本文選題：鉛暴露 + 神經(jīng)退行性病變　； 參考：《南昌大學(xué)》2013年碩士論文

【摘要】：目的：研究表明鉛不僅具有神經(jīng)發(fā)育毒性,而且可能是神經(jīng)退行性疾病的一個(gè)重要的危險(xiǎn)因素,本文旨在探討長(zhǎng)期鉛暴露致大鼠神經(jīng)退行性改變及其相關(guān)的基因蛋白表達(dá)研究。方法：將成年雌性Sprague-Dawley (SD)大鼠隨機(jī)分為3個(gè)組：空白組、低鉛暴露組(LLG)和高鉛暴露組(HLG),分別給予0,800,1500mg/L乙酸鉛水溶液,染毒10天后與自由飲用去離子水的雄性大鼠合籠。在觀察到陰栓后21天,仔鼠降生,選擇雄性仔鼠分別飼養(yǎng)至斷乳(Postnatal week 3, PNW3)、中年(Postnatal week 41, PNW41)、老年(Postnatal week 70, PNW70)。雄性仔鼠在三個(gè)組分別給予0,300,900mg/L乙酸鉛水溶液。染毒末,通過(guò)活體磁共振成像(Magnetic resonance imaging, MRI)技術(shù)測(cè)量老年大鼠大腦及海馬容積；用離體電感耦合等離子體原子發(fā)射光譜(Inductively coupled plasma atomic emission spectrum-etry, ICP-AES)方法測(cè)定大鼠三個(gè)年齡段血鉛和皮層、海馬鉛含量；硫堇染色觀察三個(gè)年齡段皮層、海馬神經(jīng)元密度情況；采用透射電子顯微鏡觀察老年大鼠海馬神經(jīng)元超微結(jié)構(gòu)損傷情況；采用酶聯(lián)免疫吸附試驗(yàn)(Enzyme-linked immunosorbent assay, ELIS A)方法測(cè)量大腦DNA氧化損傷指標(biāo)8。羥基脫氧鳥苷(8-hydroxy2'deoxyguanosine,8-OHdG)含量；采用實(shí)時(shí)熒光定量PCR (Quantitative Real Time PCR, qRT-PCR)和Western blot方法進(jìn)行淀粉樣前體蛋白(Amyloid precursor protein, APP)、tau、葡萄糖反應(yīng)蛋白78(Glucose-regulated protein 78, GRP78)、葡萄糖反應(yīng)蛋白94 (Glucose-regulated protein 94, GRP94)的表達(dá)量分析。結(jié)果：(1)各年齡段血鉛及腦組織鉛含量均隨著鉛暴露劑量的增加而增加,呈現(xiàn)劑量-反應(yīng)關(guān)系,且均有統(tǒng)計(jì)學(xué)差異；各組在不同年齡段的血鉛無(wú)顯著性差異,但皮層、海馬中鉛在中年與老年期均比斷乳期高,均有顯著性差異。(2)鉛暴露大鼠形態(tài)學(xué)改變：活體MRI結(jié)果顯示,鉛暴露老年大鼠大腦容積下降(pD.D5),海馬容積呈下降趨勢(shì),但無(wú)統(tǒng)計(jì)學(xué)意義；離體硫堇染色結(jié)果可知,在斷乳期,鉛暴露組海馬CA1區(qū)神經(jīng)元密度下降,在老年期,鉛暴露組大鼠皮層頂區(qū)和海馬CA1區(qū)神經(jīng)元密度明顯下降,且與空白組比較有統(tǒng)計(jì)學(xué)差異,海馬DG區(qū)呈下降趨勢(shì)；電鏡結(jié)果提示,隨著染鉛劑量增加,胞質(zhì)、胞核、內(nèi)質(zhì)網(wǎng)結(jié)構(gòu)和突觸結(jié)構(gòu)均遭到不同程度的破壞,出現(xiàn)早期神經(jīng)元細(xì)胞凋亡現(xiàn)象。(3)在斷乳和老年期,鉛暴露使大腦皮層、海馬APP表達(dá)增加,在中年有上升趨勢(shì),但差異無(wú)統(tǒng)計(jì)學(xué)意義；在斷乳和老年期,鉛暴露使海馬tau及磷酸化tau (Ser396)蛋白表達(dá)增加,但在大鼠皮層tau表達(dá)無(wú)差異。(4)鉛暴露增加了斷乳和老年大鼠大腦8-OHdG的含量,且呈劑量-反應(yīng)關(guān)系；但在中年變化不明顯。(5)在老年期,鉛暴露使大腦皮層、海馬GRP78表達(dá)上升,在斷乳和中年期變化均不明顯；在斷乳和老年期,海馬各組GRP94表達(dá)呈上升趨勢(shì),但在皮層變化不明顯,中年變化均不明顯。結(jié)論：(1)大鼠形態(tài)學(xué)結(jié)果和阿爾茨海默病特征性基因上調(diào)提示鉛暴露大鼠出現(xiàn)了神經(jīng)退行性改變。(2)鉛暴露導(dǎo)致的神經(jīng)退行性改變可能與增加了內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)有關(guān),也可能與增加了APP/tau的表達(dá)有關(guān)。(3)長(zhǎng)期鉛暴露大鼠各指標(biāo)變化具有年齡階段性。
[Abstract]:Objective: the study shows that lead not only has neurodevelopmental toxicity, but also may be an important risk factor for neurodegenerative disease. This paper aims to investigate the neurodegenerative changes and related gene protein expression in rats induced by long-term lead exposure. Methods: adult female Sprague-Dawley (SD) rats were randomly divided into 3 groups: empty In the white group, the low lead exposure group (LLG) and the high lead exposure group (HLG) were given 08001500mg/L lead acetic acid solution respectively, and the male rats who drank the deionized water for 10 days were caged. The offspring were born 21 days after the observation of the suppository, and the male offspring were fed to the weaning (Postnatal week 3, PNW3), middle age (Postnatal week 41, PNW41), old age, and old. (Postnatal week 70, PNW70). Male offspring rats were given 0300900mg/L acetate aqueous solution in three groups. The volume of brain and hippocampus in old rats was measured by living magnetic resonance imaging (Magnetic resonance imaging, MRI), and the isolated atomic emission spectrum (Inductively coupled plasma atom) was used as an isolated inductor coupling. IC emission spectrum-etry, ICP-AES) method was used to determine the blood lead and cortex and hippocampus lead content in three age rats. The density of hippocampal neurons in three age groups was observed by thio staining. The ultrastructural damage of