臺灣博碩士論文加值系統

背景：鄰苯二甲酸酯類(Phthalates, PAEs)中的鄰苯二甲酸二(2-乙基己基)酯(Di(2-ethylhexyl)phthalate, DEHP)為使用量最大的塑化劑，同時也是一種內分泌干擾物，在懷孕期間暴露DEHP會對雄性子代造成生殖危害，且具有超代影響。

目的：探討雌性大鼠於妊娠期間暴露低劑量DEHP，是否對雄性子代造成精子毒性、睪丸毒性與精子DNA甲基化之超代效應。

方法：Sprague-Dawley雌性大鼠(F0)於妊娠期暴露低劑量DEHP 20及200 µg/kg/day，產下雄性子代(F1)，雄性第一子代(F1)於第70天和未暴露之母鼠交配產下第二子代(F2)；雄性F2再與未暴露之母鼠產下第三子代(F3)。只有F0以餵管餵食暴露DEHP，其餘子代均不直接暴露。雄性子代F1、F2、F3測量肛門口至尿道口距離(Anogential Distances, AGD)、肛門與生殖器指數(Anogenital index, AGI)、睪丸和精子功能(包括體重和生殖器官重量)，並對F1、F2和F3的生殖害評估了精子數量、精子活動力、精子形態、線粒體膜電位(Mitochondrial membrane potential, MMP)、精子反應性氧化物質(Reactive Oxygen Species, ROS)生成、DNA片段指數(DNA fragment index, DFI)、睪丸細胞含量分析和血清睪固酮濃度及甲基化轉移酶Dnmt1、Dnmt3a及Dnmt3b。此外，F3進行全基因體亞硫酸鹽定序分析精子DNA甲基化變化，探討低劑量DEHP暴露對雄性大鼠精子功能之表觀基因學超代影響。
 
結果：懷孕期間暴露低劑量DEHP，在F1子代發現20 µg/kg暴露組在生長發育中開眼天數顯著小於控制組，AGD、AGI及精子DFI顯著大於控制組；200 µg/kg暴露組在長牙天數、雄性及雌性仔鼠離乳前體重與雄性仔鼠離乳後體重顯著小於控制組；AGI、精子MMP及精子DFI顯著大於控制組。在F2子代發現20 µg/kg暴露組在精子型態異常中彎尾比例顯著大於控制組；200 µg/kg暴露組精子DFI顯著大於控制組。在F3子代20 µg/kg暴露組精子ROS生成及精子DFI顯著大於控制組，甲基化轉移酶Dnmt1濃度顯著低於控制組，並發現24個高甲基化基因及11個低甲基化基因；200 µg/kg暴露組開眼天數、精子ROS生成及精子DFI顯著大於控制組，並發現27個高甲基化基因及18個低甲基化基因。

結論：懷孕期間暴露低劑量DEHP，可能對其雄性子代大鼠造成生殖相關毒性影響，並經由表觀遺傳學途徑影響第三子代(F3)DNA甲基化改變，造成精子功能之超代效應。

Background：Di(2-ethylhexyl) phthalate (DEHP) is widely used as a plasticizer in the manufacture of plastics. In utero exposure to DEHP may affect male reproductive system and have been shown to have transgenerational effects.

Objectives：The present study was to investigate the low-dose DEHP maternal exposure on reproductive functions and epigenetic transgenerational effects in rat male offspring.

Methods：Pregnant SD rats (F0) were treated through gavage on GD 0 to birth with a vehicle control (corn oil), 20 µg /kg/day DEHP, or 200 µg/kg/day DEHP. Only the F0 generation gestating female was exposed directly. Male rats born to rat from the F1 were labeled the F2. Male rats born to rat from the F2 were labeled the F3. The offspring body weight, anogenital distance (AGD), anogenital index (AGI), tissues weights, sperm count, motility, morphology sperm reactive oxygen species (ROS), mitochondrial membrane potential (MMP), DNA fragmentation index (DFI), DNA content analysis of testis cells, serum testosterone and DNA methyltransferases (Dnmts) were measured for all generations. Whole-genome bisulphite sequencing (WGBS) was also examined to analyze sperm DNA methylation status in the F3 generation.

Results：DEHP exposure at 20 µg/kg affected developmental, AGD, AGI and mean DFI in the F1; sperm morphology in the F2; and ROS, mean DFI, and Dnmt1 in the F3. DEHP exposure at 200 µg/kg affected developmental, body wight, AGI, MMP and mean DFI in the F1; mean DFI in the F2; and ROS and mean DFI in F3. Compared with the control group, 24 and 27 differentially hypermethylated genes were identified in the groups administered F3 20 µg/kg and 200 µg/kg DEHP, respectively. Moreover, 11 and 18 differentially hypomethylated genes were observed in the groups administered 20 µg/kg and 200 µg/kg DEHP.

Conclusion：Pregnant rats exposed to low-dose DEHP might induce reproductive toxicity male offsprings and the epigenetic transgenerational effects in F3.

