雄激素群勃龙对斑马鱼幼鱼尾鳍再生的影响机制研究
Mechanisms of Effects of Androgen 17β-Trenbolone on Caudal Fin Regeneration in Zebrafish Larvae
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摘要: 生物体的组织损伤是无法避免的,保持一定的组织修复和再生能力对于生物体而言极为重要。研究发现,包括环境雌激素在内的许多环境污染物能够影响鱼类的组织修复与再生,但雄激素是否有类似的毒性效应则未见报道。本研究以群勃龙作为雄激素的代表,首先通过斑马鱼幼鱼尾鳍再生模型,确认群勃龙能否影响斑马鱼幼鱼尾鳍再生;进一步地,通过与雄激素受体拮抗剂氟他胺的联合暴露,检测了雄激素受体、免疫系统和再生信号通路相关基因的转录水平变化,以及炎症细胞的招募与迁移,揭示雄激素干扰组织修复与再生的可能机理。研究表明,群勃龙(1 000 ng·L-1)能抑制斑马鱼幼鱼尾鳍再生,而与氟他胺(30~300 μg·L-1)联合暴露之后,其抑制效应被显著缓解。在基因转录水平上,雄激素受体基因mRNA的变化确证了2个化合物分别对雄激素受体的激动或拮抗效应。同时,群勃龙单独暴露影响了免疫系统和再生信号通路相关基因的转录水平,而在联合暴露后,多数基因的转录水平恢复,并与对照组无差异;再生尾鳍处中性粒细胞数量变化也有类似的趋势。由此可以推断,群勃龙可能是通过雄激素受体途径,并随之干扰免疫系统和再生信号通路,从而抑制斑马鱼尾鳍再生。考虑到水体环境中雄激素的普遍存在,本研究为评价雄激素的生态风险提供了新的视角。Abstract: Tissue damages are unavoidable for multicellular organisms across the whole life span, and the ability of tissue repair and regeneration is critical for their fitness and survival in the wild. It has been documented that several pollutants, including environmental estrogens, can disrupt the regenerative capacity in fish. However, whether the exposure to androgens will result in similar effects remains unclear. In this study, in order to determine the effects of androgen on the regenerative capacity and reveal the underlying mechanism, a zebrafish larval fin regeneration model was employed, and the fish following fin amputation were exposed to an androgen receptor agonist, 17β-trenbolone (TREN) alone, and in combination with the androgen receptor antagonist, flutamide (FLUT). It was found that exposure to 17β-trenbolone (1 000 ng·L-1) caused the inhibition of caudal fin regeneration, which was ameliorated after combination with flutamide (30~300 μg·L-1). Consistent with expectation, the change of mRNA level for androgen receptor confirmed their modes of action as an agonist or antagonist. Moreover, the changed transcript level of genes related to regeneration or immune response caused by 17β-trenbolone were mostly counteracted by flutamide, and the similar changing pattern was found for the number of neutrophils in the regenerating fin. Therefore, all these observations demonstrated that 17β-trenbolone could inhibit the fin regeneration in zebrafish larvae, which possibly due to the activation of androgen receptor and the consequent changes in signaling networks regulating regeneration and the immune system. Given the occurrence of androgens in the aquatic environment, and the conservation of cellular and molecular mechanism in tissue regeneration across species, this study will be helpful for a comprehensive ecotoxicological risk assessment of androgens.
