多溴聯(lián)苯醚的生殖發(fā)育毒性研究
發(fā)布時間：2018-09-04 10:51
【摘要】：多溴聯(lián)苯醚(Polybrominated Diphenyl Ethers, PBDEs)是新型持久性有機污染物。自Main KM等2007年首次報道隱睪癥患兒的患病率與母乳中PBDEs濃度存在關聯(lián)后,針對PBDEs的生殖毒性成為眾多學者關注的研究熱點。 目前,關于PBDEs暴露與人類生殖發(fā)育之間關系的人群研究資料非常有限。有關PBDEs生殖毒性的動物試驗雖然不斷增多,但針對青春期暴露于PBDEs對雄性生殖系統(tǒng)影響的系統(tǒng)報道較少,且所用染毒劑量遠高于實際生活中人群的暴露水平,缺乏對應于人群暴露水平的低劑量毒性研究資料。 針對上述研究不足,本研究首先通過山東某濱海開發(fā)區(qū)建立的出生隊列,就新生兒PBDEs暴露水平及其影響開展前瞻性研究；然后以人群暴露水平作為劑量參考,建立低劑量染毒動物模型,系統(tǒng)觀察青春期前后低劑量BDE-209暴露對雄性生殖系統(tǒng)發(fā)育,特別是精子發(fā)育的影響；最后通過建立睪丸生殖細胞體外培養(yǎng)體系,探索PBDEs影響生殖健康的可能機制。 第一部分某地新生兒多溴聯(lián)苯醚暴露水平調查及其生殖發(fā)育毒性的研究 目的：探討新生兒體內PBDEs暴露水平,分析其暴露與新生兒宮內妊娠結局的相關性。 方法：選擇山東省某濱海開發(fā)區(qū)孕婦作為研究對象,追蹤隨訪孕婦妊娠過程及妊娠結局,同時對新生兒臍帶血血清PBDEs水平進行測定,并對兩者關系進行相關分析。 結果： 新生兒所有血清樣本(n=176)均能檢測到一種或多種PBDEs同系物。新生兒臍帶血清BDE-100(r=0.607, P0.01)、BDE-47(r=0.352, P0.05)、 BDE-99(r=0.412,P0.05)水平與其母親既往懷孕期間因胎兒不正常選擇人工流產呈現(xiàn)明顯的正相關。新生兒血清BDE-153水平與新生兒身長呈顯著正相關(r=0.452,P0.01)。 結論：山東某濱海開發(fā)區(qū)新生兒體內普遍存在PBDEs暴露,新生兒體內PBDEs水平與其宮內不良妊娠發(fā)育結局存在一定的關聯(lián)。 第二部分雄性大鼠青春期前后十溴聯(lián)苯醚暴露的生殖發(fā)育毒性研究 目的：探討青春期前后BDE-209暴露對雄性大鼠性發(fā)育、生殖器腺發(fā)育、精子發(fā)育及生殖內分泌功能的影響。 方法：24只SD雄性大鼠(4周齡)隨機分為4組,以不同濃度BDE-209(0、2、10、50mg/kg.d)灌胃染毒,1次/d,連續(xù)染毒一個月。染毒結束后,采集樣本并分析。 結果： 1、與對照組相比,50mg/kg BDE-209染毒組睪丸抗氧化酶SOD、GSH-Px以及標記酶AKP活性均較對照組明顯下降(P0.05),睪丸病理切片可見空泡樣變。但各染毒組在體重、肛殖距、陰莖包皮分離時間以及各臟器系數(shù)等方面未見明顯改變。 2、BDE-209染毒后,精子活力呈現(xiàn)出隨BDE-209濃度增高而明顯下降的趨勢(P0.05),其中50mg/kg組精子活力下降最為明顯(P0.05)。BDE-209染毒組精子運動速率、前進運動精子率較對照組也呈現(xiàn)出下降趨勢,其中50mg/kg組精子運動速率下降具有統(tǒng)計學意義(P0.05)。與此同時,BDE-209染毒組的精子畸形率較對照組明顯增高(P0.05)。但未見BDE-209暴露對精子密度產生顯著性改變。 3、BDE-209染毒后,血清睪酮、血清雌二醇、睪丸組織睪酮含量與對照組相比,均未見顯著性改變(P0.05)。 結論：在本實驗設計劑量下,青春期前后BDE-209暴露對大鼠睪丸的形態(tài)功能已產生一定的改變,尤其是對精子分化成熟過程產生了明顯影響,引起精子畸形率增加,精子活力下降,精子運動速率降低。 第三部分體外培養(yǎng)睪丸支持細胞暴露于十溴聯(lián)苯醚的毒性研究 目的：探討B(tài)DE-209對睪丸支持細胞的毒性作用。 方法：選用18天齡SD大鼠,取睪丸支持細胞進行原代培養(yǎng),給予BDE-209染毒24h,采用MTT法測定細胞濃度,同時利用Annexin V-FITC試劑盒檢測支持細胞的凋亡情況。 結果： 1、大鼠睪丸支持細胞在培養(yǎng)36h后進入對數(shù)增長期,48-72h是適宜的染毒窗口期。不同濃度BDE-209染毒24h后,隨染毒濃度增加,細胞存活率下降,其中當BDE-209濃度≥30μg/ml時,睪丸支持細胞的相對存活率較對照組明顯降低(P0.05)。 2、與對照組相比,BDE-209染毒組支持細胞T-AOC、SOD呈現(xiàn)下降趨勢,其中10、20μg/ml染毒組T-AOC、SOD活性較對照組明顯下降(P0.05)。 3、BDE-209染毒24h后,睪丸支持細胞的凋亡率出現(xiàn)增高趨勢,當BDE-209濃度≥20μg/ml時,睪丸支持細胞凋亡率較對照組顯著增高(P0.05)。 結論：BDE-209對體外培養(yǎng)的睪丸支持細胞具有細胞毒性作用,BDE-209暴露可引起睪丸支持細胞出現(xiàn)凋亡。
[Abstract]:Polybrominated Diphenyl Ethers (PBDEs) is a new type of persistent organic pollutant. Since the incidence of cryptorchidism in children was first reported by Main KM in 2007, the reproductive toxicity of PBDEs in breast milk has become a research hotspot.
At present, there is very limited data on the relationship between PBDEs exposure and human reproductive development. Animal trials on the reproductive toxicity of PBDEs are increasing, but systematic reports on the effects of pubertal exposure to PBDEs on the male reproductive system are scarce, and the dose used is much higher than the exposure level in real life. Lack of data for low dose toxicity studies corresponding to population exposure levels.
