两种有机磷酸酯对斑马鱼内分泌性别依赖性影响

苏可欣; 卢婷; 王鑫伟; 李梅

PDF(2642 KB)

中国环境科学 ›› 2026, Vol. 46 ›› Issue (2) : 1015-1027.

环境毒理与健康

两种有机磷酸酯对斑马鱼内分泌性别依赖性影响

苏可欣, 卢婷, 王鑫伟, 李梅

作者信息 +

Effects of two types of organophosphates on endocrine sex-dependent development in zebrafish

SU Ke-xin, LU Ting, WANG Xin-wei, LI Mei

Author information +

文章历史 +

摘要

为探讨有机磷酸酯阻燃剂磷酸三(1-氯-2-丙基)酯(TCPP)和间苯二酚双(二苯基)磷酸酯(RDP)对水生生物的内分泌干扰机制及性别差异化响应,以斑马鱼成鱼为模式生物,开展浓度分别为96h-LC₅₀的1%(400μg/L)和1‰(40 μg/L)的暴露实验.结果表明: TCPP和RDP暴露对斑马鱼雌/雄个体的肥满度、脑体指数和性腺体指数均未引发显著变化,但乙酰胆碱酯酶活性在所有暴露组均显著下降(7.80%~31.31%, P<0.01),其中雌鱼对TCPP的响应更为敏感.同时,各暴露组斑马鱼丙二醛水平升高(34.37%~166.25%),谷胱甘肽含量下降(29.51%~58.64%),显示TCPP和RDP均诱导了氧化应激反应.进一步研究发现,TCPP和RDP暴露显著扰动斑马鱼下丘脑-垂体-甲状腺轴(HPT)功能,导致甲状腺激素三碘甲状腺原氨酸水平下降、甲状腺素水平升高,且雌鱼受影响更为显著(P<0.01, P<0.05).基因表达分析显示,TCPP和RDP显著抑制雌鱼体内HPT轴相关基因crh和tshβ的表达(P<0.05),而雄鱼相关基因未见明显变化,进一步证实两种OPEs对HPT轴的干扰存在性别依赖性.此外,TCPP和RDP暴露干扰雌/雄斑马鱼性激素水平,其中雄鱼体内睾酮(T)水平显著升高,雌二醇(E2)水平显著降低(P<0.001),同时下丘脑-垂体-性腺轴(HPG)关键基因表达也受到影响,其中cyp19ala下调,抑制T向E2的转化,导致E2水平降低,影响雄鱼的精子生成及运动能力.综上,TCPP和RDP暴露均可通过HPT轴和HPG轴引发显著的内分泌干扰,且干扰具有明显的性别依赖性.本研究强调在环境污染物生态毒性评价中系统考虑性别差异的重要性.

Abstract

To elucidate the endocrine-disrupting mechanisms and sex-dependent responses of organophosphate flame retardants (OPEs), tris(1-chloro-2-propyl) phosphate (TCPP) and resorcinol bis(diphenyl) phosphate (RDP), adult zebrafish (Danio rerio) were employed as a model organism. Exposure experiments were conducted at two concentrations corresponding to 1% (400 μg/L) and 1‰ (40 μg/L) of the 96h median lethal concentration (LC₅₀). The results showed that neither TCPP nor RDP exposure caused significant changes in body condition factor, brain somatic index (BSI), or gonad somatic index (GSI) in either female or male zebrafish. However, acetylcholinesterase (AChE) activity was significantly inhibited in all exposed groups by 7.80%~31.31% (P<0.01), with females exhibiting greater sensitivity to TCPP. Meanwhile, malondialdehyde (MDA) levels were significantly elevated (34.37%~166.25%), whereas glutathione (GSH) contents were markedly reduced (29.51%~58.64%) indicating that both TCPP and RDP induce oxidative stress in zebrafish. Exposure to TCPP and RDP significantly disrupted the function of the hypothalamic-pituitary-thyroid (HPT) axis, as evidenced by decreasing triiodothyronine (T3) levels and increasing thyroxine (T4) levels, with more pronounced effects observed in females (P<0.01, P<0.05). Gene expression analysis further revealed that TCPP and RDP significantly downregulated the HPT axis-related genes crh and tshβ in females (P<0.05), whereas no significant alterations were detected in males, confirming a clear sex-dependent disruption of the HPT axis by these OPEs. In addition, TCPP and RDP exposure disturbed sex hormone homeostasis in zebrafish. In males, testosterone levels were significantly increased, while estradiol levels were markedly decreased (P<0.001). Consistent with these hormonal alterations, the expression of cyp19a1a, a key gene involved in the hypothalamic-pituitary-gonadal (HPG) axis, was significantly downregulated, inhibiting the conversion of testosterone to estradiol. This disruption may impair spermatogenesis and sperm motility in males zebrafish. Overall, TCPP and RDP exposure induced pronounced endocrine disruption via interfering with both the HPT and HPG axes, with distinct sex-dependent effects. This study underscores the importance of considering sex differences in the toxicological evaluation.

