再生水景观利用对斑马鱼(Danio rerio)下丘脑-垂体-性腺(HPG)轴相关基因转录水平的影响
Effects of Reclaimed Water in Landscape Utilization on Gene Transcription of Hypothalamus-Pituitary-Gonadal (HPG) Axis in Zebrafish (Danio rerio)
-
摘要: 再生水对水体生物具有潜在的危害。为了评估再生水对水体生物的早期分子毒理效应，采用荧光定量多聚酶链式反应(polymerase chain reaction, PCR)技术分析再生水对斑马鱼下丘脑-垂体-性腺(hypothalamus-pituitary-gonadal, HPG)轴相关基因转录水平的影响。再生水暴露后，斑马鱼体长、体质量、生长状况因子(condition factor, K)、脑指数(brain-somatic index, BSI)和性腺指数(gonadosomatic index, GSI)无显著改变。再生水暴露7 d引起雄性斑马鱼脑雄激素受体(androgen receptor, Ar)、雌激素受体1 (estrogen receptor 1, Esr1)、卵泡刺激素β多肽(follicle stimulating hormone beta polypeptide, Fshb)、黄体生成素β多肽(luteinizing hormone beta polypeptide, Lhb)、促性腺激素释放激素(gonadotropin-releasing hormone 2, Gnrh2)和促性腺激素释放激素(gonadotropin-releasing hormone 3, Gnrh3)基因的转录水平显著下调(P<0.05)，并引起雌性斑马鱼脑Gnrh3基因转录水平显著上调(P<0.05)。再生水暴露7 d导致类固醇激素合成通路下游基因如Ar、Esr1、卵巢芳香化酶(ovarian cytochrome P450 aromatase, cyp19ala)、3β-羟基类固醇脱氢酶(3β-hydroxysteroid dehydrogenase, hsd3b)、17β-羟化类固醇脱氢酶3(17β-hydroxysteroid dehydrogenase type 3, hsd17b3)和20β-羟化类固醇脱氢酶(20β-hydroxysteroid dehydrogenase, hsd20b)在雄性斑马鱼睾丸中表达水平显著减少(P<0.05)；相反，雌性斑马鱼卵巢Esr1和11β-羟化酶(11β-hydroxylase, cyp11b)基因转录水平显著增加(P<0.05)。再生水对斑马鱼暴露14 d后，雄性斑马鱼脑Esr1、Fshb和Lhb基因表达水平显著下调(P<0.05)，睾丸Ar、Esr1、类固醇源性急性调节蛋白(steroidogenic acute regulatory, star)、cyp19a1a、11β-羟化类固醇脱氢酶2(11β-hydroxysteroid dehydrogenase 2, hsd11b2)、hsd20b和hsd17b3基因转录水平也显著降低(P<0.05)。然而再生水暴露14 d显著上调了雌性斑马鱼卵巢Esr1、孕激素受体(progesterone receptor, pgr)、cyp19a1a、hsd3b、hsd17b3和star基因转录水平(P<0.05)，并且下调了脑Gnrh3和Fshb的转录水平(P<0.05)。综上所述，本研究结果表明再生水显著影响了斑马鱼HPG轴相关基因转录水平，具有性别和暴露时间差异性，为再生水对斑马鱼生殖系统的影响提供了新的见解。
-
关键词:
- 再生水 /
- 斑马鱼(Danio rerio) /
- HPG轴 /
- 内分泌干扰效应
Abstract: Reclaimed water (RW) can be potentially harmful to aquatic organisms. To evaluate early molecular responses of reclaimed water on aquatic organisms, this study investigated the impact of RW on gene transcription of the hypothalamus-pituitary-gonadal (HPG) axis of adult zebrafish (Danio rerio) using the fluorescence quantitative polymerase chain reaction (PCR). After exposure, the total body length, wet weight, condition factor (K), brain-somatic index (BSI) and gonadosomatic index (GSI) did not change significantly. In brain, RW exposure for 7 d caused significant down-regulation in the transcripts of androgen receptor (Ar), estrogen receptor 1 (Esr1), follicle stimulating hormone beta polypeptide (Fshb), luteinizing hormone beta polypeptide (Lhb), gonadotropin-releasing hormone 2 (Gnrh2) and gonadotropin-releasing hormone 3 (Gnrh3) genes in male (P<0.05) and up-regulation in the transcript of Gnrh3 gene in female zebrafish (P<0.05). After exposure to RW for 7 d, the downstream genes of steroidogenic pathway like Ar, Esr1, ovarian cytochrome P450 aromatase (cyp19ala), 3β-hydroxysteroid dehydrogenase (hsd3b), 17β-hydroxysteroid dehydrogenase type 3 (hsd17b3) and 20β-hydroxysteroid dehydrogenase (hsd20b) expression were significantly reduced in testis of male zebrafish (P<0.05); on the contrary, the transcript of Esr1 and 11β-hydroxylase (cyp11b) genes were enhanced in ovary of female zebrafish (P<0.05). After exposure to RW for 14 d, it was found that the expression of Esr1, Fshb and