In view of the insufficiency of the above studies, this study firstly carried out a prospective study on the level of PBDEs exposure and its effects in newborns through a birth cohort in a coastal development area of Shandong Province, and then established an animal model of low-dose exposure to BDE-209 before and after puberty, taking the level of population exposure as a dose reference, to systematically observe the effects of low-dose BDE-209 exposure on male puberty. The effects of PBDEs on reproductive system development, especially sperm development, and the possible mechanism of PBDEs on reproductive health were explored by establishing an in vitro culture system of testicular germ cells.
The first part is a survey of the exposure level of polybrominated diphenyl ethers in a newborn and its reproductive toxicity.
Objective: To investigate the level of PBDEs exposure in neonates and analyze the correlation between PBDEs exposure and intrauterine pregnancy outcome.
Methods: The pregnant women in a coastal development area of Shandong Province were selected as the research object. The pregnancy process and pregnancy outcome were followed up. The serum PBDEs levels of umbilical cord blood were measured and the relationship between them was analyzed.
Result:
One or more PBDEs homologues were detected in all neonatal serum samples (n = 176). The levels of BDE-100 (r = 0.607, P 0.01), BDE-47 (r = 0.352, P 0.05), BDE-99 (r = 0.412, P 0.05) in neonatal umbilical cord serum were positively correlated with maternal abnormal selection of induced abortion during previous pregnancy. Neonatal length was positively correlated (r=0.452, P0.01).
Conclusion: PBDEs are ubiquitous in newborns of a coastal development zone in Shandong Province. The level of PBDEs in newborns is related to the adverse pregnancy outcomes.
The second part is about the reproductive and developmental toxicity of ten brominated diphenyl ethers before and after puberty in male rats.
Objective: To investigate the effects of BDE-209 exposure on sexual development, genital gland development, sperm development and reproductive endocrine function in male rats before and after puberty.
METHODS: Twenty-four SD male rats (4 weeks old) were randomly divided into four groups. The rats were given different concentrations of BDE-209 (0,2,10,50 mg/kg.d) by gastric Administration for one month.
Result:
1. Compared with the control group, the activities of SOD, GSH-Px and AKP in testis of 50 mg/kg BDE-209 poisoning group were significantly lower than those of the control group (P 0.05). The pathological section of testis showed vacuole-like changes.
2. After BDE-209 exposure, the sperm motility decreased significantly with the increase of BDE-209 concentration (P 0.05), especially in the 50mg/kg group (P 0.05). Meanwhile, the sperm deformity rate of BDE-209 exposed group was significantly higher than that of the control group (P 0.05). However, there was no significant change in sperm density after exposure to BDE-209.
3. There were no significant changes in serum testosterone, serum estradiol and testosterone content in testicular tissue between BDE-209 and control group (P 0.05).
