小鼠慢性酒精中毒脑神经细胞APP和Aβ蛋白的表达
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摘要
前言
我国是一个人口大国,而且是一个老龄化进程最快、老龄人口众多的国家。我国又是一个具有悠久历史、多种多样酒文化并存的国家。由于特殊的社会和历史原因,酒精中毒的情况经常发生。
慢性酒精中毒(chronic alcoholism)是以对乙醇的成瘾为特征,由于经常反复饮酒对乙醇产生耐受,当其戒酒时出现脱瘾(withdrawal)的神经症候。慢性酒精中毒引起神经系统及其他器官的严重损害,其中神经系统的损害最为严重。在法医学实践中,有时会遇到酒精中毒的案例,常与交通事故、重要器官组织功能障碍等有关。
阿尔茨海默病(Alzheimer's disease,AD)是一种渐进性的神经变性性疾病,表现为全面的认知功能障碍。主要病理学改变是由来源于淀粉样前体蛋白(amyloid precursor protein,APP)的β-淀粉样肽(beta-amyloid,Aβ)构成的淀粉样斑块和过度或异常磷酸化的Tau蛋白构成的神经纤维缠结。
阿尔茨海默病的发病受很多因素影响,包括环境因素、不良饮食习惯、药物种类、中毒等。慢性酒精中毒是否会导致脑内APP和Aβ蛋白的增多从而引发阿尔茨海默病的发生,目前尚缺乏系统的理论和实验研究。
神经细胞特异性烯醇化酶(Neuron-specific enolase,NSE)是神经细胞的标志性蛋白,特异性的存在于神经元和神经内分泌细胞中,占脑内全部可溶性蛋白的1.5%,神经元损伤或坏死后NSE进入血液。
本研究在建立小鼠慢性酒精中毒的动物模型基础上,应用组织病理学和免疫组织化学等技术,研究小鼠慢性酒精中毒后,脑组织神经细胞中APP和Aβ的表达,并应用SPSS统计软件对观察到的实验结果进行统计学分析,探讨慢性酒精中毒脑神经细胞中APP和Aβ的含量变化以及与阿尔茨海默病发病之间的因果关系。
材料和方法
一、动物模型的建立及分组
雄性健康9周龄昆明小鼠42只,随机分为30天组,60天组,每组又分10%酒精组、20%酒精组和对照组,其中对照组各5只,10%酒精组各8只,20%酒精组各8只。
实验组将无水乙醇(色谱纯)用饮用矿泉水配成浓度为10%、20%酒精溶液作为实验小鼠饮用水,对照组采用饮用矿泉水代替。实验结束后,直接断头处死动物,迅速提取小鼠脑组织,4%多聚甲醛/PBS pH7.4固定液固定,石蜡包埋、制片。
二、实验方法
(1)应用HE染色法观察小鼠慢性酒精中毒后脑神经细胞的病理组织学形态变化;
(2)应用免疫组织化学染色法观察小鼠慢性酒精中毒后脑神经细胞内神经元特异性烯醇化酶表达的变化;
(3)应用免疫组织化学染色法观察小鼠慢性酒精中毒后脑神经细胞内APP和Aβ表达情况;
(4)采用Motic Images Advanced 3.2图像分析系统计算免疫组织化学染色阳性细胞数和细胞内阳性反应产物的光密度;
(5)应用SPSS统计学软件对实验数据进行统计分析。
实验结果
(1)各实验组小鼠出现明显的慢性中毒表现,且体重明显下降;
(2) HE染色见慢性酒精中毒的小鼠脑组织水肿,神经细胞内尼氏体消失或边聚,神经细胞周围间隙增宽;
(3)神经元特异性烯醇化酶免疫组织化学染色见该酶蛋白含量增加;
(4)对照组可见少量APP免疫组织化学染色阳性的神经细胞,染色产物分布于细胞胞浆内,呈棕黄色。实验组APP染色阳性细胞明显增多,表达增强,且不同浓度不同时间段阳性表达强度不同。实验组脑神经细胞胞浆内出现明显的Aβ阳性染色产物,且高浓度长时间给药组阳性细胞数明显增多,表达增强。使用Motic Image Advanced 3.2图像分析系统计算各组平均积分光密度,并采用SPSS软件进行统计分析,发现实验组和对照组间有明显差异。
结论
1.慢性酒精中毒可引起实验小鼠脑神经细胞呈退行性改变;
2.慢性酒精中毒可引起实验小鼠脑神经细胞神经元特异性烯醇化酶含量增多和分布异常;
3.慢性酒精中毒可引起实验小鼠脑神经细胞胞浆内产生Aβ蛋白,且APP蛋白的表达明显增强。
4.慢性酒精中毒可能是诱发阿尔茨海默病发病的有关因素之一。
Introduction
China is a country with a large population and the aging process is very fast. China is a long history,and diverse cultures co-exist wine in the country.Because of the particular social and historical reasons,alcoholism frequently is the case.