导出引用

苏可欣, 卢婷, 王鑫伟, 李梅. 两种有机磷酸酯对斑马鱼内分泌性别依赖性影响[J]. 中国环境科学. 2026, 46(2): 1015-1027

SU Ke-xin, LU Ting, WANG Xin-wei, LI Mei. Effects of two types of organophosphates on endocrine sex-dependent development in zebrafish[J]. China Environmental Science. 2026, 46(2): 1015-1027

中图分类号： X503

参考文献

[1] Xia M, Wang X, Xu J, et al. Tris (1-chloro-2-propyl) phosphate exposure to zebrafish causes neurodevelopmental toxicity and abnormal locomotor behavior [J]. Science of the Total Environment, 2021,758:143694.
[2] Castro-jimenez J, Gonzalez-gaya B, Pizarro M, et al. Organophosphate Ester flame retardants and plasticizers in the global oceanic atmosphere [J]. Environmental Science & Technology, 2016,50(23): 12831-12839.
[3] Chen M H, Ma W L. A review on the occurrence of organophosphate flame retardants in the aquatic environment in China and implications for risk assessment [J]. Science of the Total Environment, 2021,783: 147064.
[4] Wang Y, Sun H, Zhu H, et al. Occurrence and distribution of organophosphate flame retardants (OPFRs) in soil and outdoor settled dust from a multi-waste recycling area in China [J]. Science of the Total Environment, 2018,625:1056-1064.
[5] Wang Q, Zhao H, Bekele T G, et al. Organophosphate esters (OPEs) in wetland soil and Suaeda salsa from intertidal Laizhou Bay, North China: Levels, distribution, and soil-plant transfer model [J]. Science of the Total Environment, 2021,764:142891.
[6] Arukwe A, Carteny C C, Eggen T, et al. Novel aspects of uptake patterns, metabolite formation and toxicological responses in Salmon exposed to the organophosphate esters—Tris(2-butoxyethyl)- and tris(2-chloroethyl) phosphate [J]. Aquatic Toxicology, 2018,196: 146-153.
[7] Liu C, Zhang X, Deng J, et al. Effects of prochloraz or propylthiouracil on the cross-talk between the HPG, HPA, and HPT axes in zebrafish [J]. Environmental Science & Technology, 2011, 45(2):769-775.
[8] Hershman J M, Beck-peccoz P. Discoveries around the hypothalamic-pituitary-thyroid axis [J]. Thyroid: Official Journal of the American Thyroid Association, 2023,33(7):785-790.
[9] Kaprara A, Huhtaniemi I T. The hypothalamus-pituitary-gonad axis: Tales of mice and men [J]. Metabolism: clinical and experimental, 2018,86:3-17.
[10] Neumann A M, Schmidt C X, Brockmann R M, et al. Circadian regulation of endocrine systems [J]. Autonomic Neuroscience: Basic & Clinical, 2019,216:1-8.
[11] Hu W, Peng G, Lei W, et al. Endocrine disrupting toxicity of aryl organophosphate esters and mode of action [J]. Critical Reviews in Environmental Science & Technology, 2023,53(1):1-18.
[12] Mcgee S P, Cooper E M, Stapleton H M, et al. Early zebrafish embryogenesis is susceptible to developmental TDCPP exposure [J]. Environmental Health Perspectives, 2012,120(11):1585-1591.
[13] Yuan L, Li J, Zha J, et al. Targeting neurotrophic factors and their receptors, but not cholinesterase or neurotransmitter, in the neurotoxicity of TDCPP in Chinese rare minnow adults (Gobiocypris rarus) [J]. Environmental Pollution, 2016,208:670-677.