壹、緒論
1.1 前言
1.2 研究目的

貳、文獻回顧
2.1 環境荷爾蒙
2.2 塑化劑
2.3 鄰苯二甲酸二(2-乙基己基)酯(DEHP)
2.3.1 DEHP之介紹
2.3.2 DEHP之毒物動力學
2.3.3 DEHP之生殖危害
2.4 生殖系統介紹
2.4.1 雄性生殖機能
2.4.2 精子的生成及結構
2.4.3 精子細胞週期及睪丸 DNA 含量
2.5 粒線體功能介紹
2.6 反應性氧化物質
2.6.1 反應性氧化物質介紹
2.6.2 反應性氧化物質與精子功能影響
2.7 超代效應
2.7.1 母系
2.7.2 父系
2.8 表觀遺傳學
2.8.1 DNA甲基化
2.8.2 DNA甲基化轉移酶
2.9 DNA 定序
2.9.1 次世代定序(NGS)
2.9.2 全基因定序(WGS)
2.9.3 全基因體亞硫酸氫鹽定序(WGBS)
2.10 流式細胞儀
2.10.1 基本原理
2.10.2 分析與應用

參、材料與方法
3.1 研究設計
3.2 實驗動物來源
3.3 雄性生殖毒理評估
3.3.1 藥品與試劑
3.3.2 儀器與設備
3.3.3 懸浮液配製
3.3.4 精子數量
3.3.5 精子活動力
3.3.6 精子型態評估
3.3.7 精子染色質結構分析
3.3.8 精子粒線體膜電位分析
3.3.9 精子細胞內超氧陰離子含量分析
3.3.10 精子細胞內過氧化氫含量分析
3.3.11 睪丸細胞DNA含量分析
3.3.12 睪丸組織病理學分析
3.3.13 血清睪固酮
3.3.14 血清尿素氮、肌酐酸及尿酸
3.4 DNA 甲基化分析
3.4.1 精子樣本收集
3.4.2 全基因體亞硫酸鹽定序技術(WGBS)
3.5 DNA 甲基化轉移酶
3.6 統計分析

肆、結果
4.1 雌鼠生育之影響
4.1.1 妊娠期雌鼠體重變化
4.1.2 生長發育指標
4.1.3 離乳前體重觀察
4.1.4 離乳後雄鼠體重觀察
4.1.5 肛門口至尿道口距離(AGD)觀察
4.2 雄性生殖功能影響評估
4.2.1 體重與臟器組織重量分析
4.2.2 精子數量及活動力分析
4.2.3 精子粒線體功能分析
4.2.4 精子細胞反應性氧化物分析
4.2.5 精子染色質DNA結構分析
4.2.6 血清睪固酮
4.2.7 睪丸細胞DNA含量分析
4.2.8 精子型態
4.2.9 睪丸組織病理學
4.2.10 血清尿素氮、肌酐酸及尿酸分析
4.3 第三子代(F3)DNA甲基化
4.3.1 甲基化基因與功能分群
4.3.2 甲基化與劑量效應
4.3.3 甲基化與基因表達
4.4 DNA 甲基化轉移酶
4.5 綜合結果

伍、討論
5.1 生長發育功能影響評估
5.1.1 雌鼠生育影響
5.1.2 各項生長發育指標與雄性子代大鼠體重
5.1.3 尿道口至肛門口距離
5.1.4 雄性子代臟器組織重量
5.2 生殖功能影響評估
5.2.1 雄性子代精子功能之影響評估
5.2.2 DNA 甲基化
5.2.3 DNA甲基化轉移酶
5.3 劑量效應與生殖功能影響評估

