內分泌干擾物雙酚A對秀麗隱桿線蟲的生態(tài)毒性效應及其作用機制研究
發(fā)布時間：2018-11-08 15:57
【摘要】：雙酚A(BPA)是世界公認的環(huán)境內分泌干擾物,一直是毒理學研究中的明星化合物。BPA的世界年需求量超過400萬噸/年,環(huán)境中的污染也日趨嚴重,在土壤、水、大氣和沉積物中都發(fā)現(xiàn)了BPA的蹤跡,甚至在人體組織和體液中也檢測到了BPA的存在。體內和體外實驗已經證明BPA暴露能夠引起內分泌干擾效應、神經毒性、生殖毒性、免疫毒性和致癌致畸性等。人群流行病學研究也發(fā)現(xiàn),糖尿病、肥胖、神經行為障礙、肝功能異常、復發(fā)性流產、發(fā)育性疾病都與BPA暴露有關。目前關于BPA暴露對無脊椎動物的毒性報道較少。作為一種從土壤中分離出來的代表性的線蟲種類,秀麗隱桿線蟲是環(huán)境暴露評價以及生態(tài)毒理學研究中最常用的模式生物之一。本研究選取秀麗隱桿線蟲作為受試生物,主要從生理、生化、分子水平探究環(huán)境相關濃度下BPA急性與亞急性暴露、慢性暴露和多代連續(xù)暴露對秀麗隱桿線蟲的生態(tài)毒性效應；此外,本研究還考察了BPA的替代品雙酚S(BPS)對秀麗隱桿線蟲的毒性影響,并與BPA的毒性進行對比,分析BPA替代品的生態(tài)風險性。本研究主要的研究結果如下：(1)BPA急性與亞急性暴露能夠顯著抑制秀麗隱桿線蟲的體長、運動行為水平以及子代數目,顯著提高細胞凋亡水平以及脅迫相關基因的表達量。急性暴露后,秀麗隱桿線蟲的頭部擺動頻率是最敏感的生理指標,暴露濃度在0.01μM即可對秀麗隱桿線蟲產生顯著的負面效應,而亞急性暴露后,體長和頭部擺動頻率相比其他生理指標更加敏感,同樣在暴露濃度為0.01μM時能夠對秀麗隱桿線蟲產生顯著的抑制作用。BPA急性和亞急性暴露對線蟲體內脂褐素累積和ROS水平沒有產生顯著影響,但當暴露濃度達到0.1μM時能夠顯著提高線蟲體內細胞凋亡水平,表明實驗濃度下BPA暴露對秀麗隱桿線蟲產生的毒性效應并不是通過氧化損傷途徑,而很可能與細胞凋亡水平升高有關。BPA亞急性暴露能夠導致秀麗隱桿線蟲體內絕大部分脅迫相關基因表達量的升高,即使在最低暴露濃度0.001 μM, BPA暴露也能夠引起絕大多數脅迫相關基因表達量的顯著提升。(2)BPA慢性暴露能夠顯著抑制秀麗隱桿線蟲的體長、運動行為水平、壽命以及種群數目,顯著提高線蟲體內細胞凋亡水平、脅迫相關基因表達以及轉基因品系TJ375的熒光蛋白表達量。慢性暴露后,頭部擺動頻率是最敏感的生理指標,暴露濃度在0.001μM即可對秀麗隱桿線蟲產生顯著的負面效應。BPA慢性暴露對線蟲體內脂褐素累積同樣沒有顯著影響,但能夠在暴露濃度達到0.01μM以上時,顯著地提高秀麗隱桿線蟲體內的細胞凋亡水平。在實驗濃度范圍內,BPA慢性暴露能夠明顯地提高線蟲體內絕大部分脅迫相關基因的表達量。與亞急性暴露的結果相比,同一暴毒濃度下,BPA慢性暴露能夠引起脅迫相關基因的mRNA水平更為明顯的提高。當BPA暴露濃度達到0.1μM時,秀麗隱桿線蟲的種群數目顯著減小。轉基因品系TJ375只能夠檢測環(huán)境濃度達到1μM時的BPA毒性,與其它測試指標相比不夠靈敏。(3)BPA多代連續(xù)暴露能夠在暴露濃度達到0.01μM以上時對秀麗隱桿線蟲的生理指標產生顯著的毒性影響。在較低暴露濃度時,子代線蟲會出現(xiàn)毒性逐代修復或者惡化的情況,造成秀麗隱桿線蟲生理水平上的世代毒性差異,而在最高暴露濃度10μM時,秀麗隱桿線蟲G1代和G4代在毒性效應上沒有顯著性的差異。在秀麗隱桿線蟲G1代和G4代中,與高濃度暴露組相比,低濃度(≤0.01 μM)暴露組能夠引起體內脅迫相關基因表達量更為明顯的提高；與G4代相比,秀麗隱桿線蟲的G1代體內脅迫相關基因的表達量增加更為顯著。(4)BPS亞急性暴露后,秀麗隱桿線蟲的頭部擺動頻率是最敏感的生理指標,暴露濃度在1 μM以上才能夠對秀麗隱桿線蟲產生顯著的負面效應,而其他生理指標只有在最高濃度暴露組才會受到顯著的負面影響。亞急性暴露后,秀麗隱桿線蟲體內脂褐素的累積沒有受到顯著影響,細胞凋亡水平也只有在暴露濃度為1μM時顯著提高。BPS亞急性暴露能夠引起秀麗隱桿線蟲體內絕大部分基因表達量的小幅度上升。綜上所述,低濃度的BPA暴露能夠對秀麗隱桿線蟲產生發(fā)育毒性、神經行為毒性以及生殖毒性。BPA暴露對秀麗隱桿線蟲生理水平的毒性影響很可能與體內細胞凋亡水平提高有關；BPA暴露能夠顯著提高秀麗隱桿線蟲體內脅迫相關基因的表達,破壞機體的穩(wěn)態(tài)；轉基因品系TJ375對于BPA暴露不敏感,不能有效監(jiān)測BPA的毒性；與BPA毒性相比,BPS的毒性明顯較弱。
[Abstract]:Bisphenol A (BPA) is a world-recognized environmental endocrine disruptor and has been a star compound in toxicology studies. The world's annual demand for BPA is more than 4 million tons/ year, and the pollution in the environment is becoming more and more serious. The trace of BPA is found in soil, water, atmosphere and sediment, and BPA is also detected in human tissue and body fluids. In vivo and in vitro experiments have shown that the exposure of BPA can cause endocrine disruption, neurotoxicity, reproductive toxicity, immunotoxicity, and carcinogenicity. The epidemiological study of the population also found that diabetes, obesity, neurobehavioral disorders, liver function abnormalities, recurrent spontaneous abortion, and developmental disorders were associated with exposure to BPA. At present, the toxicity of BPA exposure to invertebrates is less. As a representative