[14] Marklund A, Andersson B, Haglund P. Screening of organophosphorus compounds and their distribution in various indoor environments [J]. Chemosphere, 2003,53(9):1137-1146.
[15] Lee S, Jeong W, Kannan K, et al. Occurrence and exposure assessment of organophosphate flame retardants (OPFRs) through the consumption of drinking water in Korea [J]. Water Research, 2016,103: 182-188.
[16] Comfort N, Re D B. Sex-specific neurotoxic effects of organophosphate pesticides across the life course [J]. Current Environmental Health Reports, 2017,4(4):392-404.
[17] Wang X, Lu T, Yang B, et al. Exposure to resorcinol bis (diphenyl phosphate) induces colonization of alien microorganisms with potential impacts on the gut microbiota and metabolic disruption in male zebrafish [J]. Science of the Total Environment, 2024,932: 172892.
[18] Liu X, Lu X, Hong J, et al. Effects of long-term exposure to TDCPP in zebrafish (Danio rerio) - Alternations of hormone balance and gene transcriptions along hypothalamus-pituitary axes [J]. Animal Models and Experimental Medicine, 2022,5(3):239-247.
[19] Tan T, Yu R M K, Wu R S S, et al. Overexpression and knockdown of hypoxia-inducible factor 1 disrupt the expression of steroidogenic enzyme genes and early embryonic development in zebrafish [J]. Gene Regulation and Systems Biology, 2017,11:1177625017713193.
[20] Crump D, Chiu S, Kennedy S W. Effects of tris(1,3-dichloro-2-propyl) phosphate and tris(1-chloropropyl) phosphate on cytotoxicity and mRNA expression in primary cultures of avian hepatocytes and neuronal cells [J]. Toxicological Sciences: An Official Journal of the Society of Toxicology, 2012,126(1):140-148.
[21] Ihn Y, Cho Y, Lee Y, et al. Thyroid and sex hormone disrupting effects of DEHTP at different life stages of zebrafish (Danio rerio) [J]. Chemosphere, 2024,358:142105.
[22] Bianco A C, Dumitrescu A, Gereben B, et al. Paradigms of dynamic control of thyroid hormone signaling [J]. Endocrine reviews, 2019, 40(4):1000-1047.
[23] Tian D, Yu Y, Yu Y, et al. Tris(2-chloroethyl) phosphate exerts hepatotoxic impacts on zebrafish by disrupting hypothalamic- pituitary-thyroid and gut-liver axes [J]. Environmental Science & Technology, 2023,57(24):9043-9054.
[24] Yu L, Deng J, Shi X, et al. Exposure to DE-71 alters thyroid hormone levels and gene transcription in the hypothalamic-pituitary-thyroid axis of zebrafish larvae [J]. Aquatic Toxicology, 2010,97(3):226-233.
[25] Freitas J, Cano P, Craig-Vetic C, et al. Detection of thyroid hormone receptor disruptors by a novel stable in vitro reporter gene assay [J]. Toxicology in Vitro, 2011,25(1):257-266.
[26] Vujovic F, Farahani R M. Thyroid hormones and brain development: a focus on the role of mitochondria as regulators of developmental time [J]. Cells, 2025,14(3):150.
[27] Zhang L, Liu N, Shao J, et al. Bidirectional control of parathyroid hormone and bone mass by subfornical organ [J]. Neuron, 2023, 111(12):1914-1932.e6.
[28] Zha W, Hu W, Ge C, et al. Zebrafish as a model system for studying reproductive diseases [J]. Frontiers in Cell and Developmental Biology, 2024,12:1481634.
