中国药物警戒 ›› 2026, Vol. 23 ›› Issue (4): 398-404.
DOI: 10.19803/j.1672-8629.20260018

• 基于斑马鱼模型的中药毒性特征研究专栏 • 上一篇    下一篇

基于CYP抑制剂和转录组学研究杠柳毒苷通过抑制HNF4α-CYP轴诱导斑马鱼肝损伤

刘雨欣1, 生玉涵1, 于蔚洁1, 程云哲1, 李嘉琪1,2, 孙燕1#, 赵崇军1,2,*   

  1. 1北京中医药大学中医学院,北京 102488;
    2北京中医药大学中药学院,中药品质评价北京重点实验室,国家中医药管理局中药炮制技术传承基地,北京 102488
  • 收稿日期:2026-01-07 出版日期:2026-04-15 发布日期:2026-04-15
  • 通讯作者: *赵崇军,男,博士,助理研究员,中药安全性评价与毒性物质基础筛选。E-mail: 1014256537@qq.com。#为共同通信作者。
  • 作者简介:刘雨欣,女,硕士,中医学。
  • 基金资助:
    国家自然科学基金资助项目（82204753）; 北京市自然科学基金资助项目(7252240); 国家中医药传承创新团队项目（ZYYCXTD-C-202401-3）

Periplocin-Induced Hepatotoxicity in Zebrafish via Inhibition of the HNF4α-CYP Axis:a Combination of CYP Inhibitors and Transcriptomics

LIU Yuxin1, SHENG Yuhan1, YU Weijie1, CHENG Yunzhe1, LI Jiaqi1,2, SUN Yan1#, ZHAO Chongjun1,2,*   

  1. 1School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 102488, China;
    2School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing Key Laboratory for Quality Evaluation of Chinese Materia Medica, Traditional Chinese Medicine Processing Technology Inheritance Base of National Administration of Traditional Chinese Medicine, Beijing 102488, China
  • Received:2026-01-07 Online:2026-04-15 Published:2026-04-15

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摘要： 目的 探讨杠柳毒苷诱导肝毒性的作用机制,为杠柳毒苷研究提供参考。方法 基于斑马鱼模型,将受精后第4天的斑马鱼幼鱼作为模型,联用细胞色素P450（Cytochrome P450, CYP）抑制剂,通过转基因斑马鱼肝脏荧光表型及苏木精-伊红染色确认杠柳毒苷的肝毒性特征;采用转录组测序技术和实时荧光定量PCR（Real-Time Quantitative PCR, RT-qPCR）对相关机制进行验证。结果 研究发现,杠柳毒苷暴露导致斑马鱼肝脏荧光强度呈剂量依赖性降低,并引起肝细胞空泡化、排列紊乱等病理表现;CYP抑制剂处理进一步加剧了上述毒性效应,提示杠柳毒苷的肝毒性与其代谢特征密切相关。转录组学分析显示,杠柳毒苷处理会引起肝脏基因转录谱的显著改变,主要表现为CYP外源物代谢通路、脂质代谢及胆汁酸合成通路受到显著抑制。RT-qPCR实验证实,关键转录因子hnf4a及其下游Ⅰ相代谢酶cyp3a65、cyp2p6,以及Ⅱ相代谢酶gstp1及脂质代谢调控基因pparab和胆汁酸调控基因cyp8b1的表达水平显著下调（P<0.05）。结论 杠柳毒苷可能通过抑制上游关键转录因子肝细胞核因子4α（Hepatocyte Nuclear Factor 4α, HNF4α）的表达,抑制下游CYP等解毒酶系统,并导致过氧化物酶体增殖物激活受体α（Peroxisome Proliferator-Activated Receptor α, PPARα）相关的脂质代谢通路受到转录抑制,破坏肝脏稳态,最终导致解毒能力丧失与代谢紊乱,从而诱发严重的肝损伤。

关键词: 杠柳毒苷, 肝毒性, 斑马鱼, 转录组学, 细胞色素P450

Abstract: Objective To elucidate the underlying mechanisms of periplocin-induced hepatotoxicity using a zebrafish model, combined with cytochrome P450 (CYP) inhibitor interventions and transcriptomics. Methods Zebrafish larvae at 4 days post-fertilization (4 dpf) were used as the experimental model. Hepatotoxicity was evaluated by analyzing liver phenotypes in transgenic zebrafish and performing hematoxylin-eosin (H&E) staining following combined exposure to periplocin and CYP inhibitors. Subsequently, transcriptome sequencing and real-time quantitative PCR (RT-qPCR) were employed to uncover and validate the molecular mechanisms. Results Periplocin exposure caused a dose-dependent decrease in liver fluorescence intensity and induced pathological damage, including hepatocyte vacuolization and disordered arrangement. Notably, co-treatment with CYP inhibitors exacerbated these toxic effects, suggesting that the hepatotoxicity of periplocin was closely associated with its metabolic clearance. Transcriptomic analysis revealed significant alterations in the hepatic gene transcription profile, characterized primarily by the suppression of pathways related to CYP-mediated xenobiotic metabolism, lipid metabolism, and bile acid synthesis. RT-qPCR confirmed the significant downregulation of the key transcription factor hnf4a and its downstream phaseⅠmetabolic enzymes (cyp3a65, cyp2p6), phaseⅡmetabolic enzyme (gstp1), as well as the lipid metabolism regulator pparab and the bile acid regulator cyp8b1 (P<0.05). Conclusion Periplocin can induce severe liver injury by suppressing the expression of the upstream core transcription factor-hepatocyte nuclear factor 4α (HNF4α). This suppression triggers a transcriptional inhibition cascade affecting downstream CYP detoxification systems and the PPARα lipid metabolism pathway, leading to disrupted hepatic homeostasis, diminished detoxification capacity, and metabolic disorders.

Key words: Periplocin, Hepatotoxicity, Zebrafish, Transcriptomics, Cytochrome P450

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