本文選題：鋁 + 認(rèn)知功能��； 參考：《山西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:通過體內(nèi)實驗探討線粒體分裂融合在鋁致大鼠認(rèn)知功能障礙的作用。方法:180只清潔級健康雄性SD大鼠,采用隨機(jī)數(shù)字表法按體質(zhì)量隨機(jī)分為5組,即空白組、溶劑對照組、低、中、高劑量組,每組12只。溶劑對照組、低、中、高劑量組分別予劑量為0.41、0.81、1.62 mg/kg體質(zhì)量的麥芽酚鋁溶液,溶劑對照組予等量生理鹽水,空白組不施加任何干預(yù)措施,腹腔注射染毒,隔日注射。大鼠分為3批,第一批染毒一個月,第二批染毒二個月,第三批染毒三個月。染毒結(jié)束后,采用Morris水迷宮實驗測定大鼠空間學(xué)習(xí)記憶能力,采用石墨爐法測定大鼠海馬鋁含量,采用透射電鏡觀察線粒體超微機(jī)構(gòu),采用化學(xué)比色法測定海馬組織線粒體酶活性,采用Western-blot檢測海馬組織CoxⅣ、Drp1、Fis1、Opa1、Mfn1、Mfn2、CaN和S-drp1(s637)蛋白的相對表達(dá)水平。結(jié)果:1.Morris水迷宮實驗檢測結(jié)果:(1)定位航行實驗逃避潛伏期重復(fù)測量方差分析各批大鼠訓(xùn)練時間逃避潛伏期在染鋁劑量與訓(xùn)練時間上交互作用不存在統(tǒng)計學(xué)意義(F1=1.648,P=0.125;F2=0.986,P=0.453;F3=0.763,P=0.664)。三批老鼠通過訓(xùn)練,逃避潛伏期均隨訓(xùn)練時間的延長而縮短(F1=182.462,P0.001;F2=307.703,P0.001;F3=110.248,P0.001)。(2)空間探索試驗?zāi)繕?biāo)象限停留時間空間探索實驗大鼠目標(biāo)象限停留時間染鋁劑量與染鋁時間交互作用有統(tǒng)計學(xué)意義(F=4.853,P0.001),隨著染毒時間和染毒劑量的增加,大鼠目標(biāo)象限停留時間均減少(P0.05)。(3)空間探索實驗穿越平臺次數(shù)隨著染毒劑量和染毒時間的增加,大鼠穿越平臺次數(shù)減少。大鼠穿越平臺次數(shù)在染鋁劑量與染鋁時間的交互作用無統(tǒng)計學(xué)意義(F=0.409,P=0.913)。2.石墨爐法測定大鼠海馬腦鋁含量:對大鼠腦鋁水平進(jìn)行鋁暴露時間與劑量的析因方差分析,結(jié)果顯示各鋁暴露時間組間腦鋁差異有統(tǒng)計學(xué)意義(f=9.009;p0.05),后續(xù)進(jìn)行鋁暴露時間組兩兩比較發(fā)現(xiàn)與鋁暴露1個月組相比,2個月組和3個月組腦鋁水平明顯上升(p0.05)。大鼠腦鋁結(jié)果在鋁暴露時間與劑量上并無交互作用(f=1.097;p0.05)。3.透射電鏡線粒體超微結(jié)構(gòu)結(jié)果:染鋁3個月大鼠高劑量組海馬組織線粒體與空白組、溶劑對照組相比多出現(xiàn)腫脹,內(nèi)棘大部分排列紊亂,有些內(nèi)部出現(xiàn)空泡化,線粒體棘溶解,線粒體變大變圓隨時有破裂的跡象。4.化學(xué)比色法測定海馬組織線粒體酶活性結(jié)果:(1)對三批次各劑量組大鼠海馬na+-k+atp酶活性表達(dá)量析因分析,結(jié)果表明na+-k+atp酶活性表達(dá)量在染鋁劑量與染鋁時間交互作用有統(tǒng)計學(xué)意義(f=2.675,p=0.012)。因此隨著染毒劑量的增加,na+-k+atp酶活性降低(p0.05),僅中劑量和高劑量組染毒三個月與染毒兩個月差別無統(tǒng)計學(xué)意義,各組比較差異均具有統(tǒng)計學(xué)意義。(2)對三批次各劑量組大鼠海馬ca2+-mg2+atp酶活性表達(dá)量析因分析,結(jié)果表明ca2+-mg2+atp酶活性表達(dá)量在染鋁劑量與染鋁時間交互作用有統(tǒng)計學(xué)意義(f=3.665,p0.001)。隨著染毒劑量的增加,ca2+-mg2+atp酶活性降低(p0.05)。5.western-blot檢測海馬組織coxⅣ、drp1、fis1、opa1、mfn1、mfn2、can和s-drp1(s637)蛋白的相對表達(dá)水平結(jié)果:(1)coxⅣ對三批次各劑量組大鼠海馬coxⅣ表達(dá)量析因分析,結(jié)果表明coxⅣ蛋白表達(dá)量在染鋁劑量與染鋁時間交互作用有統(tǒng)計學(xué)意義(f=2.11,p0.05),隨著染毒劑量的增加,蛋白表達(dá)量降低(p0.05)。(2)drp1對三批次各劑量組大鼠海馬drp1表達(dá)量析因分析,結(jié)果表明drp1蛋白表達(dá)量在染鋁劑量與染鋁時間交互作用有統(tǒng)計學(xué)意義(f=4.306,p0.001)。隨著染毒劑量的增加,蛋白表達(dá)量增加(p0.05)。(3)fis1對三批次各劑量組大鼠海馬fis1表達(dá)量析因分析,結(jié)果表明fis1蛋白表達(dá)量在染鋁劑量與染鋁時間交互作用無統(tǒng)計學(xué)意義(f=1.168,p=0.330)。高劑量組fis1蛋白表達(dá)量分別比空白組、溶劑對照組和低劑量組蛋白表達(dá)量增多0.19倍、0.16倍和0.14倍。(4)opa1、mfn1、mfn2對三批次各劑量組大鼠海馬opa1表達(dá)量析因分析,結(jié)果表明opa1、mfn1、mfn2蛋白表達(dá)量在染鋁劑量與染鋁時間交互作用無統(tǒng)計學(xué)意義(f=1.942,p=0.066)、(f=1.012,p=0.435)、(f=0.255,p=0.978)。高劑量組大鼠opa1蛋白表達(dá)量比空白組、溶劑對照組、低劑量組、中劑量分別增加1.12倍、1.07倍、0.39倍、0.22倍;高劑量組大鼠Mfn1蛋白表達(dá)量比空白組、溶劑對照組、低劑量組、中劑量分別增加0.71倍、0.70倍、0.47倍、0.21倍;高劑量組Mfn2蛋白表達(dá)量分別比空白組、溶劑對照組和低劑量組蛋白表達(dá)量增多0.41倍、0.37倍和0.2倍。(5)S-drp1(s637)對三批次各劑量組大鼠海馬s-Drp1(637)表達(dá)量析因分析,結(jié)果表明s-Drp1(s637)蛋白表達(dá)量在染鋁劑量與染鋁時間交互作用有統(tǒng)計學(xué)意義(F=2.477,P=0.019)隨著染毒劑量的增加,蛋白表達(dá)量降低,染毒三個月時高劑量組蛋白表達(dá)量相比溶劑對照組、低劑量、中劑量分別降低了67.5%、51.8%、21.2%。(6)對三批次各劑量組大鼠海馬CaN表達(dá)量析因分析,結(jié)果表明CaN蛋白表達(dá)量在染鋁劑量與染鋁時間交互作用有統(tǒng)計學(xué)意義(F=2.904,P=0.047)。隨著染毒劑量的增加,蛋白表達(dá)量增加,染毒三個月時高劑量組蛋白表達(dá)量相比溶劑對照組、低劑量、中劑量分別增加了0.64倍、0.31倍、0.10倍。結(jié)論:1.亞慢性染鋁可導(dǎo)致大鼠學(xué)習(xí)記憶能力損傷,致大鼠認(rèn)知功能障礙,且存在劑量及時間依賴性。2.鋁暴露可致大鼠海馬線粒體損傷,線粒體功能下降,且存在時間劑量依賴性。3.鋁暴露可打破大鼠海馬線粒體分裂融合平衡,且以打破線粒體分裂為主,具有時間劑量依賴性。綜上所述,鋁可能通過打破線粒體分裂融合平衡進(jìn)而引起線粒體損傷、功能下降,最終會導(dǎo)致大鼠認(rèn)知功能障礙。
