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Abstract：
Background: Mesaconitine (MA), a diester-diterpenoid alkaloid extracted from the medicinal herb Aconitum carmichaelii, is commonly used to treat various diseases. Previous studies have indicated the potent toxicity of aconitum despite its pharmacological activities, with limited understanding of its effects on the nervous system and the underlying mechanisms. Methods: HT22 cells and zebrafish were used to investigate the neurotoxic effects of MA both in vitro and in vivo, employing multi-omics techniques to explore the potential mechanisms of toxicity. Results: Our results demonstrated that treatment with MA induces neurotoxicity in zebrafish and HT22 cells. Subsequent analysis revealed that MA induced oxidative stress, as well as structural and functional damage to mitochondria in HT22 cells, accompanied by an upregulation of mRNA and protein expression related to autophagic and lysosomal pathways. Furthermore, methylated RNA immunoprecipitation sequencing (MeRIP-seq) showed a correlation between the expression of autophagy-related genes and N6-methyladenosine (m6A) modification following MA treatment. In addition, we identified METTL14 as a potential regulator of m6A methylation in HT22 cells after exposure to MA. Conclusion: Our study has contributed to a thorough mechanistic elucidation of the neurotoxic effects caused by MA, and has provided valuable insights for optimizing the rational utilization of traditional Chinese medicine formulations containing aconitum in clinical practice.
摘要：
背景：中乌头碱（MA），从草药乌头中提取的二酯-二萜生物碱，通常用于治疗各种疾病。先前的研究表明，尽管乌头具有药理活性，但其具有强大的毒性，对其对神经系统和潜在机制的影响了解有限。方法:用HT22细胞和斑马鱼研究MA在体外和体内的神经毒性作用。采用多组学技术探索潜在的毒性机制。结果：我们的结果表明，用MA处理可诱导斑马鱼和HT22细胞的神经毒性。随后的分析表明，MA诱导氧化应激，以及HT22细胞中线粒体的结构和功能损伤，伴随着与自噬和溶酶体途径相关的mRNA和蛋白质表达的上调。此外，甲基化RNA免疫沉淀测序（MeRIP-seq）显示自噬相关基因的表达与MA处理后的N6-甲基腺苷（m6A）修饰之间存在相关性。此外,我们确定METTL14是暴露于MA后HT22细胞中m6A甲基化的潜在调节因子。结论：我们的研究有助于对MA引起的神经毒性作用进行彻底的机制阐明，为临床上优化含乌头中药配方的合理使用提供了有价值的见解。
