摘要
目的利用转录组学探究对羟基苯甲醇对斑马鱼胚胎生长发育的影响及潜在作用机制。方法将6 hpf的斑马鱼胚胎在不同质量浓度的对羟基苯甲醇(0~0.5 mg·mL^(-1))中暴露120 h,分别记录胚胎的死亡率、孵化率、畸形率(包括心包水肿和卵黄囊水肿)、体长和心率;用吖啶橙染色观察仔鱼细胞凋亡情况;油红O染色观察斑马鱼仔鱼脂质积累情况;进一步利用转录组学技术测序,筛选差异表达基因并进行信号通路富集分析。结果与对照组比较,对羟基苯甲醇导致胚胎死亡率增加,心率下降,孵化率下降(P均<0.05);畸形率(心包水肿率、卵黄囊水肿率)增加,体长有明显缩短的趋势且呈浓度依赖性(P均<0.05);对羟基苯甲醇诱导仔鱼心脏细胞凋亡,增加斑马鱼卵黄中的脂质积累,使血管间隙中染色颜色较浅,造成了脂质代谢异常;转录组学结果表明,对羟基苯甲醇暴露导致的差异表达基因有5434个,KEGG显著富集到的信号通路包括嘌呤代谢、FOXO信号通路、嘧啶代谢、光传导、核苷酸代谢、范科尼贫血途径、药物代谢-其他酶等。结论对羟基苯甲醇暴露会引起斑马鱼胚胎生长发育异常,并且可能与抑制光传导和代谢相关通路,以及抑制ugt1b5、ugt5b4、ugt5b2、ugt1a1、opn1mw1、opn1mw2、gnb1a、gnat1基因的表达有关。
Objective To determine the effect of p-hydroxybenzyl alcohol on the embryonic development in zebrafish and its underlying mechanism by transcriptomics.Methods Zebrafish embryos at 6 hpf were exposed to varying concentrations of p-hydroxybenzyl alcohol(0~0.5 mg·mL^(-1))for 120 h.The embryo mortality,hatching rate,malformation rates(including pericardial edema and yolk sac edema),body length,and heart rate were recorded.Acridine orange staining was used to observe the larval apoptosis,while oil red O staining assessed the lipid accumulation;transcriptomic sequencing was conducted to identify differentially expressed genes for signaling pathway enrichment analysis.Results p-hydroxybenzyl alcohol increased the embryo mortality and the heart rate decreased,the hatching rate,with significant difference(all P<0.05).The rates of pericardial edema and yolk sac edema increased markedly,and a notable decrease in body length was found in a concentration-dependent manner(all P<0.05).More over,p-hydroxybenzyl alcohol induced apoptosis in larval heart cells;increased lipid accumulation in zebrafish yolk,and lighter vascular space staining,indicated disrupted lipid metabolism;transcriptomic analysis revealed 5434 differentially expressed genes after p-hydroxybenzyl alcohol exposure,and the most significantly differentially expressed pathways included normal substance metabolism-related pathways led by nucleic acid metabolism and phototransduction pathway.Conclusion p-hydroxybenzyl alcohol exposure may induce developmental abnormalities in zebrafish embryos,which is potentially associated with the inhibition of phototransduction/metabolic pathways and suppression of ugt1b5,ugt5b4,ugt5b2,ugt1a1,opn1mw1,opn1mw2,gnb1a,and gnat1.
作者
朱澳华
王平
夏婧
刘勇杰
昝俊峰
刘军锋
ZHU Ao-hua;WANG Ping;XIA Jing;LIU Yong-jie;ZAN Jun-feng;LIU Jun-feng(School of Pharmacy,Hubei University of Traditional Chinese Medicine,Wuhan 430065;Institute of Gerontology,Hubei University of Chinese Medicine,Wuhan 430065;Key Laboratory of TCM Resources and Compounds of Education Commission,Hubei University of Chinese Medicine,Wuhan 430065;Hubei Shizhen Laboratory,Wuhan 430065)
出处
《中南药学》
2025年第6期1704-1710,共7页
Central South Pharmacy
基金
湖北省自然科学基金-中医药创新发展联合基金资助项目(No.2024AFD285)。
关键词
对羟基苯甲醇
斑马鱼胚胎
生长发育
转录组学
p-hydroxybenzyl alcohol
zebrafish embryo
development and growth
transcriptomics
