郝小吉;尚小娜;于海华;么建萍;韩璐;刘晓晖;李亚晨;邵静;

• 1:大连医科大学公共卫生学院大连市血液病重点实验室辽宁省造血干细胞移植医学中心辽宁省造血干细胞移植临床转化医学研究重点实验室
• 2:康龙化成(北京)新药技术股份有限公司
• 3:大连市妇幼保健院

摘要(Abstract)：

六溴环十二烷(hexabromocyclododecane,HBCD)与多溴联苯醚(polybrominated diethyl ethers,PBDEs)复合污染体系,对人类健康尤其神经系统所造成的潜在危害及其机制一直是笔者课题组的研究方向。HBCD是广泛使用的溴化阻燃剂,与PBDEs一样,会通过干扰内分泌系统、影响甲状腺激素分泌及损伤神经系统,对生物体产生发育神经毒性。作为系列研究之一,本研究以H4人脑神经胶质瘤细胞和SK-N-AS人神经母细胞瘤细胞为体外生物模型,通过观察HBCD对H4细胞Ⅱ型脱碘酶(Dio2)和SK-N-AS细胞Ⅲ型脱碘酶(Dio3)表达的调控,初步探讨了HBCD对神经系统局部甲状腺激素水平的潜在影响。H4细胞和SK-N-AS细胞分别暴露于0、1、3和9μmol·L-1HBCD 24 h后,采用MTT法检测细胞活力,Western Blot和RT-PCR法分别分析Dio2和Dio3蛋白和基因的表达,酶联免疫吸附测定(ELISA)法检测H4细胞脑源性神经细胞营养因子(BDNF)的分泌。结果表明,HBCD以剂量依赖方式降低H4细胞和SK-N-AS细胞生存率,引起H4细胞Dio2蛋白和基因表达下调,而致SK-N-AS细胞Dio3蛋白和基因表达上调。此外,HBCD还降低H4细胞BDNF的分泌。这表明,HBCD很可能通过影响神经和胶质细胞脱碘酶的表达,影响脑局部甲状腺激素水平,从而引起神经系统损伤及发育神经毒性。

关键词(KeyWords)： 六溴环十二烷;H4细胞;SK-N-AS细胞;神经毒性;脱碘酶

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81773389,81273031);; 大连医科大学教学改革研究项目(DYLX16053)

作者(Author): 郝小吉;尚小娜;于海华;么建萍;韩璐;刘晓晖;李亚晨;邵静;

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DOI:

