Qi Xue-Jia, Huang Chong-Yin, Zuo Meng-Ting, Gong Meng-Die, Huang Si-Juan, Tang Mo-Huan, Liu Zhao-Ying
College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China.
Hunan Engineering Technology Research Center of Veterinary Drugs, Hunan Agricultural University, Changsha 410128, China.
Metabolites. 2023 Jan 28;13(2):195. doi: 10.3390/metabo13020195.
is a medicinal plant that has been used to treat various diseases, but it is also well-known for its high toxicity. Complex alkaloids are considered the main poisonous components in . However, the toxic mechanism of remains ambiguous. In this work, network pharmacology and experimental verification were combined to systematically explore the specific mechanism of toxicity. The alkaloid compounds and candidate targets of , as well as related targets of excitotoxicity, were collected from public databases. The crucial targets were determined by constructing a protein-protein interaction (PPI) network. Subsequently, Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to explore the bioprocesses and signaling pathways involved in the excitotoxicity corresponding to alkaloids in . Then, the binding affinity between the main poisonous alkaloids and key targets was verified by molecular docking. Finally, animal experiments were conducted to further evaluate the potential mechanisms of toxicity. A total of 85 alkaloids in associated with 214 excitotoxicity-related targets were predicted by network pharmacology. Functional analysis showed that the toxicity of was mainly related to the protein phosphorylation reaction and plasma membrane function. There were also 164 pathways involved in the toxic mechanism, such as the calcium signaling pathway and MAPK signaling pathway. Molecular docking showed that alkaloids have high affinity with core targets, including MAPK3, SRC, MAPK1, NMDAR and NMDAR. In addition, the difference of binding affinity may be the basis of toxicity differences among different alkaloids. Humantenirine showed significant sex differences, and the LD values of female and male mice were 0.071 mg·kg and 0.149 mg·kg, respectively. Furthermore, we found that N-methyl-D-aspartic acid (NMDA), a specific NMDA receptor agonist, could significantly increase the survival rate of acute humantenirine-poisoned mice. The results also show that humantenirine could upregulate the phosphorylation level of MAPK3/1 and decrease ATP content and mitochondrial membrane potential in hippocampal tissue, while NMDA could rescue humantenirine-induced excitotoxicity by restoring the function of mitochondria. This study revealed the toxic components and potential toxic mechanism of . These findings provide a theoretical basis for further study of the toxic mechanism of and potential therapeutic strategies for poisoning.
是一种用于治疗多种疾病的药用植物，但它也因其高毒性而闻名。复杂生物碱被认为是其主要有毒成分。然而，其毒性机制仍不明确。在这项工作中，将网络药理学与实验验证相结合，系统地探索其毒性的具体机制。从公共数据库收集其生物碱化合物和候选靶点，以及兴奋性毒性的相关靶点。通过构建蛋白质-蛋白质相互作用（PPI）网络确定关键靶点。随后，利用基因本体论（GO）和京都基因与基因组百科全书（KEGG）探索与其中生物碱对应的兴奋性毒性所涉及的生物过程和信号通路。然后，通过分子对接验证主要有毒生物碱与关键靶点之间的结合亲和力。最后，进行动物实验以进一步评估其毒性的潜在机制。网络药理学预测其共有85种生物碱与214个兴奋性毒性相关靶点有关。功能分析表明，其毒性主要与蛋白质磷酸化反应和质膜功能有关。其毒性机制还涉及164条信号通路，如钙信号通路和丝裂原活化蛋白激酶（MAPK）信号通路。分子对接表明，生物碱与核心靶点具有高亲和力，包括MAPK3、SRC、MAPK1、N-甲基-D-天冬氨酸受体（NMDAR）等。此外，结合亲和力的差异可能是不同生物碱毒性差异的基础。胡蔓藤碱乙表现出显著的性别差异，雌性和雄性小鼠的半数致死量（LD）值分别为0.071 mg·kg和0.149 mg·kg。此外，我们发现N-甲基-D-天冬氨酸（NMDA），一种特异性NMDAR激动剂，可显著提高急性胡蔓藤碱乙中毒小鼠的存活率。结果还表明，胡蔓藤碱乙可上调海马组织中MAPK3/1的磷酸化水平，降低ATP含量和线粒体膜电位，而NMDA可通过恢复线粒体功能来挽救胡蔓藤碱乙诱导的兴奋性毒性。本研究揭示了其有毒成分和潜在毒性机制。这些发现为进一步研究其毒性机制及胡蔓藤碱中毒的潜在治疗策略提供了理论依据。
