抗病毒药物依法韦仑对斑马鱼胚胎毒性机制的转录组分析  

Transcriptomic Analysis of the Toxicity Mechanism of the Antiviral Drug Contaminant Efavirenz to Zebrafish Embryos

作　　者：刘晨 李博洋 黎明瑞 徐佳芸 闫振广[1] LIU Chen;LI Boyang;LI Mingrui;XU Jiayun;YAN Zhenguang(State Key Laboratory of Environmental Criteria and Risk Assessment,Chinese Research Academy of Environmental Sciences,Beijing 100012,China;Ningbo Clinical Pathology Diagnostic Center,Ningbo 315021,China)

机构地区：[1]中国环境科学研究院,环境基准标准与风险管控全国重点实验室,北京100012 [2]宁波市临床病理诊断中心,浙江宁波315021

出　　处：《环境科学研究》2025年第2期437-446,共10页Research of Environmental Sciences

基　　金：国家自然科学基金面上项目(No.42277271)。

摘　　要：依法韦仑是一种被广泛使用的非核苷逆转录酶抑制物类(non-nucleoside reverse transcriptase inhibitor,NNRTI)抗病毒药物,在水环境中被广泛检出。目前抗病毒药物的毒理学研究主要集中于临床和哺乳动物模型等,然而对水生生物的毒性效应与致毒机制的研究也十分重要。本研究以斑马鱼为模式生物,通过胚胎急性暴露实验(以存活率、心率、体长为指标)、RNA提取转录组高通量测序、生物信息学分析及实时荧光定量PCR(qPCR)等方法探究典型抗病毒药物依法韦仑对斑马鱼胚胎的毒性作用机制,结果表明:(1)依法韦仑导致斑马鱼幼鱼存活率、心率、体长均出现剂量依赖性下降。(2)1 mg/L依法韦仑暴露96 h导致斑马鱼胚胎中1371个基因出现差异表达。(3)转录组分析表明,依法韦仑暴露导致的差异表达基因主要集中在141个信号通路上,其中差异表达最显著的通路包括异物代谢-细胞色素P450通路、以脂质代谢为首的正常物质代谢相关通路以及视觉发育相关通路(视黄醇代谢、光传导)。(4)通过qPCR验证发现,物质代谢相关的多条通路以及视黄醇代谢和光传导中的部分基因表达出现了下调,尤其是cyp3a65和cyp3c3基因表达下调最为显著。研究显示,依法韦仑可能的分子毒性机制是通过引起细胞色素P450家族失调,影响脂质、糖、氨基酸等物质代谢,并通过干扰视黄醇代谢、光传导等信号通路影响视觉发育,从而引起斑马鱼胚胎发育异常。Efavirenz,a widely used antiviral drug in the non-nucleoside reverse transcriptase inhibitor class,is frequently detected in aquatic environment.Currently,toxicological studies of antiviral drugs are mainly focused on clinical and mammalian models,and there are few studies on their toxic effects and virulence mechanisms of toxicity in aquatic organisms.In this study,zebrafish embryos were used as model organisms to investigate the toxic mechanisms of efavirenz through acute exposure experiments.Key metrics,including survival rate,heart rate,and body length,were assessed,and transcriptome analysis was performed using high-throughput RNA sequencing,bioinformatic analysis,and real-time fluorescence quantitative PCR(qPCR).The results showed that efavirenz stress resulted in a dose-dependent decrease in survival rate,heart rate,and body length in juvenile zebrafish.After 96 hours of exposure to 1 mg/L efavirenz,1371 genes were differentially expressed in zebrafish embryos.These genes were enriched in 141 signaling pathways,with significant differential expression in pathways related to xenobiotics metabolism by cytochrome P450,lipid metabolism,and visual development(retinol metabolism,phototransduction).Validation by qPCR confirmed downregulation of pathways related to material metabolism,especially retinol metabolism and phototransduction,with pronounced downregulation in genes such as cyp3a65 and cyp3c3.The findings showed that efavirenz disrupts lipids,glucose,and amino acids metabolism through cytochrome P450 dysregulation,and impairs visual development through interfering with retinol metabolism and phototransduction signaling pathways,leading to abnormal development of zebrafish embryos.

关 键 词：抗病毒药物 依法韦仑 斑马鱼 毒性机制 

分 类 号：X171.5[环境科学与工程—环境科学]