703964）；云南省科技厅科技计划项目（202101AZ070001-021）；云南省教育厅基金项目：基于GC-NLRP3炎症小体通路研究丹栀逍遥散干预肝郁化火型焦虑行为大鼠的作用机制（2022Y332）；云南省科技厅科技人才与平台计划—云南省中医药学分子生物学重点实验室[2019DG016BK-基于整合网络药理学探究丹栀逍遥散防治郁化火型焦虑行为的作用机制（30172110174），BK-丹栀逍遥散通过GC-NLRP3信号通路干预焦虑行为大鼠的机制研究（30172110179）]