陸、結論

柒、參考文獻

捌、研討會發表

玖、附錄

Abdel-Maksoud, F. M., Leasor, K. R., Butzen, K., Braden, T. D. & Akingbemi, B. T. (2015) Prenatal exposures of male rats to the environmental chemicals bisphenol A and di (2-ethylhexyl) phthalate impact the sexual differentiation process. Endocrinology, 156(12), 4672-4683.
Andrade, A. J., Grande, S. W., Talsness, C. E., Grote, K., Golombiewski, A., Sterner-Kock, A. & Chahoud, I. (2006) A dose–response study following in utero and lactational exposure to di-(2-ethylhexyl) phthalate (DEHP): effects on androgenic status, developmental landmarks and testicular histology in male offspring rats. Toxicology, 225(1), 64-74.
Anway, M. D. & Skinner, M. K. (2006) Epigenetic Transgenerational Actions of Endocrine Disruptors. Endocrinology, 147(6), s43-s49. 10.1210/en.2005-1058.
Araki, A., Mitsui, T., Goudarzi, H., Nakajima, T., Miyashita, C., Itoh, S., Sasaki, S., Cho, K., Moriya, K., Shinohara, N., Nonomura, K. & Kishi, R. (2017) Prenatal di(2-ethylhexyl) phthalate exposure and disruption of adrenal androgens and glucocorticoids levels in cord blood: The Hokkaido Study. Science of The Total Environment, 581-582, 297-304.
Arends, R. M., Pasman, J. A., Verweij, K. J., Derks, E. M., Gordon, S. D., Hickie, I., Thomas, N. S., Aliev, F., Zietsch, B. P. & van der Zee, M. D. (2021) Associations between the CADM2 gene, substance use, risky sexual behavior, and self‐control: A phenome‐wide association study. Addiction biology, 26(6), e13015.
Axelsson, J., Rylander, L., Rignell-Hydbom, A., Jönsson, B. A. G., Lindh, C. H. & Giwercman, A. (2015a) Phthalate exposure and reproductive parameters in young men from the general Swedish population. Environment International, 85, 54-60.
Axelsson, J., Rylander, L., Rignell-Hydbom, A., Lindh, C. H., Jönsson, B. A. G. & Giwercman, A. (2015b) Prenatal phthalate exposure and reproductive function in young men. Environmental Research, 138, 264-270.
Banerjee, P. & Bhattacharya, J. (2019) Impact of oxidative stress on infertility, with emphasis on infertility management strategies. Global Journal of Fertility and Research, 4(1), 010-018.
Barakat, R., Lin, P.-C. P., Rattan, S., Brehm, E., Canisso, I. F., Abosalum, M. E., Flaws, J. A., Hess, R. & Ko, C. (2017) Prenatal exposure to DEHP induces premature reproductive senescence in male mice. Toxicological Sciences, 156(1), 96-108.
Barakat, R., Seymore, T., Lin, P.-C. P., Park, C. J. & Ko, C. J. (2019a) Prenatal exposure to an environmentally relevant phthalate mixture disrupts testicular steroidogenesis in adult male mice. Environmental Research, 172, 194-201.
Barakat, R., Seymore, T., Lin, P. P., Park, C. J. & Ko, C. J. (2019b) Prenatal exposure to an environmentally relevant phthalate mixture disrupts testicular steroidogenesis in adult male mice. Environ Res, 172, 194-201. 10.1016/j.envres.2019.02.017.
Bergman, Å., Heindel, J. J., Jobling, S., Kidd, K., Zoeller, T. R. & Organization, W. H. (2013) State of the science of endocrine disrupting chemicals 2012. World Health Organization.
Bloom, M. S., Whitcomb, B. W., Chen, Z., Ye, A., Kannan, K. & Buck Louis, G. M. (2015) Associations between urinary phthalate concentrations and semen quality parameters in a general population. Human Reproduction, 30(11), 2645-2657. 10.1093/humrep/dev219.
Brehm, E. & Flaws, J. A. (2019) Transgenerational Effects of Endocrine-Disrupting Chemicals on Male and Female Reproduction. Endocrinology, 160(6), 1421-1435. 10.1210/en.2019-00034.
Brieger, K., Schiavone, S., Miller, F. J. & Krause, K.-H. (2012) Reactive oxygen species: from health to disease. Swiss medical weekly, 142, w13659.
Bungum, M., Bungum, L. & Giwercman, A. (2011) Sperm chromatin structure assay (SCSA): a tool in diagnosis and treatment of infertility. Asian journal of andrology, 13(1), 69-75. 10.1038/aja.2010.73.
Chédin, F. (2011) Chapter 7 - The DNMT3 Family of Mammalian De Novo DNA Methyltransferases. In: Cheng, X. & Blumenthal, R. M. (eds.) Progress in Molecular Biology and Translational Science. Academic Press.
Chaffey, N. (2003) Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K. and Walter, P. Molecular biology of the cell. 4th edn. Annals of Botany, 91(3), 401-401. 10.1093/aob/mcg023.
Chanjuan Hao, X. C., Jian Guo & , H. X., Xu Ma (2013) Perinatal exposure to diethyl-hexyl-phthalate induces obesity in mice
Chen, J., Wu, S., Wen, S., Shen, L., Peng, J., Yan, C., Cao, X., Zhou, Y., Long, C. & Lin, T. (2015) The mechanism of environmental endocrine disruptors (DEHP) induces epigenetic transgenerational inheritance of cryptorchidism. PloS one, 10(6), e0126403.
Chen, T., Yang, W., Li, Y., Chen, X. & Xu, S. (2011) Mono-(2-ethylhexyl) phthalate impairs neurodevelopment: Inhibition of proliferation and promotion of differentiation in PC12 cells. Toxicology Letters, 201(1), 34-41.