nematode species isolated from the soil, the Caenorhabditis elegans is one of the most commonly used modes in environmental exposure assessment and ecological toxicology studies. The study was conducted to study the ecological toxicity of Caenorhabditis elegans as the tested organism, mainly from the physiological, biochemical and molecular level to the acute and subacute exposure of BPA, the chronic exposure and the continuous exposure of multiple generations to the enorhabditis elegans; in addition, The toxicity of biphenol S (BPS) to the caenorhabditis elegans was also investigated in this study, and the toxicity of BPA was compared with the toxicity of BPA, and the ecological risk of the substitute of BPA was analyzed. The main results of this study are as follows: (1) The acute and subacute exposure of BPA can significantly inhibit the body length, the level of activity and the number of offspring of the caenorhabditis elegans, and significantly increase the level of cell apoptosis and the expression of stress-related genes. after acute exposure, the head swing frequency of the caenorhabditis elegans is the most sensitive physiological index, and the exposure concentration is 0. 01 & mu; m, which can produce a significant negative effect on the caenorhabditis elegans, and after the subacute exposure, the body length and the head swing frequency are more sensitive than other physiological indexes, It is also possible to have a significant inhibitory effect on the caenorhabditis elegans at a concentration of 0.01. m u.M. The acute and subacute exposure of BPA did not have a significant effect on the accumulation of lipofuscin and the level of ROS in the nematode, but when the exposure concentration reached 0.1. m The results show that the toxic effect of BPA exposure to the caenorhabditis elegans is not the way of oxidative damage, and it is likely to be related to the increase of the level of apoptosis. The subacute exposure of BPA can lead to a significant increase in the expression of most stress-related genes in the Caenorhabditis elegans, even when the minimum exposure concentration is 0. 001. m u.M, and the exposure of BPA can also cause a significant increase in the expression of most stress-related genes. (2) The chronic exposure of BPA can significantly inhibit the body length, the activity level, the service life and the population number of the caenorhabditis elegans, and obviously improve the cell apoptosis level, the stress-related gene expression and the fluorescent protein expression amount of the transgenic line TJ375. After chronic exposure, the swing frequency of the head is the most sensitive physiological index, and the exposure concentration is 0. 