Lhb genes was significantly down-regulated in brain of male zebrafish (P<0.05), and the expression of Ar, Esr1, steroidogenic acute regulatory (star), cyp19a1a, 11β-hydroxysteroid dehydrogenase 2 (hsd11b2), hsd20b and hsd17b3 genes was also significantly reduced in testis of male zebrafish (P<0.05). While in females, RW exposure for 14 d up-regulated the expression of Esr1, progesterone receptor (pgr), cyp19a1a, hsd3b, hsd17b3 and star in ovary (P<0.05), and meanwhile Gnrh3 and Fshb were down-regulated in brain (P<0.05). Overall, these findings exhibited RW have significant effects on the transcript of genes of HPG axis, showing sex and exposure time specific effects, which provide new insights of RW effects on zebrafish reproductive system. -
-
Li Z, Xiang X, Li M, et al. Occurrence and risk assessment of pharmaceuticals and personal care products and endocrine disrupting chemicals in reclaimed water and receiving groundwater in China [J]. Ecotoxicology and Environmental Safety, 2015, 119: 74-80 Sun J, Ji X W, Zhang R, et al. Endocrine disrupting compounds reduction and water quality improvement in reclaimed municipal wastewater: A field-scale study along Jialu River in North China [J]. Chemosphere, 2016, 157: 232-240 Sun Y, Huang H, Sun Y, et al. Ecological risk of estrogenic endocrine disrupting chemicals in sewage plant effluent and reclaimed water [J]. Environmental Pollution, 2013, 180: 339-344 Zhang H F, Jia Y T, Tang Z H, et al. Screening of chemicals with binding activities of liver X receptors from reclaimed waters [J]. Science of the Total Environment, 2020, 713: 136570 Wintgens T, Salehi F, Hochstrat R, et al. Emerging contaminants and treatment options in water recycling for indirect potable use [J]. Water Science and Technology: A Journal of the International Association on Water Pollution Research, 2008, 57(1): 99-107 Yuan J, van Dyke M I, Huck P M. Identification of critical contaminants in wastewater effluent for managed aquifer recharge [J]. Chemosphere, 2017, 172: 294-301 Altenburger R, Scholze M, Busch W, et al. Mixture effects in samples of multiple contaminants: An inter-laboratory study with manifold bioassays [J]. Environment International, 2018, 114: 95-106 Välitalo P, Massei R, Heiskanen I, et al. Effect-based assessment of toxicity removal during wastewater treatment [J]. Water Research, 2017, 126: 153-163 Busch W, Schmidt S, Kühne R, et al. Micropollutants in European rivers: A mode of action survey to support the development of effect-based tools for water monitoring [J]. Environmental Toxicology and Chemistry, 2016, 35(8): 1887-1899 Ren X H, Kou Y Y, Kim T, et al. Toxicity study of reclaimed water on human embryonic kidney cells [J]. Chemosphere, 2017, 189: 390-398 Planelló R, Herrero O, García P, et al. Developmental/reproductive effects and gene expression variations in Chironomus riparius after exposure to reclaimed water and its fortification with carbamazepine and triclosan [J]. Water Research, 2020, 178: 115790 Aquilino M, Martínez-Guitarte J L, García P, et al. Combining the assessment of apical endpoints and gene expression in the freshwater snail Physa acuta