hippocampal neurons in the aged rats was observed by transmission electron microscope, and the enzyme linked immunosorbent assay (Enzyme-li) was used. Nked immunosorbent assay, ELIS A) method was used to measure the content of 8. hydroxyl deoxy guanosine (8-hydroxy2'deoxyguanosine, 8-OHdG) of DNA oxidative damage in the brain. The expression of protein 78 (Glucose-regulated protein 78, GRP78) and glucose reactive protein 94 (Glucose-regulated protein 94, GRP94) was analyzed. Results: (1) the lead content of blood and brain tissue in all age groups increased with the increase of lead exposure dose, showing a dose response relationship, and there were statistical differences; each group was in different ages. There was no significant difference in blood lead in the segment, but in the cortex and the hippocampus, lead in the hippocampus was higher in the middle age and the aged than in the weaning period. (2) the morphological changes in the exposed rats showed that the volume of the brain in the aged rats decreased (pD.D5), the volume of the hippocampus decreased, but the volume of the hippocampus decreased, but the result of the isolated thio staining could be found. In the weaning period, the density of neurons in the hippocampal CA1 area decreased in the lead exposed group. In the elderly, the density of neurons in the cortex and hippocampus CA1 area of the lead exposed rats decreased significantly, and there was a statistical difference between the group and the blank group. The hippocampal DG area decreased. The electron microscope results suggested that the cytoplasm, the nucleus, the endoplasmic reticulum structure and the process of the endoplasmic reticulum were increased with the increase of the dose of lead. The contact structure was damaged in different degrees and appeared early neuronal apoptosis. (3) in weaning and old age, lead exposure increased the expression of APP in the cerebral cortex and hippocampus, and increased in middle age, but the difference was not statistically significant; in weaning and old age, the expression of tau and phosphorylated tau (Ser396) protein in the hippocampus increased, but in the weaning and old age, the expression of the protein in the hippocampus and phosphorylated tau (Ser396) was increased, but in the weaning and the old age, the expression of the protein was increased. There was no difference in the expression of tau in the cortex of rats. (4) lead exposure increased the content of 8-OHdG in the brain of weaning and old rats, and showed a dose response relationship, but the change in middle age was not obvious. (5) the expression of lead in the cerebral cortex and hippocampal GRP78 increased in the old age, and the changes in the weaning and middle age were not obvious; in the weaning and the old age, the hippocampus was GRP94 The expression showed an upward trend, but the changes in the cortex were not obvious, and the changes in middle age were not obvious. Conclusion: (1) the morphological results of the rats and the up-regulated genes of Alzheimer's disease suggest that the lead exposed rats have neurodegenerative changes. (2) the neurodegenerative changes caused by lead exposure may be related to the increase of endoplasmic reticulum stress response, and may also be associated with the increase of the endoplasmic reticulum stress response. It was related to the increase of APP/tau expression. (3) the changes of each indicator in the long-term lead exposed rats were of age.

【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R114

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本文編號(hào)：1791812