[Abstract]:Objective: To investigate the effect of mitochondrial mitotic fusion on cognitive dysfunction induced by aluminum in rats. Methods: 180 healthy male SD rats were randomly divided into 5 groups by random number table method according to body mass, namely, blank group, solvent control group, low, middle and high dose group, with 12 rats in each group. The group of low, middle and high dose groups were given respectively. The dose of 0.41,0.81,1.62 mg/kg body mass of Maltol aluminum solution, the solvent control group was given the same amount of physiological saline, the blank group did not exert any intervention measures, intraperitoneal injection and daily injection. The rats were divided into 3 batches, the first batch was poisoned for one month, the second batches were poisoned for two months, and the third batches were poisoned for three months. After the end of the poisoning, the Morris water maze was used. The spatial learning and memory ability of rats was measured. The content of aluminum in the hippocampus of rats was measured by graphite furnace method. The mitochondrial ultrastructure was observed by transmission electron microscopy. The activity of mitochondrial enzyme in the hippocampus was measured by chemical colorimetry. The relative expression of Cox IV, Drp1, Fis1, Opa1, Mfn1, Mfn2, CaN and S-drp1 (s637) protein in the hippocampus was detected by Western-blot. Results: results: the results of 1.Morris water maze test: (1) the analysis of variance analysis of the escape incubation period of the navigation experiment, there is no statistical significance (F1=1.648, P=0.125; F2= 0.986, P=0.453; F3=0.763, P=0.664) in the training time of the training time of each batch of rats in the escape incubation period of the training time of each batch of rats (P=0.125, F3=0.763, P=0.664). The incubation period shortened with the prolongation of the training time (F1=182.462, P0.001; F2=307.703, P0.001; F3=110.248, P0.001). (2) space exploration test target quadrant time space exploration of experimental rats' target quadrant time, the interaction between aluminum dye and aluminum dyeing time was statistically significant (F=4.853, P0.001), with the time of exposure and exposure. The residence time of the target quadrant of rats decreased (P0.05). (3) the number of crossing platform in rats decreased with the increase of exposure dose and time. The interaction between the number of rat crossing platform and the interaction between the dose of aluminum dye and the time of aluminum dyeing was not statistically significant (F=0.409, P=0.913).2. graphite furnace method The content of aluminum in the hippocampus of rats was determined by the analysis of variance analysis of aluminum exposure time and dose of aluminum exposure in rats. The results showed that there was a significant difference between the aluminum exposure time group and the aluminum exposure group (f=9.009; P0.05). Compared with the aluminum exposure group 22, the aluminum exposure was compared with the 1 months of aluminum exposure group, and the 2 months group and 3 month group of brain