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Yoshinari N, Kawakami A. Mature and juvenile tissue models of regeneration in small fish species[J]. The Biological Bulletin, 2011, 221(1):62-78 Sehring I M, Jahn C, Weidinger G. Zebrafish fin and heart:What's special about regeneration?[J]. Current Opinion in Genetics & Development, 2016, 40:48-56 李礼, 罗凌飞. 以斑马鱼为模式动物研究器官的发育与再生[J]. 遗传, 2013, 35(4):321-332 Li L, Luo L F. Zebrafish as the model system to study organogenesis and regeneration[J]. Hereditas, 2013, 35(4):321-332 (in Chinese)
Sehring I M, Weidinger G. Recent advancements in understanding fin regeneration in zebrafish[J]. Wiley Interdisciplinary Reviews Developmental Biology, 2020, 9(1):e367 Mathew L K, Andreasen E A, Tanguay R L. Aryl hydrocarbon receptor activation inhibits regenerative growth[J]. Molecular Pharmacology, 2006, 69(1):257-265 Zodrow J M, Tanguay R L. 2,3,7,8-tetrachlorodibenzo-p-dioxin inhibits zebrafish caudal fin regeneration[J]. Toxicological Sciences, 2003, 76(1):151-161 Mathew L K, Sengupta S, Kawakami A, et al. Unraveling tissue regeneration pathways using chemical genetics[J]. The Journal of Biological Chemistry, 2007, 282(48):35202-35210 Sengupta S, Bisson W H, Mathew L K, et al. Alternate glucocorticoid receptor ligand binding structures influence outcomes in an in vivo tissue regeneration model[J]. Comparative Biochemistry and Physiology Toxicology & Pharmacology, 2012, 156(2):121-129 Hirose K, Payumo A Y, Cutie S, et al. Evidence for hormonal control of heart regenerative capacity during endothermy acquisition[J]. Science, 2019, 364(6436):184-188 Sun L W, Gu L Q, Tan H N, et al. Effects of 17α[WT《Times New Roman》]-ethinylestradiol on caudal fin regeneration in zebrafish larvae[J]. The Science of the Total Environment, 2019, 653:10-22 Parks L G, Lambright C S, Orlando E F, et al. Masculinization of female mosquitofish in Kraft mill effluent-contaminated Fenholloway River water is associated with androgen receptor agonist activity[J]. Toxicological Sciences:An Official Journal of the Society of Toxicology, 2001, 62(2):257-267 Ankley G T, Coady K K, Gross M, et al. A critical review of the environmental occurrence and potential effects in aquatic vertebrates of the potent androgen receptor agonist 17β[WT《Times New Roman》]-trenbolone[J]. Environmental Toxicology and Chemistry, 2018, 37(8):2064-2078 Bartelt-Hunt S L, Snow D D, Kranz W L, et al. Effect of growth promotants on the occurrence of endogenous and synthetic steroid hormones on feedlot soils and in runoff from beef cattle feeding operations[J]. Environmental Science & Technology, 2012, 46(3):1352-1360 Ankley G T, Bencic D C, Breen M S, et al. Endocrine disrupting chemicals in fish:Developing exposure indicators and predictive models of effects based on mechanism of action[J]. Aquatic Toxicology, 2009, 92(3):168-178 Sun L W, Peng T, Liu F, et al. Transcriptional responses in male Japanese medaka exposed to antiandrogens and antiandrogen/androgen mixtures[J]. Environmental Toxicology, 2016, 31(11):1591-1599 Gu L Q, Tian L, Gao G, et al. Inhibitory effects of polystyrene microplastics on caudal fin regeneration in zebrafish larvae[J]. Environmental Pollution, 2020, 266(Pt 3):114664 Renshaw S A, Loynes C A, Trushell D M I, et al. A transgenic zebrafish model of neutrophilic inflammation[J]. Blood, 2006, 108(13):3976-3978 Rehberger K, Werner I, Hitzfeld B, et al. 20 Years of fish immunotoxicology-What we know and where we are[J]. Critical Reviews in Toxicology, 2017, 47(6):509-535 Casanova-Nakayama A, Wenger M, Burki R, et al. Endocrine disrupting compounds:Can they target the immune system of fish?[J]. Marine Pollution Bulletin, 2011, 63(5-12):412-416 Milla S, Depiereux S, Kestemont P. The effects of estrogenic and androgenic endocrine disruptors on the immune system of fish:A review[J]. Ecotoxicology, 2011, 20(2):305-319 Ingerslev H C, Lunder T, Nielsen M E. Inflammatory and regenerative responses in salmonids following mechanical tissue damage and natural infection[J]. Fish & Shellfish Immunology, 2010, 29(3):440-450 Peiris T H, Hoyer K K, Oviedo N J. Innate immune system and tissue regeneration in planarians:An area ripe for exploration[J]. Seminars in Immunology, 2014, 26(4):295-302 Tal T L, Franzosa J A, Tanguay R L. Molecular signaling networks that choreograph epimorphic fin regeneration in zebrafish-A mini-review[J]. Gerontology, 2010, 56(2):231-240 Wehner D, Weidinger G. Signaling networks organizing regenerative growth of the zebrafish fin[J]. Trends in Genetics, 2015, 31(6):336-343 Wehner D, Cizelsky W, Vasudevaro M D, et al. Wnt/β[WT《Times New Roman》]-catenin signaling defines organizing centers that orchestrate growth and differentiation of the regenerating zebrafish caudal fin[J]. Cell Reports, 2014, 6(3):467-481 Shibata E, Yokota Y, Horita N, et al. FGF signalling controls diverse aspects of fin regeneration[J]. Development, 2016, 143(16):2920-2929 Massart S, Redivo B, Flamion E, et al. The trenbolone acetate affects the immune system in rainbow trout, Oncorhynchus mykiss[J]. Aquatic Toxicology, 2015, 163:109-120 Mescher A L, Neff A W, King M W. Inflammation and immunity in organ regeneration[J]. Developmental & Comparative Immunology, 2017, 66:98-110 Poss K D. Advances in understanding tissue regenerative capacity and mechanisms in animals[J]. Nature Reviews Genetics, 2010, 11(10):710-722 -
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