本文選題：內(nèi)分泌干擾物　切入點：甾體激素受體　出處：《浙江大學》2017年碩士論文

【摘要】：中國的土壤污染呈越來越嚴重的趨勢。我國當前對土壤污染物的甄別和鑒定主要是基于現(xiàn)有的土壤環(huán)境質(zhì)量標準和環(huán)境安全值,但是隨著工農(nóng)業(yè)高速發(fā)展,污染物展現(xiàn)出種類多、來源廣的特點,現(xiàn)有的相關(guān)規(guī)定和標準中關(guān)于污染物的基本毒性數(shù)據(jù)有限甚至缺失、針對污染物的有效且準確的化學檢測方法缺乏,因此已漸漸不能滿足需要,給現(xiàn)代環(huán)境污染的管理和控制帶來了困難。以往對土壤污染的研究聚焦于污染物的時空分布,對復合污染引發(fā)的生態(tài)毒理效應(yīng)的關(guān)注不高。然而環(huán)境中污染物造成的生態(tài)影響和健康風險往往是長期低劑量暴露的結(jié)果,簡單的通過測定污染物濃度來評價其危害和影響是不夠準確的。因此,有必要采用生物學的方法綜合評價土壤污染情況。我們前期的研究表明,中國農(nóng)田土壤中檢出許多典型的環(huán)境內(nèi)分泌干擾物(Endocrine Disrupting Chemicals,EDCs),如有機氯殺蟲劑、多氯聯(lián)苯化合物(PCBs)、多環(huán)芳烴化合物(PAHs)以及鄰苯二甲酸酯類(PAEs)等。這些EDCs可通過模擬內(nèi)源性甾體激素與其受體結(jié)合,產(chǎn)生類激素活性或抗激素活性。甾體激素受體包括雌激素受體(Estrogen Receptor,ER)、雄激素受體(Androgen Receptor,AR)、孕激素受體(Progesterone Receptor,PR)、糖皮質(zhì)激素受體(Glucocorticoid Receptor,GR)和鹽皮質(zhì)激素受體(Mineralocorticoid Receptor,MR)。已有大量研究表明,EDCs可與ER、AR作用,然而關(guān)于EDCs與PR、GR、MR作用的研究相對較少。土壤環(huán)境中還存在很多未知的污染物,生物學的方法評價土壤提取物的內(nèi)分泌干擾效應(yīng),可綜合評價包含未知污染物的總體內(nèi)分泌干擾效應(yīng)情況,因此更利于了解土壤中EDCs的生態(tài)風險。本研究采集了全國31個省或地區(qū)共122個農(nóng)田表層土壤,經(jīng)索氏法提取土壤中有機污染物,利用雙熒光素酶報告基因系統(tǒng)檢測了土壤提取物通過ER、AR、PR、GR以及MR介導的類/抗甾體激素活性,闡明了EDCs在我國農(nóng)田土壤中的分布特征及生態(tài)風險情況。本研究結(jié)果表明:(1)79%土壤提取物樣品具有類雌激素活性,雌二醇當量(EEQ)的范圍為0～7.34 ng EEQ kg~(-1) soil(d.w.),平均值為1.94 ng EEQ kg~(-1) soil(d.w.),其中東北、北方以及東南部分地區(qū)的土壤樣品類雌激素活性較高,然而只有16%的土壤樣品有抗雌激素活性,它莫西芬當量(ER拮抗劑,TEQ)的范圍為0～1535.71μgTEQ kg~(-1) soil(d.w.),平均值為28.37μg FEQ kg~(-1) soil(d.w.),貴州、山西、西藏等地的土壤樣品抗雌激素活性顯著較高。(2)30%土壤提取物樣品具有類雄激素活性,二氫睪酮當量(DEQ)的范圍為0~(-1)05.68 ng DEQ kg~(-1) soil(d.w.),平均值為6.88 ng EEQ kg~(-1) soil(d.w.),山東、浙江、云南等地區(qū)的土壤類雄激素活性較高。54%土壤樣品表現(xiàn)出抗雄激素活性,氟他胺當量(AR拮抗劑,FEQ)范圍為0～410.17μg FEQ kg~(-1) soil(d.w.),平均活性是141.01μg FEQ kg~(-1) soil(d.w.),在中部、東南沿海和西南一些地方的土壤樣品具有較高的抗雄激素活性。(3)32%土壤提取物樣品具有類孕激素活性,孕酮當量(PEQ)的范圍為0-2070.28 ng PEQ kg~(-1) soil(d.w.),平均值為43.10ng PEQ kg~(-1) soil(d.w.),其中福建漳州的土壤提取物表現(xiàn)出非常高的類孕激素活性。73%土壤提取物樣品具有抗孕激素活性,米非司酮當量(PR拮抗劑,REQ)范圍為0～12.58μg REQ kg~(-1) soil(d.w.),平均抗孕激素活性值為2.65μg REQ kg~(-1) soil(d.w.),除遼寧、天津、新疆外,全國的土壤提取物樣品普遍存在抗孕激素活性。(4)35%土壤提取物樣品具有類糖皮質(zhì)激素活性,皮質(zhì)醇當量(HEQ)范圍為0~(-1)48.15 ng HEQ kg~(-1) soil(d.w.),平均值為18.31 ng HEQ kg~(-1) soil(d.w.),上海、江蘇、遼寧等地的土壤樣品類糖皮質(zhì)激素效應(yīng)較顯著;59%土壤提取物樣品具有抗糖皮質(zhì)激素活性,米非司酮當量(GR拮抗劑,REQ)范圍在0～12.26μg REQ kg~(-1) soil(d.w.)之間,平均抗糖皮質(zhì)激素活性值為3.23μg REQ kg~(-1) soil(d.w.),南方地區(qū)土壤樣品的抗糖皮質(zhì)激素活性較北方高,東部較西部高。(5)72%和78%的土壤提取物樣品分別具有類鹽皮質(zhì)激素活性和抗鹽皮質(zhì)激素活性,其中醛固酮當量(AEQ)范圍為0～25.64 ng AEQ kg~(-1) soil(d.w.),平均類鹽皮質(zhì)激素活性為1.82 ng AEQ kg~(-1) soil(d.w.),螺內(nèi)脂當量(MR拮抗劑,SEQ)范圍為0～87.7μg SEQ kg~(-1)(d.w.),平均抗鹽皮質(zhì)激素活性值為27.82μg SEQ kg~(-1)(d.w.),全國各地的土壤樣品廣泛存在類/抗鹽皮質(zhì)激素活性。綜上所述,本研究發(fā)展了以離體細胞實驗的內(nèi)分泌干擾效應(yīng)來評價區(qū)域生態(tài)風險的方法體系,并應(yīng)用這些方法評估了全國農(nóng)田土壤EDCs污染的生態(tài)風險。
