有机磷阻燃剂TCIPP和TnBP对斑马鱼胚胎发育影响的研究

Developmental effects of TCIPP and TnBP on zebrafish (Danio rerio) embryos

目的探讨磷酸三(2-氯丙基)酯(TCIPP)和磷酸三正丁酯(TnBP)对斑马鱼胚胎的生长发育毒性及潜在分子机制。方法以斑马鱼为模型,采用半静态法,将受精后2 h(hpf)斑马鱼胚胎暴露于TCIPP和TnBP(0.1、1、10、100、500和1000μmol/L)水溶液120 h,测定其生存率和孵化率,以及环境相关浓度(0.1和1μmol/L)下120 hpf斑马鱼转录组的变化。结果TCIPP和TnBP对斑马鱼胚胎的半数致死浓度分别是155.30和27.62μmol/L(96 hpf)、156.50和26.05μmol/L(120 hpf)。暴露于100μmol/L的TCIPP和10μmol/L的TnBP后斑马鱼72 hpf孵化率分别为(23.33±7.72)%和(91.67±2.97)%,与对照组相比均降低,差异有统计学意义(均P<0.05)。转录组结果显示,TnBP暴露导致的差异基因数多于TCIPP,并呈现“剂量-效应”关系。IPA通路富集分析提示共富集32条通路,涉及氧化应激、能量代谢、脂质代谢和核受体相关通路等。TCIPP和TnBP共同富集到3条通路,分别为甲状腺激素受体/视黄酮X受体和组成型雄烷受体/视黄酮X受体通路等。TnBP单独富集到肝脏X受体/视黄酮X受体和氧化应激相关通路等。结论TCIPP和TnBP对斑马鱼均具有生长发育毒性,TnBP毒性效应更强,TCIPP和TnBP诱导斑马鱼生长发育毒性的作用机制有所不同。

Objective To investigate the toxicity of tris(2-chloropropyl)phosphate(TCIPP)and tributyl phosphate(TnBP)on the growth and development of zebrafish embryos,as well as to explore the underlying mechanisms at the transcriptional level.Methods With zebrafish as a model,two hpf zebrafish embryos were exposed to TCIPP and TnBP(0.1,1,10,100,500,and 1000μmol/L)using the semi-static method,and their rates of lethality and hatchability were determined.The transcriptome changes of 120 hpf juvenile zebrafish exposed to environmentally relevant concentrations of 0.1 and 1μmol/L were measured.Results The 50%lethal concentrations(LC50)of TCIPP and TnBP for zebrafish embryos were 155.30 and 27.62μmol/L(96 hpf),156.5 and 26.05μmol/L(120 hpf),respectively.The 72 hpf hatching rates of TCIPP(100μmol/L)and TnBP(10μmol/L)were(23.33±7.72)%and(91.67±2.97)%,which were significantly decreased compared with the control group(P<0.05).Transcriptome analysis showed that TnBP had more differential genes(DEGs)than TCIPP,with a dose-response relationship.These DEGs were enriched in 32 pathways in total,including those involved in oxidative stress,energy metabolism,lipid metabolism,and nuclear receptor-related pathways,using the IPA pathway analysis.Among them,three enriched pathways overlapped between TCIPP and TnBP,including TR/RXR activation and CAR/RXR activation.Additionally,DEGs were also mapped onto pathways of LXR/RXR activation and oxidative stress for TnBP exposure only.Conclusion Both TCIPP and TnBP have growth and developmental toxicities in zebrafish embryos,with distinct biomolecular mechanisms,and TnBP has a stronger effect than TCIPP.