本文選題：雙酚A + 自發(fā)性行為�。� 參考：《山西農(nóng)業(yè)大學(xué)》2014年博士論文

【摘要】：[目的]雙酚A (BisphenolA, BPA)作為具有雌激素活性的環(huán)境內(nèi)分泌干擾物,是環(huán)氧樹脂和碳酸乙酯的主要材料,對(duì)發(fā)育中的中樞神經(jīng)系統(tǒng),特別是學(xué)習(xí)記憶有副作用,但BPA影響學(xué)習(xí)記憶的機(jī)制還不是很清楚。本文主要從表觀遺傳學(xué)、信號(hào)通路和突觸可塑性三個(gè)方面探討了胚胎期BPA暴露對(duì)子代雄性大鼠學(xué)習(xí)記憶能力影響的分子機(jī)制,為進(jìn)一步闡明BPA的神經(jīng)毒性機(jī)制奠定理論基礎(chǔ),為今后實(shí)施科學(xué)診斷和靶標(biāo)藥物設(shè)計(jì)提供實(shí)驗(yàn)室依據(jù)。[方法]懷孕的Sprague-Dawley大鼠從懷孕第9天(E9)到懷孕第20天(E 20)口服0.05、0.5、5或50 mg/kg-body weight (BW) per day的BPA,選取出生后21天(PND21)的雄性小鼠進(jìn)行相關(guān)試驗(yàn)。采用開場(chǎng)試驗(yàn)、目標(biāo)識(shí)別試驗(yàn)和八臂迷宮試驗(yàn)檢測(cè)雄性仔鼠的自發(fā)性行為和學(xué)習(xí)記憶能力；采用HE染色、透射電鏡、Real-time PCR, Western blotting等技術(shù)手段,檢測(cè)了雄性仔鼠的腦組織形態(tài)結(jié)構(gòu)、海馬CA1區(qū)超微結(jié)構(gòu)、表觀遺傳相關(guān)酶類(DNMTs和HDACs)、ERK信號(hào)通路相關(guān)分子、突觸標(biāo)記物(synaptophysin, postsynaptic density-95 (PSD-95), spinophilin)、谷氨酸受體(G1uR1和NMDARl) mRNA和蛋白的表達(dá),從而探討胚胎期BPA暴露引起神經(jīng)毒性的分子機(jī)制。[結(jié)果](1)胚胎期BPA暴露影響了出生后雄性仔鼠的生理生化指標(biāo),包括增加了出生后第9天、14天和21天雄性小鼠的體重；降低了腦、海馬和睪丸的臟體比；增加了血清睪酮濃度,降低了血清雌激素水平；同時(shí),誘導(dǎo)了雄性仔鼠海馬組織形態(tài)學(xué)的病理損傷：海馬各區(qū)錐體細(xì)胞和顆粒細(xì)胞明顯體積變小,數(shù)量減少,細(xì)胞間隙增大,神經(jīng)元皺縮,細(xì)胞邊緣不清晰。(2)胚胎期BPA暴露影響了出生后雄性仔鼠在開場(chǎng)試驗(yàn)中的自發(fā)性行為,包括降低了運(yùn)動(dòng)活性、減少了探索行為和理毛行為；損害了目標(biāo)識(shí)別實(shí)驗(yàn)中的短期記憶和長期記憶；增加了八臂迷宮中的參考記憶特別是工作記憶錯(cuò)誤次數(shù)。(3)胚胎期BPA暴露影響了出生后雄性仔鼠海馬中相關(guān)表觀遺傳酶的表達(dá),Real-time PCR結(jié)果顯示BPA暴露抑制了DNA甲基轉(zhuǎn)移酶DNMT1 mRNA和組蛋白去乙�；窰DAC1 mRNA表達(dá)水平,上調(diào)了組蛋白去乙酰化酶HDAC2 mRNA表達(dá)水平。(4)胚胎期BPA暴露影響了出生后雄性仔鼠海馬中ERK通路相關(guān)分子的表達(dá),Western Blotting數(shù)據(jù)顯示,與對(duì)照組相比,BPA暴露組雄性小鼠海馬中Akt、phospho-Akt、 p44/42 MAPK、phospho-p44/42 MAPK、phospho-CREB、BDNF蛋白表達(dá)水平顯著降低。(5)胚胎期BPA暴露影響了出生后雄性仔鼠海馬CA1區(qū)突觸超微結(jié)構(gòu)：包括突觸間隙變寬、PSD區(qū)變薄、突觸表面模糊不清、突觸囊泡消失；Real-time PCR和Western Blotting實(shí)驗(yàn)數(shù)據(jù)表明,BPA暴露下調(diào)了突觸標(biāo)記性分子(synaptophysin, PSD-95,和spinophilin) mRNA和蛋白表達(dá)；抑制了谷氨酸受體(G1uR1和NMDARl) mRNA和蛋白表達(dá)。[結(jié)論]本課題實(shí)驗(yàn)結(jié)果揭示了胚胎期BPA暴露誘導(dǎo)了出生后雄性仔鼠的學(xué)習(xí)記憶能力的降低,其分子機(jī)制涉及到表觀遺傳學(xué)、ERK通路和突觸可塑性的改變,提示BPA影響中樞神經(jīng)系統(tǒng)和行為發(fā)育機(jī)制的復(fù)雜性。
[Abstract]:[Objective] bisphenol A (BisphenolA, BPA), as an environmental endocrine disruptor with estrogen activity, is the main material of epoxy and ethyl carbonate. It has side effects on the developing central nervous system, especially learning and memory, but the mechanism that BPA affects learning and memory is not clear. This article mainly from epigenetics, signal pathway The three aspects of synaptic plasticity discussed the molecular mechanism of the effects of BPA exposure on the learning and memory ability of the offspring of the offspring of the offspring, which lay a theoretical basis for further clarifying the neurotoxic mechanism of BPA, and provided a laboratory basis for the future implementation of scientific diagnosis and target drug design. [methods] the pregnant Sprague-Dawley rats were ninth from pregnancy. Days (E9) to twentieth days of pregnancy (E 20) oral 0.05,0.5,5 or BPA of 50 mg/kg-body weight (BW) per day, selected male mice of 21 days after birth (PND21). The spontaneous behavior and learning and memory ability of male offspring were detected by open test, target identification test and eight arm labyrinth test; HE staining, transmission electron microscopy, and Rea were used. L-time PCR and Western blotting were used to detect the morphological structure