Chung, C.-Y., Sun, Z., Mullokandov, G., Bosch, A., Qadeer, Z. A., Cihan, E., Rapp, Z., Parsons, R., Aguirre-Ghiso, J. A., Farias, E. F., Brown, B. D., Gaspar-Maia, A. & Bernstein, E. (2016) Cbx8 Acts Non-canonically with Wdr5 to Promote Mammary Tumorigenesis. Cell Reports, 16(2), 472-486.
Dagher, J. B., Hahn-Townsend, C. K., Kaimal, A., Mansi, M. A., Henriquez, J. E., Tran, D. G., Laurent, C. R., Bacak, C. J., Buechter, H. E., Cambric, C., Spivey, J., Chuang, Y.-J., Campbell, E. J., Mandal, A., Mohankumar, P. S. & MohanKumar, S. M. J. (2021a) Independent and combined effects of Bisphenol A and Diethylhexyl Phthalate on gestational outcomes and offspring development in Sprague-Dawley rats. Chemosphere, 263, 128307.
Dagher, J. B., Hahn-Townsend, C. K., Kaimal, A., Mansi, M. A., Henriquez, J. E., Tran, D. G., Laurent, C. R., Bacak, C. J., Buechter, H. E., Cambric, C., Spivey, J., Chuang, Y. J., Campbell, E. J., Mandal, A., Mohankumar, P. S. & MohanKumar, S. M. J. (2021b) Independent and combined effects of Bisphenol A and Diethylhexyl Phthalate on gestational outcomes and offspring development in Sprague-Dawley rats. Chemosphere, 263, 128307. 10.1016/j.chemosphere.2020.128307.
Desdoits-Lethimonier, C., Albert, O., Le Bizec, B., Perdu, E., Zalko, D., Courant, F., Lesne, L., Guille, F., Dejucq-Rainsford, N. & Jegou, B. (2012) Human testis steroidogenesis is inhibited by phthalates. Hum Reprod, 27(5), 1451-9. 10.1093/humrep/des069.
Diamanti-Kandarakis, E., Bourguignon, J.-P., Giudice, L. C., Hauser, R., Prins, G. S., Soto, A. M., Zoeller, R. T. & Gore, A. C. (2009) Endocrine-Disrupting Chemicals: An Endocrine Society Scientific Statement. Endocrine Reviews, 30(4), 293-342. 10.1210/er.2009-0002.
Do, R. P., Stahlhut, R. W., Ponzi, D., vom Saal, F. S. & Taylor, J. A. (2012) Non-monotonic dose effects of in utero exposure to di(2-ethylhexyl) phthalate (DEHP) on testicular and serum testosterone and anogenital distance in male mouse fetuses. Reproductive Toxicology, 34(4), 614-621.
Dodé, C. & Hardelin, J.-P. Kallmann syndrome: mutations in the genes encoding prokineticin-2 (PROK2) and prokineticin receptor-2 (PROKR2). Endocrine Abstracts. Bioscientifica.
Dodé, C., Teixeira, L., Levilliers, J., Fouveaut, C., Bouchard, P., Kottler, M.-L., Lespinasse, J., Lienhardt-Roussie, A., Mathieu, M. & Moerman, A. (2006) Kallmann syndrome: mutations in the genes encoding prokineticin-2 and prokineticin receptor-2. PLoS genetics, 2(10), e175.
Doyle, T. J., Bowman, J. L., Windell, V. L., McLean, D. J. & Kim, K. H. (2013) Transgenerational effects of di-(2-ethylhexyl) phthalate on testicular germ cell associations and spermatogonial stem cells in mice. Biol Reprod, 88(5), 112. 10.1095/biolreprod.112.106104.
Ejaredar, M., Nyanza, E. C., Ten Eycke, K. & Dewey, D. (2015) Phthalate exposure and childrens neurodevelopment: A systematic review. Environmental Research, 142, 51-60.
Erenpreiss, J., Spano, M., Erenpreisa, J., Bungum, M. & Giwercman, A. (2006) Sperm chromatin structure and male fertility: biological and clinical aspects. Asian journal of andrology, 8(1), 11-29.
Erkekoglu, P., Rachidi, W., Yuzugullu, O. G., Giray, B., Favier, A., Ozturk, M. & Hincal, F. (2010) Evaluation of cytotoxicity and oxidative DNA damaging effects of di(2-ethylhexyl)-phthalate (DEHP) and mono(2-ethylhexyl)-phthalate (MEHP) on MA-10 Leydig cells and protection by selenium. Toxicology and Applied Pharmacology, 248(1), 52-62.
Fadayomi, I., Sari, S., Kitchen, M., Reynisson, J., Forsyth, N. & Li, W.-W. (2022) Sesquiterpene Lactones Modulated DNA Methylation through Inhibition of DNMTs in Ovarian Cancer Cells. Pharmacological Research - Modern Chinese Medicine, 3, 100074.
Fong, J. P., Lee, F. J., Lu, I. S., Uang, S. N. & Lee, C. C. (2015) Relationship between urinary concentrations of di(2-ethylhexyl) phthalate (DEHP) metabolites and reproductive hormones in polyvinyl chloride production workers. Occup Environ Med, 72(5), 346-53. 10.1136/oemed-2014-102532.
Forzati, F., Federico, A., Pallante, P., Abbate, A., Esposito, F., Malapelle, U., Sepe, R., Palma, G., Troncone, G. & Scarfò, M. (2012) CBX7 is a tumor suppressor in mice and humans. The Journal of clinical investigation, 122(2).
Gadadhar, S., Hirschmugl, T. & Janke, C. (2022) The tubulin code in mammalian sperm development and function. Seminars in Cell & Developmental Biology.
Gao, H.-T., Xu, R., Cao, W.-X., Qian, L.-L., Wang, M., Lu, L., Xu, Q. & Yu, S.-Q. (2017) Effects of six priority controlled phthalate esters with long-term low-dose integrated exposure on male reproductive toxicity in rats. Food and Chemical Toxicology, 101, 94-104.
Garrido-Cardenas, J. A., Garcia-Maroto, F., Alvarez-Bermejo, J. A. & Manzano-Agugliaro, F. (2017) DNA sequencing sensors: an overview. Sensors, 17(3), 588.
