目的 运用网络毒理学与分子对接技术探究乌头碱引发神经毒性的作用机制。方法 通过Swiss Target Prediction、PharmMapper以及CTD数据库系统收集乌头碱的潜在作用靶点。利用CTD、PharmGKB、OMIM数据库检索神经毒性相关基因靶点，将乌头碱对应靶点与神经毒性相关靶点取交集，确定候选靶点。基于STRING数据库平台构建候选靶点的蛋白质相互作用（PPI）网络，并通过CytoScape 3.9.1软件进行网络拓扑学分析，构建核心靶点网络图。通过Metascape在线分析平台，对候选靶点进行基因本体（GO）、京都基因与基因组百科全书（KEGG）富集分析。采用PyMOL分子可视化软件实施分子对接模拟，对网络毒理学预测结果进行验证。结果 得到乌头碱对应靶点848个，神经毒性相关靶点1 713个，交集靶点为144个，其中核心靶点20个，包括蛋白激酶B1（Akt1）、白蛋白（ALB）、胱天蛋白酶3（CASP3）、白细胞介素-6（IL-6）、肿瘤坏死因子（TNF）等；GO富集分析显示乌头碱致神经毒性主要涉及外源物质刺激反应和外源物代谢过程等生物过程，线粒体膜和突触后部等细胞组分，以及氧化还原酶活性和转录因子结合等分子功能；KEGG分析主要富集于糖尿病并发症中的晚期糖基化终产物及其受体（AGE-RAGE）、神经营养因子、催乳素等信号通路。分子对接结果显示乌头碱与排名前10位的核心靶点结合能均小于−5.0 kcal/mol，具有良好的结合能力。结论 乌头碱产生神经毒性具有多靶点、多途径的作用规律，通过调控TNF、CASP3、Akt1等靶点影响AGE-RAGE、神经营养因子、催乳素等信号通路从而引发神经毒性。

Objective To investigate the neurotoxic mechanism induced by aconitine using network toxicology and molecular docking techniques. Methods The potential targets of aconitine were systematically collected through the Swiss Target Prediction, PharmMapper, and CTD databases. Neurotoxicity-related gene targets were retrieved using databases such as CTD, PharmGKB, OMIM. The targets of aconitine and neurotoxicity-related targets were intersected to identify candidate targets. A PPI network of candidate targets was constructed based on the STRING database platform, and network topological analysis was performed using CytoScape 3.9.1 software to build a core target network diagram. GO and KEGG enrichment analyses of candidate targets were conducted via the Metascape online analysis platform. Molecular docking simulations were implemented using PyMOL molecular visualization software to conduct preliminary verification of the network toxicology prediction results. Results A total of 848 potential targets of aconitine and 1 713 targets associated with neurotoxicity were identified, yielding 144 intersecting targets. Among these, 20 core targets were determined, including Akt1, ALB, CASP3, IL-6, TNF, and others. GO enrichment analysis revealed that the neurotoxicity induced by aconitine is primarily involved in biological processes such as the response to xenobiotic stimulus and xenobiotic metabolic process, cellular components including the mitochondrial membrane and postsynapse, and molecular functions like oxidoreductase activity and transcription factor binding. KEGG pathway analysis indicated significant enrichment in signaling pathways related to the AGE-RAGE axis, neurotrophin, and prolactin signaling. Molecular docking results demonstrated that the binding energies between aconitine and the top 10 core targets were all lower than -5.0 kcal/mol, indicating favorable binding affinity. Conclusion Aconitine induces neurotoxicity through multi-target and multi-pathway mechanisms, potentially by regulating targets such as TNF, CASP3, and Akt1, thereby affecting signaling pathways including AGE-RAGE, neurotrophin, and prolactin, ultimately leading to neurotoxic effects.

R965;R971

陈可冀中西医结合发展基金项目（CKJ2023002）