[Abstract]:Soil pollution China is more and more serious trend. China's current screening and identification of soil pollutants mainly existing soil environmental quality standard and environmental safety based on the value, but with the rapid development of industry and agriculture, pollutants show variety, wide source characteristics, the existing relevant provisions and standards of basic pollutants according to the Limited toxicity data or lack of effective and accurate detection method of chemical pollutants lack, therefore has been gradually unable to meet the needs of management and control for the modern environmental pollution is difficult. The previous research on soil pollution pollutants focused on the spatial and temporal distribution of ecotoxicological effects due to the complex pollution concern is not high however. The ecological impact of pollutants in the environment and health risks are often long-term low dose exposure results, simply by measuring the concentration of pollutants. The price of its harm and influence is not accurate. Therefore, it is necessary to adopt the method of comprehensive evaluation of soil biological pollution. Our previous study showed that many typical environmental endocrine disruptors (Endocrine Chinese in detection of soil Disrupting Chemicals, EDCs), such as organochlorine pesticides, polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs) and the adjacent benzene two formic acid ester (PAEs). These EDCs can be combined through the simulation of endogenous steroid hormone receptor, produce hormones or hormone activity. The activity of anti steroid hormone receptors including estrogen receptor (Estrogen, Receptor, ER, androgen receptor (Androgen) Receptor, AR (Progesterone), progesterone receptor Receptor. PR), glucocorticoid receptor (Glucocorticoid, Receptor, GR) and mineralocorticoid receptor (Mineralocorticoid, Receptor, MR). There are a large number of studies show that EDCs and ER, AR However, about EDCs and PR, GR, the function of MR is relatively small. There are still many unknown contaminants in the soil environment, the endocrine disrupting effects of biological evaluation methods of soil extract, comprehensive evaluation contains unknown pollutants the endocrine disrupting effects of the overall situation, and therefore more conducive to understanding the ecological risk of EDCs in soil. Study on the acquisition of the country's 31 provinces or regions of 122 topsoil, the extraction of organic pollutants in soil by Soxhlet method, using dual luciferase reporter assay system was used to extract soil by ER, AR, PR, GR and MR mediated / anti steroid hormone activity, the EDCs distribution characteristics and ecological the risk in China's farmland soil. The results of this study show that: (1) 79% soil extract samples the estrogenic activity, estradiol equivalent (EEQ) in the range of 0 ~ 7.34 ng EEQ kg~ (-1) soil (D.W.), 騫沖潎鍊間負1.94 ng EEQ kg~(-1) soil(d.w.),鍏朵腑涓滃寳,鍖楁柟浠ュ強涓滃崡閮ㄥ垎鍦板尯鐨勫湡澹ゆ牱鍝佺被闆屾縺绱犳椿鎬ц緝楂，

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