Gillooly, J. F., Hein, A. & Damiani, R. (2015) Nuclear DNA content varies with cell size across human cell types. Cold Spring Harbor Perspectives in Biology, 7(7), a019091.
Gore, A. C., Chappell, V. A., Fenton, S. E., Flaws, J. A., Nadal, A., Prins, G. S., Toppari, J. & Zoeller, R. T. (2015) Executive Summary to EDC-2: The Endocrine Society's Second Scientific Statement on Endocrine-Disrupting Chemicals. Endocr Rev, 36(6), 593-602. 10.1210/er.2015-1093.
Han, T. S. & Bouloux, P. M. G. (2012) Chapter 27 - Kallmann Syndrome and Other Causes of Hypothalamic Hypogonadism and Related Development Disorders. In: Fink, G., Pfaff, D. W. & Levine, J. E. (eds.) Handbook of Neuroendocrinology. San Diego: Academic Press.
Hsu, P.-C., Jhong, J.-Y., Huang, L.-P., Lee, K.-H., Chen, H.-P. & Guo, Y.-L. (2021a) Transgenerational effects of Di (2-Ethylhexyl) phthalate on anogenital distance, sperm functions and DNA methylation in rat offspring. International Journal of Molecular Sciences, 22(8), 4131.
Hsu, P.-C., Li, Z.-K., Lai, C.-S., Tseng, L.-H., Lee, C.-W., Cheng, F.-J., Chang, C.-Y. & Chen, J.-R. (2021b) Transgenerational effects of BDE-209 on male reproduction in F3 offspring rats. Chemosphere, 272, 129829.
Hsu, P. C., Jhong, J. Y., Huang, L. P., Lee, K. H., Chen, H. P. & Guo, Y. L. (2021c) Transgenerational Effects of Di(2-Ethylhexyl) Phthalate on Anogenital Distance, Sperm Functions and DNA Methylation in Rat Offspring. Int J Mol Sci, 22(8). 10.3390/ijms22084131.
Huang da, W., Sherman, B. T. & Lempicki, R. A. (2009a) Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res, 37(1), 1-13. 10.1093/nar/gkn923.
Huang da, W., Sherman, B. T. & Lempicki, R. A. (2009b) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc, 4(1), 44-57. 10.1038/nprot.2008.211.
Huang, L.-L., Zhou, B., Ai, S.-H., Yang, P., Chen, Y.-J., Liu, C., Deng, Y.-L., Lu, Q., Miao, X.-P. & Lu, W.-Q. (2018) Prenatal phthalate exposure, birth outcomes and DNA methylation of Alu and LINE-1 repetitive elements: a pilot study in China. Chemosphere, 206, 759-765.
Huang, L.-P., Lee, C.-C., Fan, J.-P., Kuo, P.-H., Shih, T.-S. & Hsu, P.-C. (2014) Urinary metabolites of di (2-ethylhexyl) phthalate relation to sperm motility, reactive oxygen species generation, and apoptosis in polyvinyl chloride workers. International archives of occupational and environmental health, 87(6), 635-646.
Huang, L. P., Lee, C. C., Hsu, P. C. & Shih, T. S. (2011) The association between semen quality in workers and the concentration of di(2-ethylhexyl) phthalate in polyvinyl chloride pellet plant air. Fertil Steril, 96(1), 90-4. 10.1016/j.fertnstert.2011.04.093.
Huen, K., Calafat, A. M., Bradman, A., Yousefi, P., Eskenazi, B. & Holland, N. (2016) Maternal phthalate exposure during pregnancy is associated with DNA methylation of LINE-1 and Alu repetitive elements in Mexican-American children. Environmental research, 148, 55-62.
Jackson, M. J. & McArdle, A. (2011) Age‐related changes in skeletal muscle reactive oxygen species generation and adaptive responses to reactive oxygen species. The Journal of physiology, 589(9), 2139-2145.
Jedynak, P., Tost, J., Calafat, A. M., Bourova-Flin, E., Broséus, L., Busato, F., Forhan, A., Heude, B., Jakobi, M., Schwartz, J., Slama, R., Vaiman, D., Lepeule, J. & Philippat, C. (2022) Pregnancy exposure to phthalates and DNA methylation in male placenta — An epigenome-wide association study. Environment International, 160, 107054.
Joensen, U. N., Frederiksen, H., Jensen, M. B., Lauritsen, M. P., Olesen, I. A., Lassen, T. H., Andersson, A.-M. & Jørgensen, N. (2012) Phthalate excretion pattern and testicular function: a study of 881 healthy Danish men. Environmental health perspectives, 120(10), 1397-1403.
Jurewicz, J., Radwan, M., Sobala, W., Ligocka, D., Radwan, P., Bochenek, M., Hawuła, W., Jakubowski, L. & Hanke, W. (2013) Human urinary phthalate metabolites level and main semen parameters, sperm chromatin structure, sperm aneuploidy and reproductive hormones. Reproductive Toxicology, 42, 232-241.
Kang, L., Chen, J., Wang, J., Zhao, T., Wei, Y., Wu, Y., Han, L., Zheng, X., Shen, L. & Long, C. (2022) Multiple transcriptomic profiling: potential novel metabolism-related genes predict prepubertal testis damage caused by DEHP exposure. Environmental Science and Pollution Research, 29(9), 13478-13490.
Kim, S., Kang, S., Lee, G., Lee, S., Jo, A., Kwak, K., Kim, D., Koh, D., Kho, Y. L. & Kim, S. (2014) Urinary phthalate metabolites among elementary school children of Korea: sources, risks, and their association with oxidative stress marker. Science of the Total Environment, 472, 49-55.
Knoblaugh, S. E., True, L., Tretiakova, M. & Hukkanen, R. R. (2018) 18 - Male Reproductive System. In: Treuting, P. M., Dintzis, S. M. & Montine, K. S. (eds.) Comparative Anatomy and Histology (Second Edition). San Diego: Academic Press.