after exposure to reclaimed water [J]. Science of the Total Environment, 2018, 642: 180-189 Huang W H, Ai W M, Lin W W, et al. Identification of receptors for eight endocrine disrupting chemicals and their underlying mechanisms using zebrafish as a model organism [J]. Ecotoxicology and Environmental Safety, 2020, 204: 111068 Huang Y, Wang X, Zhang J, et al. Impact of endocrine-disrupting chemicals on reproductive function in zebrafish (Danio rerio) [J]. Reproduction in Domestic Animals, 2015, 50(1): 1-6 Bassi I, ANDRé V, Marelli F, et al. The zebrafish: An emerging animal model for investigating the hypothalamic regulation of reproduction [J]. Minerva Endocrinologica, 2016, 41(2): 250-265 Ankley G T, Johnson R D. Small fish models for identifying and assessing the effects of endocrine-disrupting chemicals [J]. ILAR Journal, 2004, 45(4): 469-483 Maharajan K, Muthulakshmi S, Karthik C, et al. Pyriproxyfen induced impairment of reproductive endocrine homeostasis and gonadal histopathology in zebrafish (Danio rerio) by altered expression of hypothalamus-pituitary-gonadal (HPG) axis genes [J]. Science of the Total Environment, 2020, 735: 139496 Cao J L, Wang G D, Wang T Y, et al. Copper caused reproductive endocrine disruption in zebrafish (Danio rerio) [J]. Aquatic Toxicology, 2019, 211: 124-136 Liang Y Q, Huang G Y, Ying G G, et al. A time-course transcriptional kinetics of the hypothalamic-pituitary-gonadal and hypothalamic-pituitary-adrenal axes in zebrafish eleutheroembryos after exposure to norgestrel [J]. Environmental Toxicology and Chemistry, 2015, 34(1): 112-119 Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method [J]. Methods, 2001, 25(4): 402-408 Guo D M, Liu W P, Yao T S, et al. Combined endocrine disruptive toxicity of malathion and cypermethrin to gene transcription and hormones of the HPG axis of male zebrafish (Danio rerio) [J]. Chemosphere, 2021, 267: 128864 Muthulakshmi S, Hamideh P F, Habibi H R, et al. Mycotoxin zearalenone induced gonadal impairment and altered gene expression in the hypothalamic-pituitary-gonadal axis of adult female zebrafish (Danio rerio) [J]. Journal of Applied Toxicology, 2018, 38(11): 1388-1397 Crowder C M, Lassiter C S, Gorelick D A. Nuclear androgen receptor regulates testes organization and oocyte maturation in zebrafish [J]. Endocrinology, 2017, 159(2): 980-993 Qiu W H, Fang M J, Liu J Y, et al. In vivo actions of bisphenol F on the reproductive neuroendocrine system after long-term exposure in zebrafish [J]. Science of the Total Environment, 2019, 665: 995-1002 Ji K, Hong S, Kho Y, et al. Effects of bisphenol S exposure on endocrine functions and reproduction of zebrafish [J]. Environmental Science & Technology, 2013, 47(15): 8793-8800 Liu X S, Ji K, Jo A, et al. Effects of TDCPP or TPP on gene transcriptions and hormones of HPG axis, and their consequences on reproduction in adult zebrafish (Danio rerio) [J]. Aquatic Toxicology, 2013, 134-135: 104-111 靳亚茹, 刘红玲, 韩志华, 等. BDE-28及BDE-99对斑马鱼早期生命阶段HPT、HPG和HPA轴功能基因表达水平的影响[J]. 生态毒理学报, 2018, 13(1): 106-118 Jin Y R, Liu H L, Han Z H, et al. Effects of BDE-28 and BDE-99 on functional gene expression along HPT,HPG and HPA axes during early life stages of zebrafish [J]. Asian Journal of Ecotoxicology, 2018, 13(1): 106-118 (in Chinese)