al water were clear. There was a significant increase (P0.05). There was no interaction between the aluminum exposure time and the dose of aluminum exposure (f=1.097; P0.05). The ultrastructural results of mitochondrial ultrastructure of.3. transmission electron microscopy: the hippocampus mitochondria and the blank group in the high dose group of 3 months of aluminum infected rats, the swelling, the disorder of the most of the internal spines, and the vacuolation in some of the internal spines were found in the high dose group. The mitochondrial spines dissolved, the mitochondria became large and round and there were signs of rupture at any time..4. chemical colorimetry was used to determine the results of mitochondrial enzyme activity in the hippocampus: (1) analysis of the expression of na+-k+atp enzyme activity in the hippocampus of three batches of rats. The results showed that the interaction of na+-k+atp enzyme activity expression at the dose of aluminum and the time of aluminum dyeing was statistically significant. Meaning (f=2.675, p=0.012). Therefore, with the increase of dose, the activity of na+-k+atp enzyme decreased (P0.05). There was no statistically significant difference between the middle dose and high dose group for three months and two months. (2) the analysis of ca2+-mg2+atp enzyme activity expression in the hippocampus of three batches of rats The results showed that the interaction of the activity of ca2+-mg2+atp enzyme activity was statistically significant (f=3.665, p0.001). With the increase of dose, the activity of ca2+-mg2+atp enzyme decreased (P0.05).5.western-blot to detect the relative expression level of Cox IV, drp1, FIS1, OPA1, mfn1, Mfn2, Mfn2, drp1, and.5.western-blot in the hippocampus. (1) analysis of Cox IV expression in hippocampus of rats in each dose group of Cox IV to three batches. The results showed that the expression of Cox IV protein expression was statistically significant (f=2.11, P0.05) at the dose of aluminum dye and aluminum dyeing time (f=2.11, P0.05). The expression of protein decreased with the increase of dose (P0.05). (2) the expression of drp1 in the hippocampus of each dose group of three batches of drp1 was analyzed by drp1 The results showed that the interaction of drp1 protein expression and aluminum dyeing time was statistically significant (f=4.306, p0.001). As the dose increased, the protein expression increased (P0.05). (3) the analysis of FIS1 expression in hippocampal FIS1 of FIS1 to three batches of rats showed that the expression of FIS1 protein was in the dose of aluminum and aluminum. The time interaction was not statistically significant (f=1.168, p=0.330). The expression of FIS1 protein in the high dose group was 0.19 times more than that in the blank group, the expression of the protein in the solvent control group and the low dose group increased by 0.19 times, 0.16 times and 0.14 times. (4) OPA1, mfn1, Mfn2 were analyzed for the expression of OPA1 in the hippocampus of three batches of rats. The results showed that the expression of OPA1, mfn1, Mfn2 protein was expressed. There was no statistical significance (f=1.942, p=0.066), (f=1.012, p=0.435), (f=1.012, p=0.435), (f=0.255, p=0.978). The