CONCLUSION: BDE-209 exposure before and after puberty has a certain effect on the morphology and function of rat testis, especially on the process of sperm differentiation and maturation, resulting in an increase in the rate of sperm deformity, a decrease in sperm motility and a decrease in sperm motility.
The third part is the study of the toxicity of Sertoli cells exposed to ten brominated diphenyl ethers in vitro.
Objective: To investigate the toxic effects of BDE-209 on Sertoli cells.
METHODS: Sertoli cells from 18-day-old SD rats were cultured for 24 hours after exposure to BDE-209. MTT assay was used to determine the concentration of Sertoli cells and Annexin V-FITC kit was used to detect the apoptosis of Sertoli cells.
Result:
1. Rat Sertoli cells entered the logarithmic growth phase after 36 hours of culture, and 48-72 hours was the appropriate exposure window period. After 24 hours of exposure to different concentrations of BDE-209, the cell survival rate decreased with the increase of the concentration of BDE-209. The relative survival rate of Sertoli cells decreased significantly when the concentration of BDE-209 was greater than 30 ug/ml (P 0.05).
2. Compared with the control group, the activity of T-AOC and SOD in Sertoli cells of BDE-209 exposed group showed a downward trend. The activity of T-AOC and SOD in 10,20 ug/ml exposed group was significantly lower than that in the control group (P 0.05).
3. After 24 hours of BDE-209 exposure, the apoptotic rate of Sertoli cells increased. When the concentration of BDE-209 was more than 20 ug/ml, the apoptotic rate of Sertoli cells increased significantly (P 0.05).
Conclusion: BDE-209 has cytotoxic effect on Sertoli cells cultured in vitro, and BDE-209 exposure can induce apoptosis of Sertoli cells.
【學位授予單位】：華中科技大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R114
本文編號：2221862
[Abstract]:Polybrominated Diphenyl Ethers (PBDEs) is a new type of persistent organic pollutant. Since the incidence of cryptorchidism in children was first reported by Main KM in 2007, the reproductive toxicity of PBDEs in breast milk has become a research hotspot.
At present, there is very limited data on the relationship between PBDEs exposure and human reproductive development. Animal trials on the reproductive toxicity of PBDEs are increasing, but systematic reports on the effects of pubertal exposure to PBDEs on the male reproductive system are scarce, and the dose used is much higher than the exposure level in real life. Lack of data for low dose toxicity studies corresponding to population exposure levels.
In view of the insufficiency of the above studies, this study firstly carried out a prospective study on the level of PBDEs exposure and its effects in newborns through a birth cohort in a coastal development area of Shandong Province, and then established an animal model of low-dose exposure to BDE-209 before and after puberty, taking the level of population exposure as a dose reference, to systematically observe the effects of low-dose BDE-209 exposure on male puberty. The effects of PBDEs on reproductive system development, especially sperm development, and the possible mechanism of PBDEs on reproductive health were explored by establishing an in vitro culture system of testicular germ cells.
The first part is a survey of the exposure level of polybrominated diphenyl ethers in a newborn and its reproductive toxicity.
Objective: To investigate the level of PBDEs exposure in neonates and analyze the correlation between PBDEs exposure and intrauterine pregnancy outcome.
Methods: The pregnant women in a coastal development area of Shandong Province were selected as the research object. The pregnancy process and pregnancy outcome were followed up. The serum PBDEs levels of umbilical cord blood were measured and the relationship between them was analyzed.
Result:
One or more PBDEs homologues were detected in all neonatal serum samples (n = 176). The levels of BDE-100 (r = 0.607, P 0.01), BDE-47 (r = 0.352, P 0.05), BDE-99 (r = 0.412, P 0.05) in neonatal umbilical cord serum were positively correlated with maternal abnormal selection of induced abortion during previous pregnancy. Neonatal length was positively correlated (r=0.452, P0.01).
Conclusion: PBDEs are ubiquitous in newborns of a coastal development zone in Shandong Province. The level of PBDEs in newborns is related to the adverse pregnancy outcomes.
The second part is about the reproductive and developmental toxicity of ten brominated diphenyl ethers before and after puberty in male rats.
Objective: To investigate the effects of BDE-209 exposure on sexual development, genital gland development, sperm development and reproductive endocrine function in male rats before and after puberty.
METHODS: Twenty-four SD male rats (4 weeks old) were randomly divided into four groups. The rats were given different concentrations of BDE-209 (0,2,10,50 mg/kg.d) by gastric Administration for one month.
Result:
1. Compared with the control group, the activities of SOD, GSH-Px and AKP in testis of 50 mg/kg BDE-209 poisoning group were significantly lower than those of the control group (P 0.05). The pathological section of testis showed vacuole-like changes.
