發(fā)布時間：2018-05-07 22:06

  本文選題：錳中毒 + CREB��； 參考：《廣西醫(yī)科大學》2013年碩士論文

【摘要】：目的：觀察豐富環(huán)境和貧瘠環(huán)境干預對錳中毒小鼠學習記憶能力和神經發(fā)生的影響,并初步探討其作用機制,為錳中毒的防治提供理論基礎和實驗依據(jù)。 方法：選用清潔級成年昆明小鼠雌性40只,體重20±2g,隨機分成空白對照組(CG)、正常環(huán)境染錳組(SEG)、豐富環(huán)境干預染錳組(EEG)以及貧瘠環(huán)境干預染錳組(IEG),共4組,每組10只。采用腹腔注射氯化錳(MnCl220mg/kg)的方式制造小鼠錳中毒模型,進行染毒3周,而對照組腹腔注射同等劑量的生理鹽水,同時各組小鼠在相應的環(huán)境中進行刺激。用Morris水迷宮的方法檢測各組小鼠的學習記憶能力,用免疫熒光化學法檢測小鼠SVZ和SGZ內DCX的表達,用免疫組織化學的方法檢測海馬內CREB蛋白的表達。用Image-Pro Plus6.0圖像分析系統(tǒng)計算SVZ和SGZ內DCX的陽性表達的平均光密度值及海馬區(qū)CREB陽性表達的平均光密度值,用SPSS軟件對所有數(shù)據(jù)做統(tǒng)計分析,并作線性相關分析。 結果：1.水迷宮定位航行試驗中正常環(huán)境染錳組小鼠的逃避潛伏期比空白對照組小鼠的明顯延長(p0.05),而豐富環(huán)境干預染錳組小鼠的逃避潛伏期明顯比正常環(huán)境干預染錳組小鼠的縮短(p0.05),貧瘠環(huán)境干預染錳組與正常環(huán)境干預染錳組之間無統(tǒng)計學差異；空間探索實驗中,正常環(huán)境染錳組小鼠穿越平臺的次數(shù)比空白對照組少,豐富環(huán)境干預染錳組穿越平臺的次數(shù)則比正常環(huán)境染錳組多,貧瘠環(huán)境干預染錳組比正常環(huán)境染錳組少,差異有統(tǒng)計學意義(p0.05)。2.DCX免疫組織化學檢測結果顯示,正常環(huán)境干預染錳組和貧瘠環(huán)境干預染錳組SVZ和SGZ區(qū)DCX陽性熒光強度明顯比對照組弱；豐富環(huán)境干預染錳組SVZ和SGZ區(qū)DCX陽性熒光強度明顯比正常環(huán)境染錳組強,差異有統(tǒng)計學意義,與對照組比較無統(tǒng)計學差異。3.CREB免疫組織化學檢測結果顯示,正常環(huán)境染錳組和貧瘠環(huán)境干預染錳組海馬CA1區(qū)CREB蛋白表達量明顯低于對照組(p0.01)；豐富環(huán)境干預染錳組海馬CA1區(qū)內CREB蛋白表達量比正常環(huán)境干預染錳組的要明顯增多(p0.01),與對照組無統(tǒng)計學差異。4.線性相關分析表明,CREB的表達與小鼠記憶能力及SGZ內DCX的表達呈正相關。 結論：1.豐富環(huán)境干預可改善錳中毒小鼠的學習記憶能力。2.豐富環(huán)境干預可促進錳中毒小鼠腦內的神經發(fā)生。3.豐富環(huán)境干預可增加海馬內CREB的表達。4.貧瘠環(huán)境干預降低錳中毒小鼠的學習記憶能力,抑制了腦內的神經發(fā)生,降低了錳中毒小鼠的海馬內的CREB的表達。5.豐富環(huán)境干預改善錳中毒小鼠學習記憶能力促進腦內神經發(fā)生的機制,可能與豐富環(huán)境干預增加海馬內CREB的表達有關。
[Abstract]:Objective: to observe the effects of rich and barren environment intervention on learning and memory ability and neurogenesis in mice with manganese poisoning, and to explore its mechanism, and to provide theoretical basis and experimental basis for prevention and treatment of manganese poisoning. Methods: forty adult Kunming mice of clean grade, weighing 20 鹵2 g, were randomly divided into 4 groups: control group (10 rats), normal environment exposed group (EEGG), enriched environment group (EEGG) and barren environment group (10 rats in each group). The mice model of manganese poisoning was made by intraperitoneal injection of MnCl _ 220mg / kg. The mice in the control group were injected with the same dose of normal saline for 3 weeks, and the mice in each group were stimulated in the corresponding environment. The ability of learning and memory was detected by Morris water maze method, the expression of DCX in SVZ and SGZ was detected by immunofluorescence method, and the expression of CREB protein in hippocampus was detected by immunohistochemical method. The average optical density of DCX positive expression in SVZ and SGZ and the average optical density of CREB positive expression in hippocampus were calculated by Image-Pro Plus6.0 image analysis system. All the data were statistically analyzed by SPSS software and linear correlation analysis was made. The result is 1: 1. In the water maze navigation test, the escape latency of the mice exposed to manganese in the normal environment was significantly longer than that of the control group, while the escape latency of the mice in the enriched environment group was significantly smaller than that in the control group. There was no significant difference between the barren environmental intervention group and the normal environmental intervention group. In the space exploration experiment, the mice in the normal environment exposed to manganese had less times of crossing the platform than those in the blank control group, and the rich environment intervention group had more times of crossing the platform than that of the normal environment group. The poor environment intervention group was less than that in the normal environment group, and the difference was statistically significant (P 0.05). 2. The results of DCX immunohistochemical analysis showed that there was no significant difference between the two groups. The positive fluorescence intensity of DCX in the SVZ and SGZ regions in the normal and barren environment groups was significantly weaker than that in the control group, and the positive fluorescence intensity of DCX in the SVZ and SGZ regions in the rich environment group was significantly stronger than that in the normal environment group. The difference was statistically significant. There was no significant difference between the two groups. 3. The results of CREB immunohistochemistry showed that, The expression of CREB protein in the CA1 region of hippocampus in the manganese exposed group and the barren environment group was significantly lower than that in the control group, while the CREB protein expression in the CA1 region in the rich environment group was significantly higher than that in the control group. There was no significant difference between the control group and the control group. Linear correlation analysis showed that the expression of CREB was positively correlated with the memory ability of mice and the expression of DCX in SGZ. Conclusion 1. Rich environment intervention can improve the learning and memory ability of mice exposed to manganese. 2. 2. Rich environment intervention can promote neurogenesis. 3. 3 in the brain of mice with manganese poisoning. Rich environment intervention increased the expression of CREB. 4. 4. The barren environment intervention decreased the learning and memory ability, inhibited the neurogenesis in the brain and decreased the expression of CREB in hippocampus of the mice exposed to manganese. The mechanism of rich environment intervention to improve the learning and memory ability of mice induced by manganese poisoning may be related to the increase of CREB expression in hippocampus.
【學位授予單位】：廣西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R114

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本文編號：1858648