expression of OPA1 protein in the high dose group was 1.12 times more, 1.07 times, 0.39 times, 0.22 times than that in the blank group, the low dose group and the low dose group respectively. The expression of Mfn1 protein in the high dose group was more than that of the blank group. In the solvent control group, the low dose group increased 0.71 times, 0.70 times, 0.47 times, 0.21 times respectively. The expression of Mfn2 protein in the high dose group was 0.41 times, 0.37 times and 0.2 times more than that in the blank group, 0.37 times and 0.2 times in the solvent control group and the low dose group. (5) the expression of the s-Drp1 (637) in the hippocampus of the three batches of rats was analyzed, the result of the analysis of the results of the expression of the s-Drp1 (637) in the hippocampus of three batches of rats. The results showed that the expression of s-Drp1 (s637) protein expression was statistically significant (F=2.477, P=0.019), with the increase of the dose, the protein expression decreased, and the high dose of protein expression was compared with the solvent control group at three months. The low dose and middle dose decreased by 67.5%, 51.8%, and 21.2%. (6) to three batches, respectively. The analysis of CaN expression in the hippocampus of dose group showed that the expression of CaN protein expression was statistically significant (F=2.904, P=0.047) in the dose of aluminum dyed and aluminum dye (F=2.904, P=0.047). With the increase of dose, the protein expression increased, and the high dose of protein expression was compared with the solvent control group at three months, and the low dose and middle dose increased respectively. The addition of 0.64 times, 0.31 times, and 0.10 times. Conclusion: 1. subchronic aluminum dyed aluminum can lead to the impairment of learning and memory ability in rats, and induce cognitive dysfunction in rats. There is a dose and time dependent.2. aluminum exposure that can cause damage to the mitochondria of the hippocampus in rats and the decrease of mitochondrial function, and the presence of time dependent.3. aluminum exposure can break the mitochondria of the hippocampus of rats. In summary, aluminum may cause mitochondrial damage by breaking mitochondrial fission and fusion, resulting in mitochondrial damage and decreased function, which eventually leads to cognitive dysfunction in rats.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 Wafa Kharroubi;Samia Hai Ahmed;Thomas Nury;Pierre Andreoletti;Rachid Sakly;Mohamed Hammami;Gerard Lizard;;Mitochondrial dysfunction,oxidative stress and apoptotic induction in microglial BV-2 cells treated with sodium arsenate[J];Journal of Environmental Sciences;2017年01期

2 亢盼;李朝陽;牛僑;;亞慢性鋁暴露對大鼠學(xué)習(xí)記憶能力及組蛋白H3K4甲基轉(zhuǎn)移酶影響研究[J];中國職業(yè)醫(yī)學(xué);2015年05期

3 趙若聰;劉瓊;何曉陽;;線粒體動力學(xué)失衡和環(huán)境神經(jīng)毒物對阿爾茨海默病病理進(jìn)程影響的研究進(jìn)展[J];中國藥理學(xué)與毒理學(xué)雜志;2013年01期

4 李文偉;朱敏;呂傳真;;線粒體動力學(xué)改變在神經(jīng)變性疾病中的地位[J];生理科學(xué)進(jìn)展;2011年05期

相關(guān)博士學(xué)位論文 前1條

1 羅磊;“益腎調(diào)督”法對AD模型大鼠神經(jīng)元軸突線粒體損傷的影響及針灸作用機(jī)制研究[D];湖北中醫(yī)藥大學(xué);2014年

，

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