基于网络药理学与分子对接技术的石菖蒲治疗癫痫作用机制研究＊

目的 利用网络药理学和分子对接技术研究石菖蒲抗癫痫的有效成分及作用机制。方法 运用中药系统药理数据库及分析平台检索石菖蒲有效成分及主要靶点，通过GeneCards数据库收集癫痫疾病的相关潜在作用靶点，运用Cytoscape.3.7.1和 String绘制石菖蒲-癫痫靶点互作关系图，运用Metascape数据库进行GO和KEGG信号通路富集分析。运用AutoDock Vina 1.1.2软件对石菖蒲主要有效成分与癫痫相关靶标进行分子对接验证；通过构建戊四唑诱导的癫痫模型初步评价石菖蒲提取物及潜在有效成分的抗癫痫作用。结果 由 TCMSP 筛选得到4个有效成分及76个主要靶点。癫痫疾病以“Score”大于1.5分进行筛选得到3685个靶标。GO富集分析得到BP条目10个，细胞CC条目9个，MF条目10个（P<0.01），主要涉及核受体活性、离子通道活性、激素结合、神经递质受体活性、蛋白激酶活性、钙调蛋白结合等。KEGG 富集得到14条通路，主要涉及c型凝集素受体信号通路、雌激素信号通路、Ca²⁺信号通路、逆行内源性大麻素信号通路等。分子对接结果显示，桉脂素、山奈酚和8-异戊烯基山奈酚与癫痫关键靶标GABRA2、PPARG等具有良好的结合活性。动物实验结果表明，石菖蒲水提取物及主要潜在有效成分桉脂素和山奈酚在PTZ致小鼠癫痫模型上显示出较好的抗癫痫活性。结论 本研究通过动物实验验证了石菖蒲提取物和潜在有效成分的抗癫痫活性，并且运用网络药理学和分子对接技术探讨了其抗癫痫作用机制，发现石菖蒲可通过多个成分如桉脂素、山奈酚、8-异戊烯基山奈酚，多靶点如GABA_A、PPARG等，多条通路如雌激素信号通路、钙信号通路对机体产生协调效应从而抑制癫痫的发作。该研究提示石菖蒲治疗癫痫具有多成分、多靶点、多通路特点，为石菖蒲的进一步研究提供数据支撑。

Investigating the Pharmacological Mechanism of Rhizoma Acori Tatarinowii Acting on Epilepsy via Network Pharmacology and Molecular docking

Objective To explore the effective ingredients and mechanism of Rhizoma Acori Tatarinowii in the therapy of epilepsy based on network pharmacology.Methods The Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) was used to filter the effective ingredients of Rhizoma Acori Tatarinowii and collect the main targets. We used Cytoscape.3.7.1 and String software to construct Rhizoma Acori Tatarinowii-epilepsy targets interaction diagram. The common target of both drugs and disease was introduced into Metascape for GO biological function analysis and KEGG pathway analysis. AutoDock Vina 1.1.2 software was used to verify the molecular docking validation between the main active components of Rhizoma Acori Tatarinowii and epilepsy related targets. The anticonvulsant activity of Rhizoma Acori Tatarinowii aqueous extract and its bioactive compounds eudesmin and kaempferol was assessed using the PTZ-induced seizures.Results Four active ingredients and 76 significant targets of Rhizoma Acori Tatarinowii were screened by TCMSP. The epilepsy disease was screened with a "score" greater than 1.5. Therefore, it was obtained 3685 main targets. A total of 10 biological process (BP) entries, 9 cellular component (CC) entries, and 10 molecular function (MF) entries were obtained by GO enrichment analysis, mainly including nuclear receptor activity, channel activity, hormone binding, neurotransmitter receptor activity, protein kinase activity, calcium modulin binding, etc. There were 14 KEGG pathways, including c-type lectin receptor (CLR) signaling pathway, estrogen signaling pathway, calcium signaling pathway, Retrograde end cannabinoid signaling pathway, etc. Molecular docking results showed that kaempferol, 8-isopentylkaempferol, eudesmin had good binding activity with key epileptic targets GABRA2, and PPARG. Animal experiment results confirmed that Rhizoma Acori Tatarinowii aqueous extract as well as eudesmin and kaempferol delayed the onset of seizures and reduced mortality in mice.Conclusion In this study, the antiepileptic activities of Rhizoma Acori Tatarinowii and its potential active components are verified by animal experiments, and the antiepileptic mechanism is discussed based on network pharmacology and molecular docking technology. It confirms that Rhizoma Acori Tatarinowii can induce coordinated effects on the body through multiple components such as kaempferol, 8-isopentyl kaempferol, eudesmin, multi-target sites such as GABA_A and PPARG, etc., multi-pathway such as estrogen signaling pathway and calcium signaling pathway to inhibit the seizure of epilepsy. This study shows that Rhizoma Acori Tatarinowii treating on epilepsy by the multi-component, multi-target and multi-channel, which provides data support for the further study of Rhizoma Acori Tatarinowii.