of the brain tissue of the male offspring, the ultrastructure of the hippocampus CA1 region, the epigenetic related enzymes (DNMTs and HDACs), the related molecules of the ERK signaling pathway, the synaptic markers (synaptophysin, postsynaptic density-95, PSD-95), and the glutamate receptors. The expression of protein and protein to explore the molecular mechanism of neurotoxicity induced by BPA exposure during embryonic period. [results] (1) BPA exposure in embryonic stage affects the physiological and biochemical indexes of postnatal male offspring, including increasing the weight of male mice at ninth days, 14 days and 21 days after birth, and reducing the ratio of the visceral body in the brain, hippocampus and testis, and the increase of serum testosterone. At the same time, the serum estrogen level was reduced, and the pathological damage of the hippocampal morphology was induced in male offspring: the pyramidal cells and granulosa cells in the hippocampus were obviously smaller, the number of cells decreased, the intercellular space increased, the neuron crinkled and the cell edge was not clear. (2) the BPA exposure in the embryonic stage affected the male offspring in the opening test after birth. Spontaneous behavior in the test, including reduced activity activity, reduced exploratory behavior and hair handling, impaired short-term and long-term memory in target recognition experiments, increased the number of reference memories in the eight arm maze, especially working memory errors. (3) BPA exposure in the embryonic stage affected the related tables in the hippocampus of postnatal male offspring. The expression of Real-time PCR showed that BPA exposure inhibited the expression level of DNA methyltransferase DNMT1 mRNA and histone deacetylase HDAC1 mRNA, up up the expression level of the histone deacetylase HDAC2 mRNA. (4) BPA exposure at embryo stage affected the expression of ERK pathway related molecules in the hippocampus of postnatal male offspring. Lotting data showed that compared with the control group, the expression level of Akt, phospho-Akt, p44/42 MAPK, phospho-p44/42 MAPK, phospho-CREB, BDNF protein in the hippocampus of the male mice of the BPA exposure group decreased significantly. (5) the BPA exposure in the embryonic stage affected the ultrastructure of the synapse in the hippocampal CA1 area of the male offspring of the postnatal male offspring, including the widening of the synaptic gap, the thinning of the synapse and the synapse. The surface was blurred and synaptic vesicles disappeared; Real-time PCR and Western Blotting experimental data showed that BPA exposure lowered the mRNA and protein expression of the synaptic markers (synaptophysin, PSD-95, and spinophilin), and inhibited the glutamate receptor (G1uR1 and NMDARl) mRNA and protein expression. [Conclusion] the experimental results of this subject revealed the embryo period. BPA exposure induces a decrease in the learning and memory ability of postnatal male offspring, and its molecular mechanism involves epigenetics, ERK pathway and synaptic plasticity, suggesting that BPA affects the complexity of the central nervous system and the mechanism of behavioral development.
【學(xué)位授予單位】：山西農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 陳燕;;神經(jīng)元的突觸可塑性與學(xué)習(xí)和記憶[J];生物化學(xué)與生物物理進(jìn)展;2008年06期

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本文編號(hào)：1988456