001. m The chronic exposure of BPA did not have a significant effect on the accumulation of lipofuscin in Nematode, but it was able to significantly increase the level of apoptosis in the Caenorhabditis elegans when the exposure concentration was above 0.01. m In the experimental concentration range, the chronic exposure of BPA can significantly improve the expression of most stress-related genes in the nematode. Compared with the results of subacute exposure, the chronic exposure of BPA could lead to a more significant increase in the mRNA level of stress-related genes. When the exposure concentration of BPA reached 0.1. m u.M, the number of Caenorhabditis elegans was significantly reduced. The transgenic strain TJ375 can only detect the toxicity of BPA when the environmental concentration reaches 1. m u.M, and is not sensitive to other test indicators. (3) The continuous exposure of BPA can have a significant effect on the physiological index of the Caenorhabditis elegans when the exposure concentration is above 0.01. m In the case of lower exposure, the generation toxicity of Nematode in the offspring was different from generation to generation, while at the highest exposure concentration of 10. m u.M, there was no significant difference in the toxicity effect of the G1 and G4 of the Caenorhabditis elegans. in that G1 and G4 generation of the Caenorhabditis elegans, the exposure group of low concentration (0. 01. mu.M) can induce a more significant increase in the expression of the related gene in the in vivo compared with the high-concentration exposure group; compared with the G4 generation, The expression of the stress-related genes in the G1-generation of the Caenorhabditis elegans was more significant. (4) After the subacute exposure of BPS, the head swing frequency of the caenorhabditis elegans is the most sensitive physiological index, and the exposure concentration is more than 1. m while other physiological indicators will be significantly negatively affected only in the highest concentration exposure group. After sub-acute exposure, the accumulation of lipofuscin in the caenorhabditis elegans was not significantly affected, and the level of apoptosis was significantly increased only when the exposure concentration was 1. m The sub-acute exposure of BPS can cause a small increase in the expression of most of the gene in the Caenorhabditis elegans. In conclusion, exposure of low concentrations of BPA can produce developmental toxicity, neurobehavioral toxicity, and reproductive toxicity to the Caenorhabditis elegans. The effect of BPA exposure on the physiological level of Caenorhabditis elegans is likely to be related to the improvement of the level of cell apoptosis in vivo; the exposure of BPA can significantly improve the expression of stress-related genes in the Caenorhabditis elegans and destroy the steady state of the body; the transgenic line TJ375 is not sensitive to the exposure of BPA, The toxicity of BPA could not be effectively monitored; the toxicity of BPS was significantly less than the toxicity of BPA.