[29] Santhi J J, Issac P K, Velayutham M, et al. Reproductive toxicity of perfluorobutane sulfonate in zebrafish (Danio rerio): Impacts on oxidative stress, hormone disruption and HPGL axis dysregulation [J]. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 2025,289:110122.
[30] Santhakumar K. Neuroendocrine regulation of reproduction in fish - Mini review [J]. Aquaculture and Fisheries, 2024,9(3):437-446.
[31] Shi X, Wu R, Wang X, et al. Effects of 2,2¢,4,4¢-tetrabromodiphenyl ether (BDE-47) on reproductive and endocrine function in female zebrafish (Danio rerio) [J]. Ecotoxicology and Environmental Safety, 2022,248:114326.
[32] Meng X, Su S, Wei X, et al. Exposure to bisphenol A alternatives bisphenol AF and fluorene-9-bisphenol induces gonadal injuries in male zebrafish [J]. Ecotoxicology and Environmental Safety, 2023, 253:114634.
[33] Walczak-nowicka Ł J, Herbet M. Acetylcholinesterase inhibitors in the treatment of neurodegenerative diseases and the role of acetylcholinesterase in their pathogenesis [J]. International Journal of Molecular Sciences, 2021,22(17):9290.
[34] Salim S. Oxidative stress and the central nervous system [J]. The journal of pharmacology and experimental therapeutics, 2017,360(1): 201-205.
[35] Rodriguez P, Lopez-landa A, Romo-parra H, et al. Unraveling the ozone impact and oxidative stress on the nervous system [J]. Toxicology, 2024,509:153973.
[36] Li W, Zhu L, Du Z, et al. Acute toxicity, oxidative stress and DNA damage of three task-specific ionic liquids ([C₂NH₂MIm] BF₄, [MOEMIm] BF₄, and [HOEMIm] BF₄) to zebrafish (Danio rerio) [J]. Chemosphere, 2020,249:126119.
[37] Cao Z, Zou L, Wang H, et al. Exposure to diclofop-methyl induces immunotoxicity and behavioral abnormalities in zebrafish embryos [J]. Aquatic Toxicology, 2019,214:105253.
[38] Monteiro D A, Almeida J A de, Rantin F T, et al. Oxidative stress biomarkers in the freshwater characid fish, Brycon cephalus, exposed to organophosphorus insecticide Folisuper 600(methyl parathion) [J]. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 2006,143(2):141-149.
[39] Jiang N, Song P, Li X, et al. Dibutyl phthalate induced oxidative stress and genotoxicity on adult zebrafish (Danio rerio) brain [J]. Journal of Hazardous Materials, 2022,424:127749.
[40] Varticovski L, Stavreva D A, Mcgowan A, et al. Endocrine disruptors of sex hormone activities [J]. Molecular and Cellular Endocrinology, 2022,539:111415.
[41] Hoshi S, Saito N, Kusanagi K, et al. Adjuvant effects of fluoride on oral immunization of chickens [J]. Veterinary immunology and immunopathology, 1998,63(3):253-263.
[42] Shi X, Du Y, Lam P K S, et al. Developmental toxicity and alteration of gene expression in zebrafish embryos exposed to PFOS [J]. Toxicology and Applied Pharmacology, 2008,230(1):23-32.
[43] Spaan K, Haigis A C, Weiss J, et al. Effects of 25thyroid hormone disruptors on zebrafish embryos: A literature review of potential biomarkers [J]. Science of the Total Environment, 2019,656:1238- 1249.