Ji K, Liu X S, Lee S, et al. Effects of non-steroidal anti-inflammatory drugs on hormones and genes of the hypothalamic-pituitary-gonad axis, and reproduction of zebrafish [J]. Journal of Hazardous Materials, 2013, 254-255: 242-251 Xu Q L, Wu D, Dang Y, et al. Reproduction impairment and endocrine disruption in adult zebrafish (Danio rerio) after waterborne exposure to TBOEP [J]. Aquatic Toxicology, 2017, 182: 163-171 Teng M M, Qi S Z, Zhu W T, et al. Effects of the bioconcentration and parental transfer of environmentally relevant concentrations of difenoconazole on endocrine disruption in zebrafish (Danio rerio) [J]. Environmental Pollution, 2018, 233: 208-217 Sun W, Jia Y L, Ding X S, et al. Combined effects of pentachlorophenol and its byproduct hexachlorobenzene on endocrine and reproduction in zebrafish [J]. Chemosphere, 2019, 220: 216-226 Fernandes D, Schnell S, Porte C. Can pharmaceuticals interfere with the synthesis of active androgens in male fish? An in vitro study [J]. Marine Pollution Bulletin, 2011, 62(10): 2250-2253 Wang Y J, Ge W. Cloning of zebrafish ovarian carbonyl reductase-like 20β-hydroxysteroid dehydrogenase and characterization of its spatial and temporal expression [J]. General and Comparative Endocrinology, 2002, 127(3): 209-216 梁燕秋. 孕激素物质黄体酮和甲炔诺酮对斑马鱼不同发育阶段的内分泌干扰效应[D]. 北京: 中国科学院大学, 2016: 87-104 Liang Y Q. Endocrine disrupting effects of progestins (progesterone and norgestrel) on the different stages of zebrafish(Danio rerio) [D]. Beijing: University of Chinese Academy of Sciences, 2016: 87 -104 (in Chinese)
Yi X H, Liu H H, Lu Y T, et al. Altered serum levels of sex steroids and biotransformation enzyme activities by long-term alachlor exposure in crucian carp (Carassius auratus) [J]. Bulletin of Environmental Contamination and Toxicology, 2007, 79(3): 283-287 McKinlay R, Plant J A, Bell J N B, et al. Endocrine disrupting pesticides: Implications for risk assessment [J]. Environment International, 2008, 34(2): 168-183 Mylchreest E, Cattley R C, Foster P M D. Male reproductive tract malformations in rats following gestational and lactational exposure to di(n-butyl) phthalate: An antiandrogenic mechanism? [J]. Toxicological Sciences, 1998, 43(1): 47-60 Mylchreest E, Sar M, Cattley R C, et al. Disruption of androgen-regulated male reproductive development by di(n-butyl) phthalate during late gestation in rats is different from flutamide [J]. Toxicology and Applied Pharmacology, 1999, 156(2): 81-95 Kassotis C D, Tillitt D E, Davis J W, et al. Estrogen and androgen receptor activities of hydraulic fracturing chemicals and surface and ground water in a drilling-dense region [J]. Endocrinology, 2014, 155(3): 897-907 Folmar L C, Denslow N D, Rao V, et al. Vitellogenin induction and reduced serum testosterone concentrations in feral male carp (Cyprinus carpio) captured near a major metropolitan sewage treatment plant [J]. Environmental Health Perspectives, 1996, 104(10): 1096-1101 -
点击查看大图
计量
- 文章访问数: 1824
- HTML全文浏览数: 1824
- PDF下载数: 51
- 施引文献: 0
下载:
