本文主要研究内容
作者李栋（2019）在《2,4-二氯酚影响性激素水平致斑马鱼雌性化》一文中研究指出：2,4-二氯酚(2,4-dichlorophenol,2,4-DCP)作为一种常见工业原料,在工农业生产过程中被大量排入水环境,已成为最常见的氯酚类污染物之一。研究表明2,4-DCP是一种内分泌干扰物,具有潜在的雌激素效应,能够改变生物体内性激素水平和性激素合成相关基因表达。而对2,4-DCP是否能够影响鱼类的性别分化却少见报道。本研究以斑马鱼(Danio rerio)为实验动物,以不同浓度2,4-DCP对20~40 dpf(性别分化关键时期)斑马鱼进行染毒,通过组织切片、酶联免疫、实时荧光定量和重亚硫酸盐修饰测序法等方法检测各浓度2,4-DCP对雌雄比例、性激素水平、基因表达和DNA甲基化水平等的影响。在此基础上,通过添加芳香化酶抑制剂法倔唑(Fadrozole)来抑制雌二醇(E2)的合成,研究2,4-DCP是否通过影响E2的合成引起斑马鱼的雌性化,初步探讨2,4-DCP改变斑马鱼性比的机制,为氯酚类污染物对鱼类性别分化的影响提供新的资料。通过研究,本论文获得了以下结果:1.不同浓度(80和160μg/L)2,4-DCP暴露和17β-E2(5 ng/L)均可引起雌性斑马鱼的比例明显上升,导致斑马鱼雌性化。2.80μg/L 2,4-DCP和17β-E2均可上调卵黄蛋白原基因(vtg)表达和卵黄蛋白原蛋白(VTG)水平,证实2,4-DCP具有雌激素效应。3.不同浓度2,4-DCP暴露均导致鱼体内11-酮基睾酮(11-KT)水平下调,而80μg/L 2,4-DCP可引起E2水平上调,且2,4-DCP能上调鱼体内的E2/11-KT比值,表明2,4-DCP能够影响斑马鱼雌雄激素的平衡。值得注意的是80μg/L2,4-DCP还可导致雌酮(E1)和雌三醇(E3)水平上调,以及睾酮(T)水平下调,表明E2水平的增加很可能是由于2,4-DCP促进了E2的合成所致。4.与2,4-DCP单独处理相比,法倔唑与各浓度2,4-DCP联合处理均可极显著下调雌性斑马鱼的比例,导致斑马鱼性别比例明显偏向雄性。与2,4-DCP单独处理相比,法倔唑与80μg/L 2,4-DCP联合处理均可明显降低E2的水平。此外,法倔唑与17β-E2联合处理虽也导致斑马鱼雄性化,但雌性比例较法倔唑与各浓度2,4-DCP联合处理组更高。上述结果表明雌激素E2在2,4-DCP引起斑马鱼雌性化的效应中发挥着重要作用。5.2,4-DCP暴露导致斑马鱼的性激素合成相关基因cyp19a1a、cyp19a1b、11β-hsd和17β-hsd的表达模式改变。两个浓度的2,4-DCP均引起cyp19a1a表达上调,提示cyp19a1a可能在2,4-DCP促进E2合成的过程中发挥重要调控作用。另外,160μg/L 2,4-DCP上调11β-hsd和17β-hsd表达,而cyp11b2表达在不同浓度2,4-DCP暴露后均无明显变化,提示还有其他机制参与了2,4-DCP对T和11-KT水平的调控,需要进一步探究。此外,与单独2,4-DCP处理相比,法倔唑与2,4-DCP联合处理下调cyp19a1b表达量,而其它基因表达变化则无明显规律。6.2,4-DCP可引起cyp19a1a启动子区上游61 bp处位点(CpG-61)的去甲基化,并具有一定的浓度依赖效应。另外,CpG-61位点附近存在一个可结合雌激素相关受体(Estrogen-related receptors,ESRR)的顺式作用元件。这表明2,4-DCP可通过引起CpG-61位点的去甲基化上调cyp19a1a表达。而对cyp19a1b而言,各浓度2,4-DCP均未引起启动子区域甲基化水平的明显变化,提示2,4-DCP引起的cyp19a1b表达上调可能与DNA甲基化或去甲基化调控无关。综上所述,2,4-DCP可以通过上调斑马鱼cyp19a1a的表达来促进雌激素的合成,打破了雌雄激素间的平衡,进而导致斑马鱼性别比例明显偏向雌性。其中DNA去甲基化参与2,4-DCP对cyp19a1a的表达调控。而2,4-DCP是否通过影响去甲基转移酶TETs(Ten-eleven translocation proteins)表达引起DNA的去甲基化还需要进一步探究。总之,本研究探讨了2,4-DCP暴露对斑马鱼性别分化的毒性效应及机制,为氯酚类污染物对鱼类性别分化的影响提供了新的证据和资料,有望为人类认识氯酚对水生生物的危害提供理论依据。
Abstract
2,4-er lv fen (2,4-dichlorophenol,2,4-DCP)zuo wei yi chong chang jian gong ye yuan liao ,zai gong nong ye sheng chan guo cheng zhong bei da liang pai ru shui huan jing ,yi cheng wei zui chang jian de lv fen lei wu ran wu zhi yi 。yan jiu biao ming 2,4-DCPshi yi chong nei fen bi gan rao wu ,ju you qian zai de ci ji su xiao ying ,neng gou gai bian sheng wu ti nei xing ji su shui ping he xing ji su ge cheng xiang guan ji yin biao da 。er dui 2,4-DCPshi fou neng gou ying xiang yu lei de xing bie fen hua que shao jian bao dao 。ben yan jiu yi ban ma yu (Danio rerio)wei shi yan dong wu ,yi bu tong nong du 2,4-DCPdui 20~40 dpf(xing bie fen hua guan jian shi ji )ban ma yu jin hang ran du ,tong guo zu zhi qie pian 、mei lian mian yi 、shi shi ying guang ding liang he chong ya liu suan yan xiu shi ce xu fa deng fang fa jian ce ge nong du 2,4-DCPdui ci xiong bi li 、xing ji su shui ping 、ji yin biao da he DNAjia ji hua shui ping deng de ying xiang 。zai ci ji chu shang ,tong guo tian jia fang xiang hua mei yi zhi ji fa jue zuo (Fadrozole)lai yi zhi ci er chun (E2)de ge cheng ,yan jiu 2,4-DCPshi fou tong guo ying xiang E2de ge cheng yin qi ban ma yu de ci xing hua ,chu bu tan tao 2,4-DCPgai bian ban ma yu xing bi de ji zhi ,wei lv fen lei wu ran wu dui yu lei xing bie fen hua de ying xiang di gong xin de zi liao 。tong guo yan jiu ,ben lun wen huo de le yi xia jie guo :1.bu tong nong du (80he 160μg/L)2,4-DCPbao lou he 17β-E2(5 ng/L)jun ke yin qi ci xing ban ma yu de bi li ming xian shang sheng ,dao zhi ban ma yu ci xing hua 。2.80μg/L 2,4-DCPhe 17β-E2jun ke shang diao luan huang dan bai yuan ji yin (vtg)biao da he luan huang dan bai yuan dan bai (VTG)shui ping ,zheng shi 2,4-DCPju you ci ji su xiao ying 。3.bu tong nong du 2,4-DCPbao lou jun dao zhi yu ti nei 11-tong ji gao tong (11-KT)shui ping xia diao ,er 80μg/L 2,4-DCPke yin qi E2shui ping shang diao ,ju 2,4-DCPneng shang diao yu ti nei de E2/11-KTbi zhi ,biao ming 2,4-DCPneng gou ying xiang ban ma yu ci xiong ji su de ping heng 。zhi de zhu yi de shi 80μg/L2,4-DCPhai ke dao zhi ci tong (E1)he ci san chun (E3)shui ping shang diao ,yi ji gao tong (T)shui ping xia diao ,biao ming E2shui ping de zeng jia hen ke neng shi you yu 2,4-DCPcu jin le E2de ge cheng suo zhi 。4.yu 2,4-DCPchan du chu li xiang bi ,fa jue zuo yu ge nong du 2,4-DCPlian ge chu li jun ke ji xian zhe xia diao ci xing ban ma yu de bi li ,dao zhi ban ma yu xing bie bi li ming xian pian xiang xiong xing 。yu 2,4-DCPchan du chu li xiang bi ,fa jue zuo yu 80μg/L 2,4-DCPlian ge chu li jun ke ming xian jiang di E2de shui ping 。ci wai ,fa jue zuo yu 17β-E2lian ge chu li sui ye dao zhi ban ma yu xiong xing hua ,dan ci xing bi li jiao fa jue zuo yu ge nong du 2,4-DCPlian ge chu li zu geng gao 。shang shu jie guo biao ming ci ji su E2zai 2,4-DCPyin qi ban ma yu ci xing hua de xiao ying zhong fa hui zhao chong yao zuo yong 。5.2,4-DCPbao lou dao zhi ban ma yu de xing ji su ge cheng xiang guan ji yin cyp19a1a、cyp19a1b、11β-hsdhe 17β-hsdde biao da mo shi gai bian 。liang ge nong du de 2,4-DCPjun yin qi cyp19a1abiao da shang diao ,di shi cyp19a1ake neng zai 2,4-DCPcu jin E2ge cheng de guo cheng zhong fa hui chong yao diao kong zuo yong 。ling wai ,160μg/L 2,4-DCPshang diao 11β-hsdhe 17β-hsdbiao da ,er cyp11b2biao da zai bu tong nong du 2,4-DCPbao lou hou jun mo ming xian bian hua ,di shi hai you ji ta ji zhi can yu le 2,4-DCPdui The 11-KTshui ping de diao kong ,xu yao jin yi bu tan jiu 。ci wai ,yu chan du 2,4-DCPchu li xiang bi ,fa jue zuo yu 2,4-DCPlian ge chu li xia diao cyp19a1bbiao da liang ,er ji ta ji yin biao da bian hua ze mo ming xian gui lv 。6.2,4-DCPke yin qi cyp19a1aqi dong zi ou shang you 61 bpchu wei dian (CpG-61)de qu jia ji hua ,bing ju you yi ding de nong du yi lai xiao ying 。ling wai ,CpG-61wei dian fu jin cun zai yi ge ke jie ge ci ji su xiang guan shou ti (Estrogen-related receptors,ESRR)de shun shi zuo yong yuan jian 。zhe biao ming 2,4-DCPke tong guo yin qi CpG-61wei dian de qu jia ji hua shang diao cyp19a1abiao da 。er dui cyp19a1ber yan ,ge nong du 2,4-DCPjun wei yin qi qi dong zi ou yu jia ji hua shui ping de ming xian bian hua ,di shi 2,4-DCPyin qi de cyp19a1bbiao da shang diao ke neng yu DNAjia ji hua huo qu jia ji hua diao kong mo guan 。zeng shang suo shu ,2,4-DCPke yi tong guo shang diao ban ma yu cyp19a1ade biao da lai cu jin ci ji su de ge cheng ,da po le ci xiong ji su jian de ping heng ,jin er dao zhi ban ma yu xing bie bi li ming xian pian xiang ci xing 。ji zhong DNAqu jia ji hua can yu 2,4-DCPdui cyp19a1ade biao da diao kong 。er 2,4-DCPshi fou tong guo ying xiang qu jia ji zhuai yi mei TETs(Ten-eleven translocation proteins)biao da yin qi DNAde qu jia ji hua hai xu yao jin yi bu tan jiu 。zong zhi ,ben yan jiu tan tao le 2,4-DCPbao lou dui ban ma yu xing bie fen hua de du xing xiao ying ji ji zhi ,wei lv fen lei wu ran wu dui yu lei xing bie fen hua de ying xiang di gong le xin de zheng ju he zi liao ,you wang wei ren lei ren shi lv fen dui shui sheng sheng wu de wei hai di gong li lun yi ju 。
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论文作者分别是来自兰州大学的李栋，发表于刊物兰州大学2019-07-29论文，是一篇关于斑马鱼论文,性别分化论文,性激素论文,性激素合成相关基因论文,去甲基化论文,兰州大学2019-07-29论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自兰州大学2019-07-29论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：斑马鱼论文; 性别分化论文; 性激素论文; 性激素合成相关基因论文; 去甲基化论文; 兰州大学2019-07-29论文;