2. After BDE-209 exposure, the sperm motility decreased significantly with the increase of BDE-209 concentration (P 0.05), especially in the 50mg/kg group (P 0.05). Meanwhile, the sperm deformity rate of BDE-209 exposed group was significantly higher than that of the control group (P 0.05). However, there was no significant change in sperm density after exposure to BDE-209.
3. There were no significant changes in serum testosterone, serum estradiol and testosterone content in testicular tissue between BDE-209 and control group (P 0.05).
CONCLUSION: BDE-209 exposure before and after puberty has a certain effect on the morphology and function of rat testis, especially on the process of sperm differentiation and maturation, resulting in an increase in the rate of sperm deformity, a decrease in sperm motility and a decrease in sperm motility.
The third part is the study of the toxicity of Sertoli cells exposed to ten brominated diphenyl ethers in vitro.
Objective: To investigate the toxic effects of BDE-209 on Sertoli cells.
METHODS: Sertoli cells from 18-day-old SD rats were cultured for 24 hours after exposure to BDE-209. MTT assay was used to determine the concentration of Sertoli cells and Annexin V-FITC kit was used to detect the apoptosis of Sertoli cells.
Result:
1. Rat Sertoli cells entered the logarithmic growth phase after 36 hours of culture, and 48-72 hours was the appropriate exposure window period. After 24 hours of exposure to different concentrations of BDE-209, the cell survival rate decreased with the increase of the concentration of BDE-209. The relative survival rate of Sertoli cells decreased significantly when the concentration of BDE-209 was greater than 30 ug/ml (P 0.05).
2. Compared with the control group, the activity of T-AOC and SOD in Sertoli cells of BDE-209 exposed group showed a downward trend. The activity of T-AOC and SOD in 10,20 ug/ml exposed group was significantly lower than that in the control group (P 0.05).
3. After 24 hours of BDE-209 exposure, the apoptotic rate of Sertoli cells increased. When the concentration of BDE-209 was more than 20 ug/ml, the apoptotic rate of Sertoli cells increased significantly (P 0.05).
Conclusion: BDE-209 has cytotoxic effect on Sertoli cells cultured in vitro, and BDE-209 exposure can induce apoptosis of Sertoli cells.
【學位授予單位】：華中科技大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R114
【參考文獻】
相關期刊論文 前10條
1 張作濤;謝高宇;陳凱;齊敏友;劉浩然;;腺嘌呤致大鼠睪丸損傷中標志酶的變化及淫羊藿總黃酮的干預作用[J];重慶醫(yī)學;2012年07期
2 王志新;段華英;王玲;陳敦金;;外源性BDE-209對小鼠精子DNA的損傷[J];南方醫(yī)科大學學報;2011年01期
3 羅賢麗;劉麗華;晉藝聰;;多溴聯(lián)苯醚動物毒性綜述[J];化學工程與裝備;2010年01期
4 齊彭德;陸光華;梁艷;趙海洲;;多溴聯(lián)苯醚的生物效應研究[J];環(huán)境科學與技術;2011年11期
5 任金亮;王平;;多溴聯(lián)苯醚環(huán)境行為的特征與研究進展[J];化工進展;2006年10期
6 ;Polybrominated diphenyl ethers in atmosphere and soil of a production area in China:Levels and partitioning[J];Journal of Environmental Sciences;2011年03期
7 何衛(wèi)紅;何平;張明;夏濤;陳學敏;王愛國;;2,2’,4,4’-四溴聯(lián)苯醚對原代新生大鼠海馬細胞氧化應激、DNA損傷和凋亡的影響[J];環(huán)境與健康雜志;2007年01期
8 于功昌;王筱芬;劉衍忠;謝琳;李莉;;苯并咪唑對大鼠生精功能和睪丸酶活力的影響[J];環(huán)境與健康雜志;2008年08期
9 梁淑軒;李立勇;張國峰;姜靜;高紅霞;趙燕燕;張建生;;十溴聯(lián)苯醚在小鼠體內的富集代謝[J];環(huán)境與健康雜志;2010年04期
10 李祥;湯艷;熊偉;于雪嵐;劉揚;;BDE-209對雄性大鼠睪丸組織結構和氧化應激的影響[J];環(huán)境與健康雜志;2011年12期
相關博士學位論文 前1條
1 于珊珊;DNA氧化損傷修復反應體系及其對細胞壽命影響機制初探[D];山東大學;2009年
，本文編號：2221862
本文鏈接：http://sikaile.net/yixuelunwen/yufangyixuelunwen/2221862.html
最近更新
教材專著
