严骁2,
付惠玲3,
庄僖2,
郑晶2,,,
麦碧娴4,
杨中艺3,
于云江2
1. 中山大学农学院, 深圳 518107;
2. 生态环境部华南环境科学研究所, 国家环境保护环境污染健康风险评价重点实验室, 广州 510530;
3. 湖南工学院安全与环境工程学院, 衡阳 421002;
4. 中国科学院广州地球化学研究所, 有机地球化学国家重点实验室, 广州 510640
作者简介: 何春桃(1989-),女,副教授,研究方向为生态学,E-mail:hecht3@mail.sysu.edu.cn.
通讯作者: 郑晶,zhengjing@scies.org ;
基金项目: 国家自然科学基金青年基金资助项目（21707177）；广州市科技计划项目（201804010074）；国家自然科学基金重点项目（41931290）中图分类号: X171.5
TPhP Induced Lipid Metabolism Disruption Mediated by miRNA in Hepatocyte
He Chuntao1,2,Yan Xiao2,
Fu Huiling3,
Zhuang Xi2,
Zheng Jing2,,,
Mai Bixian4,
Yang3,
Yu Yunjiang2
1. School of Agriculture, Sun Yat-sen University, Shenzhen 518107, China;
2. State Environmental Protection Key Laboratory of Environmental Pollution Health Risk Assessment, South China Institute of Environmental Sciences, Ministry of Ecology and Environment, Guangzhou 510530, China;
3. School of Safety and Environmental Engineering, Hunan Institute of Technology, Hengyang 421002, China;
4. Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, State Key Laboratory of Organic Geochemistry, Guangzhou 510640, China
Corresponding author: Zheng Jing,zhengjing@scies.org ;
CLC number: X171.5
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摘要:磷酸三苯酯（triphenyl phosphate，TPhP）是近年来广泛应用于电子产品的磷系阻燃剂，其脂类代谢干扰作用受到广泛关注，而microRNAs（miRNAs）在TPhP脂类代谢干扰过程中的调控作用仍鲜有报道。本研究通过探讨TPhP暴露条件下人肝细胞的细胞活性、TPhP清除速率与miRNAs表达调控特征，并与已获得转录组数据联合分析，明确TPhP经miRNA介导的脂类代谢干扰作用。在不同浓度TPhP暴露处理48 h后，人肝细胞的细胞活性随着暴露浓度呈先上升后下降的趋势，半数致死效应剂量为46.7 μmol·L-1。在5 μg·mL-1的TPhP暴露3 h和48 h后，TPhP的清除率分别为73.9%和85.1%。人肝细胞多个miRNAs表达差异显著，差异表达miRNAs所调控的靶基因主要参与代谢、脂肪酸合成、类固醇合成及癌症相关通路。通过转录组关联分析，差异表达miR-34c-5p、miR-301a-5p和miR-7等多个miRNAs对脂类代谢相关通路的关键基因具有调控作用，并构成miRNA-mRNA调控网络。综上所述，人肝细胞对TPhP具有较高的清除效率，TPhP暴露诱导miRNA差异表达所介导的脂类代谢干扰作用是TPhP对人肝细胞的主要毒性作用，miR-7和miR-4484等miRNAs能作为TPhP胁迫下脂类代谢干扰作用的潜在生物指示物。
关键词: 磷酸三苯酯/
人肝细胞/
脂类代谢干扰/
miRNAs
Abstract:Triphenyl phosphate (TPhP) is one of the novel alternative retardants with increasing utility. As a potential obesogen, the direct disruption of TPhP on lipid metabolism in hepatocyte has been widely studied. However, the microRNAs (miRNAs) regulation patterns of lipid metabolism disruption effects in hepatocyte under TPhP exposure remain unknown. This study aimed to elucidate the adverse effect of TPhP on hepatocyte through cell viability, TPhP clearance, and epigenetic miRNA alterations of hepatocytes at short (3 h) and long term (48 h) exposures to TPhP. The cell viability of hepatocyte raised at the low TPhP concentration treatments but dramatically decreased after the 10 μg·mL-1TPhP treatment, and concentration for 50% of maximal inhibitory effect was 46.7 μmol·L-1. The clearance rates of TPhP in hepatocyte were 73.9% and 85.1% after 3 h and 48 h incubations, respectively. Plenty of miRNAs were differentially expressed under TPhP treatments of 5 μg·mL-1. The predicted target genes of these differential expressed miRNAs were mainly involved in metabolic, fatty acid biosynthesis, steroid biosynthesis and cancer related pathways. The regulation network of miRNA-mRNA was constructed according to the correlations between miR-34c-5p, miR-301a-5p, and miR-7 and lipid metabolism related genes. Lipid metabolism disruption by TPhP was achieved by a proposed integration network of differential expressed mRNAs and miRNAs. Our study provides important insight in potential miRNAs biomarkers such as miR-4484 and miR-7 and the miRNA-mediated lipid metabolism disruption effect on human hepatocytes under TPhP treatment.
Key words:triphenyl phosphate/
hepatocyte/
lipid disruption/
miRNAs.
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