【學位授予單位】：華東理工大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：X171.5
本文編號：2318956
[Abstract]:Bisphenol A (BPA) is a world-recognized environmental endocrine disruptor and has been a star compound in toxicology studies. The world's annual demand for BPA is more than 4 million tons/ year, and the pollution in the environment is becoming more and more serious. The trace of BPA is found in soil, water, atmosphere and sediment, and BPA is also detected in human tissue and body fluids. In vivo and in vitro experiments have shown that the exposure of BPA can cause endocrine disruption, neurotoxicity, reproductive toxicity, immunotoxicity, and carcinogenicity. The epidemiological study of the population also found that diabetes, obesity, neurobehavioral disorders, liver function abnormalities, recurrent spontaneous abortion, and developmental disorders were associated with exposure to BPA. At present, the toxicity of BPA exposure to invertebrates is less. As a representative nematode species isolated from the soil, the Caenorhabditis elegans is one of the most commonly used modes in environmental exposure assessment and ecological toxicology studies. The study was conducted to study the ecological toxicity of Caenorhabditis elegans as the tested organism, mainly from the physiological, biochemical and molecular level to the acute and subacute exposure of BPA, the chronic exposure and the continuous exposure of multiple generations to the enorhabditis elegans; in addition, The toxicity of biphenol S (BPS) to the caenorhabditis elegans was also investigated in this study, and the toxicity of BPA was compared with the toxicity of BPA, and the ecological risk of the substitute of BPA was analyzed. The main results of this study are as follows: (1) The acute and subacute exposure of BPA can significantly inhibit the body length, the level of activity and the number of offspring of the caenorhabditis elegans, and significantly increase the level of cell apoptosis and the expression of stress-related genes. after acute exposure, the head swing frequency of the caenorhabditis elegans is the most sensitive physiological index, and the exposure concentration is 0. 01 & mu; m, which can produce a significant negative effect on the caenorhabditis elegans, and after the subacute exposure, the body length and the head swing frequency are more sensitive than other physiological indexes, It is also possible to have a significant inhibitory effect on the caenorhabditis elegans at a concentration of 0.01. m u.M. The acute and subacute exposure of BPA did not have a significant effect on the accumulation of lipofuscin and the level of ROS in the nematode, but when the exposure concentration reached 0.1. m The results show that the toxic effect of BPA exposure to the caenorhabditis elegans is not the way of oxidative damage, and it is likely to be related to the increase of the level of apoptosis. The subacute exposure of BPA can lead to a significant increase in the expression of most stress-related genes in the Caenorhabditis elegans, even when the minimum exposure concentration is 0. 001. m u.M, and the exposure of BPA can also cause a significant increase in the expression of most stress-related genes. (2) The chronic exposure of BPA can significantly inhibit the body length, the activity level, the service life and the population number of the caenorhabditis elegans, and obviously improve the cell apoptosis level, the stress-related gene expression and the fluorescent protein expression amount of the transgenic line TJ375. After chronic exposure, the swing frequency of the head is the most sensitive physiological index, and the exposure concentration is 0. 