Koch, H. M., Bolt, H. M., Preuss, R. & Angerer, J. (2005) New metabolites of di (2-ethylhexyl) phthalate (DEHP) in human urine and serum after single oral doses of deuterium-labelled DEHP. Archives of toxicology, 79(7), 367-376.
Koch, H. M., Rossbach, B., Drexler, H. & Angerer, J. (2003) Internal exposure of the general population to DEHP and other phthalates—determination of secondary and primary phthalate monoester metabolites in urine. Environmental Research, 93(2), 177-185.
Lee, D. H. & Jacobs, D. R., Jr. (2015) Methodological issues in human studies of endocrine disrupting chemicals. Rev Endocr Metab Disord, 16(4), 289-97. 10.1007/s11154-016-9340-9.
Lee, I., Alakeel, R., Kim, S., Al-Sheikh, Y. A., Al-Mandeel, H., Alyousef, A. A., Kho, Y. & Choi, K. (2019) Urinary phthalate metabolites among children in Saudi Arabia: Occurrences, risks, and their association with oxidative stress markers. Science of the Total Environment, 654, 1350-1357.
Lin, P.-H., Jian, C.-Y., Chou, J.-C., Chen, C.-W., Chen, C.-C., Soong, C., Hu, S., Lieu, F.-K., Wang, P. S. & Wang, S.-W. (2016) Induction of renal senescence marker protein-30 (SMP30) expression by testosterone and its contribution to urinary calcium absorption in male rats. Scientific reports, 6(1), 1-12.
Liou, Y. J., Chen, M. L., Wang, Y. C., Chen, J. Y., Liao, D. L., Bai, Y. M., Lin, C. C., Chen, T. T., Mo, G. H. & Lai, I. C. (2009) Analysis of genetic variations in the RGS9 gene and antipsychotic‐induced tardive dyskinesia in schizophrenia. American Journal of Medical Genetics Part B: Neuropsychiatric Genetics, 150(2), 239-242.
Lite, C., Raja, G. L., Juliet, M., Sridhar, V. V., Subhashree, K. D., Kumar, P., Chakraborty, P. & Arockiaraj, J. (2022) In utero exposure to endocrine-disrupting chemicals, maternal factors and alterations in the epigenetic landscape underlying later-life health effects. Environmental Toxicology and Pharmacology, 89, 103779.
Lopez-Rodriguez, D., Franssen, D., Heger, S. & Parent, A.-S. (2021) Endocrine-disrupting chemicals and their effects on puberty. Best Practice & Research Clinical Endocrinology & Metabolism, 35(5), 101579.
Lowe, J. S. & Anderson, P. G. (2014) Stevens & Lowe's Human Histology E-Book: With STUDENT CONSULT Online Access. Elsevier Health Sciences.
Lymperi, S. & Giwercman, A. (2018) Endocrine disruptors and testicular function. Metabolism, 86, 79-90.
Massart, A., Lissens, W., Tournaye, H. & Stouffs, K. (2012) Genetic causes of spermatogenic failure. Asian journal of andrology, 14(1), 40.
McBride, H. M., Neuspiel, M. & Wasiak, S. (2006) Mitochondria: More Than Just a Powerhouse. Current Biology, 16(14), R551-R560.
Meltzer, D., Martinez–Arguelles, D. B., Campioli, E., Lee, S. & Papadopoulos, V. (2015) In utero exposure to the endocrine disruptor di (2-ethylhexyl) phthalate targets ovarian theca cells and steroidogenesis in the adult female rat. Reproductive toxicology, 51, 47-56.
Moraes, C. R. & Meyers, S. (2018) The sperm mitochondrion: Organelle of many functions. Animal Reproduction Science, 194, 71-80.
Morris, J., Bailey, M. E. S., Baldassarre, D., Cullen, B., de Faire, U., Ferguson, A., Gigante, B., Giral, P., Goel, A., Graham, N., Hamsten, A., Humphries, S. E., Johnston, K. J. A., Lyall, D. M., Lyall, L. M., Sennblad, B., Silveira, A., Smit, A. J., Tremoli, E., Veglia, F., Ward, J., Watkins, H., Smith, D. J. & Strawbridge, R. J. (2019) Genetic variation in CADM2 as a link between psychological traits and obesity. Sci Rep, 9(1), 7339. 10.1038/s41598-019-43861-9.
Muerköster, A.-P., Frederiksen, H., Juul, A., Andersson, A.-M., Jensen, R. C., Glintborg, D., Kyhl, H. B., Andersen, M. S., Timmermann, C. A. G. & Jensen, T. K. (2020) Maternal phthalate exposure associated with decreased testosterone/LH ratio in male offspring during mini-puberty. Odense Child Cohort. Environment International, 144, 106025.
Okahisa, Y., Kodama, M., Takaki, M., Inada, T., Uchimura, N., Yamada, M., Iwata, N., Iyo, M., Sora, I. & Ozaki, N. (2011) Association between the regulator of G-protein signaling 9 gene and patients with methamphetamine use disorder and schizophrenia. Current Neuropharmacology, 9(1), 190-194.
Oudir, M., Chader, H., Bouzid, B., Bendisari, K., Latreche, B., Boudalia, S. & Iguer-Ouada, M. (2018) Male rat exposure to low dose of di (2-ethylhexyl) phthalate during pre-pubertal, pubertal and post-pubertal periods: Impact on sperm count, gonad histology and testosterone secretion. Reproductive Toxicology, 75, 33-39.
Pan, Y., Jing, J., Yeung, L. W. Y., Sheng, N., Zhang, H., Yao, B. & Dai, J. (2016) Associations of urinary 5-methyl-2′-deoxycytidine and 5-hydroxymethyl-2′-deoxycytidine with phthalate exposure and semen quality in 562 Chinese adult men. Environment International, 94, 583-590.