[44] Baumann L, Holbech H, Schmidt-posthaus H, et al. Does hepatotoxicity interfere with endocrine activity in zebrafish (Danio rerio)? [J]. Chemosphere, 2020,238:124589.
[45] Wu K, Yue Y, Zhou L, et al. Disrupting Amh and androgen signaling reveals their distinct roles in zebrafish gonadal differentiation and gametogenesis [J]. Communications Biology, 2025,8(1):371.
[46] 钟圳,陈肇文,王有基,等.微纳米塑料和有机磷阻燃剂的联合毒性效应研究进展[J].生态毒理学报, 2022,17(6):37-68. Zhong Z, Chen Z, Wang Y J, et al. Research progress on the combined toxic effects of micro- and nanoplastics and organophosphorus flame retardants [J]. Asian Journal of Ecotoxicology, 2022,17(6):37-68.
[47] Wang Q, Lam J C W, Man Y C, et al. Bioconcentration, metabolism and neurotoxicity of the organophorous flame retardant 1,3-dichloro 2-propyl phosphate (TDCPP) to zebrafish [J]. Aquatic Toxicology, 2015,158:108-115.
[48] Liu X, Ji K, Choi K. Endocrine disruption potentials of organophosphate flame retardants and related mechanisms in H295R and MVLN cell lines and in zebrafish [J]. Aquatic Toxicology, 2012, 114-115:173-181.
[49] Freitas J S, Kupsco A, Diamante G, et al. Influence of temperature on the thyroidogenic effects of diuron and its metabolite 3,4-DCA in tadpoles of the American bullfrog (Lithobates catesbeianus) [J]. Environmental Science & Technology, 2016,50(23):13095-13104.
[50] 姜锦林,刘仁彬,张宇峰,等.壬基酚聚氧乙烯醚对成年雄性斑马鱼HPG轴相关基因表达的影响[J].中国环境科学, 2018,38(8): 3135-3142. Jiang J L, Liu R B, Zhang Y F, et al. Effects of nonylphenol ethoxylate on gene expression related to the hypothalamic-pituitary gonadal axis in adult male zebrafish [J]. China Environmental Science, 2018,38(8): 3135-3142.
[51] Zhang W, Sheng N, Wang M, et al. Zebrafish reproductive toxicity induced by chronic perfluorononanoate exposure [J]. Aquatic Toxicology (Amsterdam, Netherlands), 2016,175:269-276.
[52] Halliwell B, Gutteridge J M C. Free radicals in biology and medicine [M]. Oxford University Press, 2015:607-672.
[53] Niu L, Zhang Z, Zhang K, et al. Thyroid disruption and the association with multi-toxicity endpoints in zebrafish embryos exposed to hymexazol [J]. Ecotoxicology and Environmental Safety, 2025,302: 118697.
[54] Gaillard R C. Interaction between the hypothalamo-pituitary-adrenal axis and the immunological system [J]. Annales d’endocrinologie, 2001,62(2):155-163.
[55] Huang Y, Cai H, Han Y, et al. Mechanisms of heat stress on neuroendocrine and organ damage and nutritional measures of prevention and treatment in poultry [J]. Biology, 2024,13(11):926.
[56] Lombó M, Herráez P. The effects of endocrine disruptors on the male germline: an intergenerational health risk [J]. Biological reviews of the Cambridge Philosophical Society, 2021,96(4):1243-1262.
[57] Mancini A, Di Segni C, Raimondo S, et al. Thyroid hormones, oxidative stress, and inflammation [J]. Mediators of inflammation, 2016,2016:6757154.
[58] Behl C, Moosmann B. Antioxidant neuroprotection in Alzheimer’s disease as preventive and therapeutic approach [J]. Free Radical Biology and Medicine, 2002,33(2):182-191.