001. m The chronic exposure of BPA did not have a significant effect on the accumulation of lipofuscin in Nematode, but it was able to significantly increase the level of apoptosis in the Caenorhabditis elegans when the exposure concentration was above 0.01. m In the experimental concentration range, the chronic exposure of BPA can significantly improve the expression of most stress-related genes in the nematode. Compared with the results of subacute exposure, the chronic exposure of BPA could lead to a more significant increase in the mRNA level of stress-related genes. When the exposure concentration of BPA reached 0.1. m u.M, the number of Caenorhabditis elegans was significantly reduced. The transgenic strain TJ375 can only detect the toxicity of BPA when the environmental concentration reaches 1. m u.M, and is not sensitive to other test indicators. (3) The continuous exposure of BPA can have a significant effect on the physiological index of the Caenorhabditis elegans when the exposure concentration is above 0.01. m In the case of lower exposure, the generation toxicity of Nematode in the offspring was different from generation to generation, while at the highest exposure concentration of 10. m u.M, there was no significant difference in the toxicity effect of the G1 and G4 of the Caenorhabditis elegans. in that G1 and G4 generation of the Caenorhabditis elegans, the exposure group of low concentration (0. 01. mu.M) can induce a more significant increase in the expression of the related gene in the in vivo compared with the high-concentration exposure group; compared with the G4 generation, The expression of the stress-related genes in the G1-generation of the Caenorhabditis elegans was more significant. (4) After the subacute exposure of BPS, the head swing frequency of the caenorhabditis elegans is the most sensitive physiological index, and the exposure concentration is more than 1. m while other physiological indicators will be significantly negatively affected only in the highest concentration exposure group. After sub-acute exposure, the accumulation of lipofuscin in the caenorhabditis elegans was not significantly affected, and the level of apoptosis was significantly increased only when the exposure concentration was 1. m The sub-acute exposure of BPS can cause a small increase in the expression of most of the gene in the Caenorhabditis elegans. In conclusion, exposure of low concentrations of BPA can produce developmental toxicity, neurobehavioral toxicity, and reproductive toxicity to the Caenorhabditis elegans. The effect of BPA exposure on the physiological level of Caenorhabditis elegans is likely to be related to the improvement of the level of cell apoptosis in vivo; the exposure of BPA can significantly improve the expression of stress-related genes in the Caenorhabditis elegans and destroy the steady state of the body; the transgenic line TJ375 is not sensitive to the exposure of BPA, The toxicity of BPA could not be effectively monitored; the toxicity of BPS was significantly less than the toxicity of BPA.
【學位授予單位】：華東理工大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：X171.5
【參考文獻】
相關期刊論文 前10條
1 李圓圓;付旭鋒;趙亞嫻;蘇紅巧;秦占芬;;雙酚A與其替代品對黑斑蛙急性毒性的比較[J];生態(tài)毒理學報;2015年02期
2 余建龍;李明;陳紅兵;邵兵;;雙酚S在T47D乳腺癌細胞中雌激素活性的研究[J];毒理學雜志;2014年02期
3 周海東;黃霞;王曉琳;文湘華;;北京市城市污水雌激素活性的研究[J];環(huán)境科學;2009年12期
4 董軍;李向麗;梁銳杰;;珠江口地區(qū)水體中雙酚A污染及其與環(huán)境因子的關系[J];生態(tài)與農村環(huán)境學報;2009年02期
5 ;Using the nematode Caenorhabditis elegans as a model animal for assessing the toxicity induced by microcystin-LR[J];Journal of Environmental Sciences;2009年03期
6 沈露露;王大勇;;秀麗線蟲衰老調控的生理與分子機制[J];生理科學進展;2009年01期
7 邵曉玲;馬軍;文剛;;松花江流域某自來水廠中內分泌干擾物的調查[J];環(huán)境科學;2008年10期
8 ;Assessment of locomotion behavioral defects induced by acute toxicity from heavy metal exposure in nematode Caenorhabditis elegans[J];Journal of Environmental Sciences;2008年09期
9 張燕芬;王大勇;;利用模式動物秀麗線蟲建立環(huán)境毒物毒性的評估研究體系[J];生態(tài)毒理學報;2008年04期
10 雷炳莉;駱堅平;查金苗;黃圣彪;劉操;王子健;;溫榆河沉積物中壬基酚和雙酚A的分布[J];環(huán)境化學;2008年03期
相關博士學位論文 前2條
1 張淑華;萘對秀麗線蟲的毒性作用及DhHP-6的體內解毒效應[D];東北師范大學;2011年
2 田雨;久效磷對秀麗隱桿線蟲（Caenorhabditis elegans）多種毒性效應的當量評價[D];中國海洋大學;2011年
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