Prados, J., Stenz, L., Somm, E., Stouder, C., Dayer, A. & Paoloni-Giacobino, A. (2015) Prenatal exposure to DEHP affects spermatogenesis and sperm DNA methylation in a strain-dependent manner. PloS one, 10(8), e0132136.
Rajesh, P. & Balasubramanian, K. (2015) Gestational exposure to di (2-ethylhexyl) phthalate (DEHP) impairs pancreatic β-cell function in F1 rat offspring. Toxicology Letters, 232(1), 46-57.
Rattan, S., Brehm, E., Gao, L. & Flaws, J. A. (2018a) Di (2-ethylhexyl) phthalate exposure during prenatal development causes adverse transgenerational effects on female fertility in mice. Toxicological Sciences, 163(2), 420-429.
Rattan, S., Brehm, E., Gao, L. & Flaws, J. A. (2018b) Di(2-Ethylhexyl) Phthalate Exposure During Prenatal Development Causes Adverse Transgenerational Effects on Female Fertility in Mice. Toxicological Sciences, 163(2), 420-429. 10.1093/toxsci/kfy042.
Robaire, B., Delbes, G., Head, J. A., Marlatt, V. L., Martyniuk, C. J., Reynaud, S., Trudeau, V. L. & Mennigen, J. A. (2022) A cross-species comparative approach to assessing multi- and transgenerational effects of endocrine disrupting chemicals. Environmental Research, 204, 112063.
Rocha, B. A., Asimakopoulos, A. G., Barbosa, F. & Kannan, K. (2017) Urinary concentrations of 25 phthalate metabolites in Brazilian children and their association with oxidative DNA damage. Science of The Total Environment, 586, 152-162.
Rodprasert, W., Main, K. M., Toppari, J. & Virtanen, H. E. (2019) Associations between male reproductive health and exposure to endocrine-disrupting chemicals. Current Opinion in Endocrine and Metabolic Research, 7, 49-61.
Sanger, F., Air, G. M., Barrell, B. G., Brown, N. L., Coulson, A. R., Fiddes, J. C., Hutchison, C., Slocombe, P. M. & Smith, M. (1977a) Nucleotide sequence of bacteriophage φX174 DNA. nature, 265(5596), 687-695.
Sanger, F., Nicklen, S. & Coulson, A. R. (1977b) DNA sequencing with chain-terminating inhibitors. Proceedings of the national academy of sciences, 74(12), 5463-5467.
Sarfati, J., Dodé, C. & Young, J. (2010) Kallmann syndrome caused by mutations in the PROK2 and PROKR2 genes: pathophysiology and genotype-phenotype correlations. Kallmann syndrome and hypogonadotropic hypogonadism, 39, 121-132.
Schaefer, C. B., Ooi, S. K., Bestor, T. H. & Bourc'his, D. (2007) Epigenetic decisions in mammalian germ cells. science, 316(5823), 398-399.
Sekaran, S. & Jagadeesan, A. (2015) In utero exposure to phthalate downregulates critical genes in Leydig cells of F1 male progeny. Journal of cellular biochemistry, 116(7), 1466-1477.
Shinjo, K., Yamashita, Y., Yamamoto, E., Akatsuka, S., Uno, N., Kamiya, A., Niimi, K., Sakaguchi, Y., Nagasaka, T. & Takahashi, T. (2014) Expression of chromobox homolog 7 (CBX7) is associated with poor prognosis in ovarian clear cell adenocarcinoma via TRAIL‐induced apoptotic pathway regulation. International journal of cancer, 135(2), 308-318.
Skinner, M. K. (2007) Endocrine disruptors and epigenetic transgenerational disease etiology. Pediatric research, 61(7), 48-50.
Sobarzo, C. M., Rosana, N. d. M., Livia, L., Berta, D. & Schteingart, H. F. (2015) Mono-(2-ethylhexyl) phthalate (MEHP) affects intercellular junctions of Sertoli cell: A potential role of oxidative stress. Reproductive Toxicology, 58, 203-212.
Specht, I. O., Toft, G., Hougaard, K. S., Lindh, C. H., Lenters, V., Jönsson, B. A. G., Heederik, D., Giwercman, A. & Bonde, J. P. E. (2014) Associations between serum phthalates and biomarkers of reproductive function in 589 adult men. Environment International, 66, 146-156.
Spencer, H. G. (2016) Epigenetic Inheritance. In: Kliman, R. M. (ed.) Encyclopedia of Evolutionary Biology. Oxford: Academic Press.
Stirzaker, C., Taberlay, P. C., Statham, A. L. & Clark, S. J. (2014) Mining cancer methylomes: prospects and challenges. Trends in Genetics, 30(2), 75-84.
Sun, P.-P., Lai, C.-S., Hung, C.-J., Dhaiveegan, P., Tsai, M.-L., Chiu, C.-L. & Fang, J.-M. (2021) Subchronic oral toxicity evaluation of gold nanoparticles in male and female mice. Heliyon, 7(3), e06577.
Tang, J., Wang, G., Zhang, M., Li, F.-y., Sang, Y., Wang, B., Hu, K., Wu, Y., Luo, R. & Liao, D. (2014) Paradoxical role of CBX8 in proliferation and metastasis of colorectal cancer. Oncotarget, 5(21), 10778.
Tian, M., Liu, L., Zhang, J., Huang, Q. & Shen, H. (2019) Positive association of low-level environmental phthalate exposure with sperm motility was mediated by DNA methylation: A pilot study. Chemosphere, 220, 459-467.
Toshimori, K. & Eddy, E. M. (2015) Chapter 3 - The Spermatozoon. In: Plant, T. M. & Zeleznik, A. J. (eds.) Knobil and Neill's Physiology of Reproduction (Fourth Edition). San Diego: Academic Press.
