目的 基于斑马鱼幼鱼模型探究士的宁的肝毒性及潜在作用机制。方法 通过急性毒性实验确定士的宁的10%致死浓度（10% lethal concentration，LC₁₀），以0.33、0.66μg/mL（1/4 LC₁₀、1/2 LC₁₀）2个亚致死质量浓度干预斑马鱼幼鱼，通过体视显微成像观察肝脏表型变化，吖啶橙染色检测肝细胞凋亡，组织病理学分析肝组织空泡化程度，并测定丙氨酸氨基转移酶（alanine aminotransferase，ALT）和天冬氨酸氨基转移酶（aspartate aminotransferase，AST）活性。进一步采用转录组测序技术筛选差异表达基因，并通过qRT-PCR验证关键基因表达水平。结果 士的宁亚致死剂量暴露诱导斑马鱼幼鱼肝脏形态异常，病理切片显示肝细胞空泡化，ALT和AST活性均显著升高（P＜0.05、0.01）。转录组分析发现脂质代谢通路被显著富集，其中细胞死亡诱导DFF45样效应因子B（cell death-inducing DFF45-like effector B，CIDEB）、细胞色素P450家族7亚家族A成员1（cytochrome P450 family 7 subfamily A member 1，CYP7A1）、成纤维细胞生长因子19（fibroblast growth factor 19，FGF19）等关键调控基因表达异常。结论 士的宁具有明显肝毒性，其毒性作用机制可能涉及调控脂质代谢通路关键基因（如CIDEB、CYP7A1、FGF19）的异常表达，进而引发脂质代谢紊乱，为马钱子及含士的宁相关药物制剂的临床安全合理用药提供理论参考。

Objective To investigate the hepatotoxicity and potential mechanism of strychnine based on zebrafish larvae model. Methods The acute toxicity test was performed to determine the 10% lethal concentration (LC₁₀) of strychnine, and then the sub-lethal concentrations of 0.33, 0.66 μg/mL (1/4 LC₁₀, 1/2 LC₁₀) were given to zebrafish larvae, the liver phenotypic changes were observed by stereomicroscopic imaging, acridine orange staining was used to detect hepatocyte apoptosis, histopathological of liver tissue vacuolization degree was analyzed, the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured. Transcriptome sequencing technology was further used to screen differentially expressed genes, and the expression levels of key genes were verified by qRT-PCR. Results The liver morphology of larvae zebrafish was abnormal after exposure to sub-lethal doses of strychnine. The pathological sections showed hepatocyte void formation, and the activities of ALT and AST were significantly increased (P < 0.05, 0.01). Transcriptomic analysis revealed significant enrichment of lipid metabolism pathway. Among them, key regulatory genes such as cell death-inducing DFF45-like effector B (CIDEB), cytochrome P450 family 7 subfamily A member 1 (CYP7A1), and fibroblast growth factor 19 (FGF19) showed abnormal expression. Conclusion Strychnine has obvious hepatotoxicity, and the mechanism of its hepatotoxicity may involve the abnormal expression of key genes regulating lipid metabolism pathway (such as CIDEB, CYP7A1, FGF19), and then lead to lipid metabolism disorders. This study provides theoretical reference for the safe and rational clinical use of Strychnos nux-vomica and related drug preparations containing strychnine.

R285.5

国家中医药管理局课题（GZY-KJS-2024-014）