参考文献(References)：

• [1] Law R J,Covaci A,Harrad S,et al. Levels and trends of PBDEs and HBCDs in the global environment:Status at the end of 2012[J]. Environment International,2014,65:147-158
• [2] Covaci A,Gerecke A C,Law R J,et al. Hexabromocyclododecanes(HBCDs)in the environment and humans:A review[J]. Environmental Science&Technology,2006,40(12):3679-3688
• [3] Aylward L L,Hays S M. Biomonitoring-based risk assessment for hexabromocyclododecane(HBCD)[J].The International Journal of Hygiene and Environmental Health,2011,214:179-187
• [4] Cantón R F,Peijnenburg A A,Hoogenboom R L,et al.Subacute effects of hexabromocyclododecane(HBCD)on hepatic gene expression profiles in rats[J]. Toxicology and Applied Pharmacology,2008,231:267-272
• [5] van der Ven L T,Verhoef A,van de Kuil T,et al. A28-day oral dose toxicity study enhanced to detect endocrine effects of hexabromocyclododecane in Wistar rats[J]. Toxicological Sciences,2006,94:281-292
• [6] Kim U J,Oh J E. Tetrabromobisphenol A and hexabromocyclododecane flame retardants in infant-mother paired serum samples,and their relationships with thyroid hormones and environmental factors[J]. Environmental Pollution,2014,184:193-200
• [7] Lilienthal H,van der Ven L T,Piersma A H,et al.Effects of the brominated flame retardant hexabromocyclododecane(HBCD)on dopamine-dependent behavior and brainstem auditory evoked potentials in a one-generation reproduction study in Wistar rats[J]. Toxicology Letters,2009,185:63-72
• [8] Saegusa Y,Fujimoto H,Woo G H,et al. Transient aberration of neuronal development in the hippocampal dentate gyrus after developmental exposure to brominated flame retardants in rats[J]. Archives of Toxicology,2012,86:1431-1442
• [9] Dentice M,Marsili A,Zavacki A M,et al. The deiodinases and the control of intracellular thyroid hormone signaling during cellular differentiation[J]. Biochimica et Biophysica Acta,2013,1830(7):3937-3945
• [10] Bocco B M,Werneck-de-Castro J P,Oliveira K C,et al. Type 2 deiodinase disruption in astrocytes results in anxiety-depressive-like behavior in male mice[J]. Endocrinology,2016,157(9):3682-3695
• [11] Hernandez A,Morte B,Belinchón M M,et al. Critical role of types 2 and 3 deiodinases in the negative regulation of gene expression by T3 in the mouse cerebral cortex[J]. Endocrinology,2012,153(6):2919-2928
• [12] Martinez M E,Charalambous M,Saferali A,et al. Genomic imprinting variations in the mouse type 3 deiodinase gene between tissues and brain regions[J]. Molecular Endocrinology,2014,28(11):1875-1886
• [13] Darras V M,Van Herck S L. Iodothyronine deiodinase structure and function:From ascidians to humans[J].Journal of Endocrinology,2012,215(2):189-206
• [14] Galton V A,de Waard E,Parlow A F,et al. Life without the iodothyronine deiodinases[J]. Endocrinology,2014,155(10):4081-4087
• [15] Ortiga-Carvalho T M,Chiamolera M I,Pazos-Moura C C,et al. Hypothalamus-pituitary-thyroid axis[J]. Comprehensive Physiology,2016,6(3):1387-1428
• [16] Scholz S,Mayer I. Molecular biomarkers of endocrine disruption in small model fish[J]. Molecular and Cellular Endocrinology,2008,293:57-70
• [17] 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,2010,97:226-233
• [18]钱波，王健，朱春艳，等．甲状腺激素脱碘酶在孕期BDE209暴露致子代小鼠神经发育毒性中的潜在作用[J]．生态毒理学报，2014,9(3):459-466Qian B,Wang J,Zhu C Y,et al. The role of thyroid hormone deiodinase in developmental neurotoxicity in newborn mice mediated by decabromodiphenyl ether(BDE209)exposure during pregnancy and lactation[J]. Asian Journal of Ecotoxicology,2014,9(3):459-466(in Chinese)
• [19]冀秀玲，刘洋，刘芳，等．六溴环十二烷转甲状腺素蛋白结合活性及其发育期暴露的甲状腺激素干扰效应研究[J]．环境科学，2009,31(9):2191-2195Ji X L,Liu Y,Liu F,et al. Transthyretin-binding activity of hexabromocyclododecanes(HBCDs)and its thyroid hormone disrupting effects after developmental exposure[J]. Environmental Science,2009,31(9):2191-2195(in Chinese)
• [20] Palace V P,Pleskach K,Halldorson T,et al. Biotransformation enzymes and thyroid axis disruption in juvenile rainbow trout(Oncorhynchus mykiss)exposed to hexabromocyclododecane diastereoisomers[J]. Environmental Science&Technology,2008,42(6):1967-1972
• [21]刘园园，冀秀玲，沈群辉，等．六溴环十二烷及其复合污染脑发育期暴露对大鼠甲状腺激素代谢过程的影响[J]．生态环境学报，2012,21(9):1614-1619Liu Y Y,Ji X L,Shen Q H,et al. Disrupting effects of hexabromocyclododecane(HBCD)and its complex compounds on rat thyroid hormone metabolism after developmental exposure[J]. Ecology and Environmental Sciences,2012,21(9):1614-1619(in Chinese)
• [22] García G C,Jeziorski M C,Valverde R C,et al. Effects of iodothyronines on the hepatic outer-ring deiodination pathways in killifish[J]. General and Comparative Endocrinology,2004,135:209-210
• [23] Germer S,Piersma A H,van der Ven L,et al. Subacute effects of the brominated flame retardants hexabromocyclododecane and tetrabromobisphenol-A on hepatic cytochrome P450 levels in rats[J]. Toxicology,2006,218:229-236
• [24] Legler J. New insights into the endocrine disrupting effects of brominated flame retardants[J]. Chemosphere,2008,73:216-222
• [25] Shao J,White C C,Dabrowski M J,et al. The role of mitochondrial and oxidative injury in BDE 47 toxicity to human fetal liver hematopoietic stem cells[J]. Toxicological Science,2008,101(1):81-90
• [26] Shao J,Dabrowski M J,White C C,et al. Flow cytometric analysis of BDE 47 mediated injury to rainbow trout gill epithelial cells[J]. Aquatic Toxicology,2010,97(1):42-50
• [27] Liu X,Wang J,Lu C,et al. The role of lysosomes in BDE 47-mediated activation of mitochondrial apoptotic pathway in Hep G2 cells[J]. Chemosphere,2015,124:10-21
• [28] Franco R,Schoneveld O,Georgakilas A G,et al. Oxidative stress,DNA methylation and carcinogenesis[J].Cancer Letters,2008,266(1):6-11
• [29] Gilbert M E,Lasley S M. Developmental thyroid hormone insufficiency and brain development:A role for brain-derived neurotrophic factor(BDNF)?[J]. Neuroscience,2013,239:253-270
• [30] Wei Z,Liao J,Qi F,et al. Evidence for the contribution of BDNF-TrkB signal strength in neurogenesis:An organotypic study[J]. Neuroscience Letters, 2015,606:48-52
• [31] Pajenda G,Hercher D,Márton G,et al. Spatiotemporally limited BDNF and GDNF overexpression rescues motoneurons destined to die and induces elongative axon growth[J]. Experimental Neurology,2014,261:367-376
• [32] Borghesani P R,Peyrin J M,Klein R,et al. BDNF stimulates migration of cerebellar granule cells[J]. Development,2002,129:1435-1442
• [33] Sui L,Li B M. Effects of perinatal hypothyroidism on regulation of reelin and brain-derived neurotrophic factor gene expression in rat hippocampus:Role of DNA methylation and histone acetylation[J]. Steroids,2010,75(12):988-997
• [34] Ibhazehiebo K,Iwasaki T,Xu M,et al. Brain-derived neurotrophic factor(BDNF)ameliorates the suppression of thyroid hormone-induced granule cell neurite extension by hexabromocyclododecane(HBCD)[J]. Neuroscience Letters,2011,493(1-2):1-7