Tsai, Y.-A., Lin, C.-L., Hou, J.-W., Huang, P.-C., Lee, M.-C., Chen, B.-H., Wu, M.-T., Chen, C.-C., Wang, S.-L., Lee, C.-C., Hsiung, C. A. & Chen, M.-L. (2016) Effects of high di(2-ethylhexyl) phthalate (DEHP) exposure due to tainted food intake on pre-pubertal growth characteristics in a Taiwanese population. Environmental Research, 149, 197-205.
Tsatsakis, A. M., Katsikantami, I., Kalantzi, O.-I., Sevim, Ç., Tsarouhas, K., Sarigiannis, D., Tzatzarakis, M. N. & Rizos, A. K. (2019) Phthalates: Exposure and Health Effects. In: Nriagu, J. (ed.) Encyclopedia of Environmental Health (Second Edition). Oxford: Elsevier.
Uysal, F., Akkoyunlu, G. & Ozturk, S. (2016) DNA methyltransferases exhibit dynamic expression during spermatogenesis. Reproductive BioMedicine Online, 33(6), 690-702.
Van Cauwenbergh, O., Di Serafino, A., Tytgat, J. & Soubry, A. (2020) Transgenerational epigenetic effects from male exposure to endocrine-disrupting compounds: a systematic review on research in mammals. Clinical epigenetics, 12(1), 1-23.
Vandenberg, L. N., Colborn, T., Hayes, T. B., Heindel, J. J., Jacobs, D. R., Jr., Lee, D.-H., Shioda, T., Soto, A. M., vom Saal, F. S., Welshons, W. V., Zoeller, R. T. & Myers, J. P. (2012a) Hormones and Endocrine-Disrupting Chemicals: Low-Dose Effects and Nonmonotonic Dose Responses. Endocrine Reviews, 33(3), 378-455. 10.1210/er.2011-1050.
Vandenberg, L. N., Colborn, T., Hayes, T. B., Heindel, J. J., Jacobs Jr, D. R., Lee, D.-H., Shioda, T., Soto, A. M., vom Saal, F. S. & Welshons, W. V. (2012b) Hormones and endocrine-disrupting chemicals: low-dose effects and nonmonotonic dose responses. Endocrine reviews, 33(3), 378-455.
Vaz, C. V., Maia, C. J., Marques, R., Gomes, I. M., Correia, S., Alves, M. G., Cavaco, J. E., Oliveira, P. F. & Socorro, S. (2014) Regucalcin is an androgen‐target gene in the rat prostate modulating cell‐cycle and apoptotic pathways. The Prostate, 74(12), 1189-1198.
Vaz, C. V., Marques, R., Cardoso, H. J., Maia, C. J. & Socorro, S. (2016) Suppressed glycolytic metabolism in the prostate of transgenic rats overexpressing calcium-binding protein regucalcin underpins reduced cell proliferation. Transgenic research, 25(2), 139-148.
Wang, Y.-X., You, L., Zeng, Q., Sun, Y., Huang, Y.-H., Wang, C., Wang, P., Cao, W.-C., Yang, P., Li, Y.-F. & Lu, W.-Q. (2015) Phthalate exposure and human semen quality: Results from an infertility clinic in China. Environmental Research, 142, 1-9.
Wright, C., Milne, S. & Leeson, H. (2014) Sperm DNA damage caused by oxidative stress: modifiable clinical, lifestyle and nutritional factors in male infertility. Reproductive BioMedicine Online, 28(6), 684-703.
Wu, S., Zhu, J., Li, Y., Lin, T., Gan, L., Yuan, X., Xiong, J., Liu, X., Xu, M. & Zhao, D. (2010) Dynamic epigenetic changes involved in testicular toxicity induced by di‐2‐(ethylhexyl) phthalate in mice. Basic & clinical pharmacology & toxicology, 106(2), 118-123.
Xin, F., Susiarjo, M. & Bartolomei, M. S. (2015a) Multigenerational and transgenerational effects of endocrine disrupting chemicals: A role for altered epigenetic regulation? Seminars in Cell & Developmental Biology, 43, 66-75.
Xin, F., Susiarjo, M. & Bartolomei, M. S. Multigenerational and transgenerational effects of endocrine disrupting chemicals: A role for altered epigenetic regulation? , Seminars in cell & developmental biology. Elsevier, 66-75.
Yao, P.-L., Lin, Y.-C. & Richburg, J. H. (2010) Mono-(2-Ethylhexyl) Phthalate-Induced Disruption of Junctional Complexes in the Seminiferous Epithelium of the Rodent Testis Is Mediated by MMP21. Biology of Reproduction, 82(3), 516-527. 10.1095/biolreprod.109.080374.
Yin, L.-L., Geng, X.-C. & Zhu, X.-Z. (2011) The involvement of RGS9 in l-3, 4-dihydroxyphenylalanine-induced dyskinesias in unilateral 6-OHDA lesion rat model. Brain research bulletin, 86(5-6), 367-372.
You, H. H. & Song, G. (2021) Review of endocrine disruptors on male and female reproductive systems. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 244, 109002.
Yuan, Y.-Z., Ye, C., Sun, J.-H., Hu, M.-Y., Huo, S.-J., Zhu, Y.-T., Xiang, S.-Y. & Yu, S.-Q. (2022) Toxicokinetics of mono-(2-ethylhexyl) phthalate with low-dose exposure applying fluorescence tracing technique. Toxicology and Applied Pharmacology, 434, 115814.
Zhang, Y., Wang, S., Zhao, T., Yang, L., Guo, S., Shi, Y., Zhang, X., Zhou, L. & Ye, L. (2019) Mono-2-ethylhexyl phthalate (MEHP) promoted lipid accumulation via JAK2/STAT5 and aggravated oxidative stress in BRL-3A cells. Ecotoxicology and Environmental Safety, 184, 109611.
Zhao, Y., Chen, J., Wang, X., Song, Q., Xu, H.-H. & Zhang, Y.-H. (2016) Third trimester phthalate exposure is associated with DNA methylation of growth-related genes in human placenta. Scientific reports, 6(1), 1-8.
Zhou, C., Gao, L. & Flaws, J. A. (2017) Exposure to an environmentally relevant phthalate mixture causes transgenerational effects on female reproduction in mice. Endocrinology, 158(6), 1739-1754.