Chronic alcoholism is characterized by addiction,as is often repeated in drinking on the ethanol tolerance,when it occurs when alcohol withdrawal(withdrawal) symptoms of the nerve.Nervous system caused by chronic alcoholism and serious damage to other organs,including nervous system damage to the most serious.Practice in forensic science,and sometimes encounter cases of alcoholism are often associated with traffic accidents,major organ dysfunction and other related organizations.
Alzheimer's disease(AD) is a progressive neurodegenerative disease,manifested as a comprehensive cognitive impairment.The main histopathological changes is derived from amyloid precursor protein(APP) ofβ-amyloid peptide(Aβ) consisting of amyloid plaque and excessive or abnormal phosphorylation of Tau protein composition neurofibrillary tangles.
The incidence of Alzheimer's disease are subject to many factors,including environmental factors,poor dietary habits,drug types,such as poisoning.Chronic alcoholism lead to brain APP and Aβprotein in order to trigger an increase in the incidence of Alzheimer's disease,is still a lack of systematic theoretical and experimental studies.
Neuron-specific enolase(NSE) is a sign of nerve cell protein,the existence of specificity in neurons and neuroendocrine cells in the brain of all soluble protein accounted for 1.5%of neuronal damage or NSE into the blood after necrosis.
In this study,mice in the establishment of animal models of chronic alcoholism, based on the application of histopathological and immunohistochemical techniques, research in mice after chronic alcoholism and brain nerve cells in the expression of APP and Aβ,and the application of SPSS statistical software of the observed experimental results were statistically analyzed to explore neurons in chronic alcoholism APP and Aβ,as well as the content changes in the incidence of Alzheimer's disease and the causal link between them.
Materials and Methods
1.Establishment of animal model and packet
42 9-week-old Kunming mice were randomly divided into 30-day group,the 60-day groups was divided into 10%alcohol group,20%alcohol group and the control group,of which five of the control group,10%ethanol group 8,and 20%alcohol group 8.
The experimental group will be anhydrous ethanol(HPLC pure) in drinking water to match the concentration of 10%,20%alcohol solution as the drinking water of mice in the control group used in place of drinking water.After the end of the experiment, animals were killed directly decapitation,rapid extraction of mouse brain tissue,4% paraformaldehyde/PBS pH7.4 fixed,paraffin-embedded,producer.
2.Methods
(1) HE staining to observe the application of the morphological characteristics of organizations.
(2) the use of Nissl staining of nerve cells circumstances Nissl bodies.
(3) Application of neuron-specific enolase staining patterns characteristic of neurons.
(4) immunohistochemical staining APP and Aβin the brain tissues.
(5) the use of Motic Images Advanced 3.2 image analysis system to calculate immunohistochemical staining positive cells and expression of the area.
(6) the application of SPSS statistical software for statistical analysis of experimental data
Result
(1) The experimental group of mice significantly the performance of chronic poisoning,and decreased body weight.
(2) HE staining of chronic alcoholism see the mice brain edema,nerve cells in the disappearance of Nissl bodies or margination,nerve cells around the gap widened.
(3) neuron-specific enolase immunohistochemistry see increase in enzyme protein content.
(4) the control group shows that a small number of APP positive immunohistochemical staining of nerve cells,a product of the distribution of staining in the cell cytoplasm,showing brown yellow.Experimental group,APP staining positive cells markedly increased expression of different concentrations and different time periods of different intensity of positive expression.Experimental group,nerve cells showed a visible product of Aβ-positive staining,and high concentrations of prolonged treatment group the number of positive cells markedly increased expression.Using Motic Image Advanced 3.2 image analysis system to calculate the average integrated optical density,and the use of SPSS software for statistical analysis and found that the experimental group and the control group there was significant difference.
Conclusion
(1) Chronic alcoholism can cause brain cells of mice degeneration.
(2) Chronic alcoholism can cause brain cells of mice neuron-specific enolase increase in content and distribution of abnormalities.
(3) Chronic alcoholism can cause brain cells of mice produce Aβprotein,and APP protein expression was significantly enhanced.
(4) Chronic alcoholism may be induced by Alzheimer's disease,one of the relevant factors.
我国是一个人口大国,而且是一个老龄化进程最快、老龄人口众多的国家。我国又是一个具有悠久历史、多种多样酒文化并存的国家。由于特殊的社会和历史原因,酒精中毒的情况经常发生。
慢性酒精中毒(chronic alcoholism)是以对乙醇的成瘾为特征,由于经常反复饮酒对乙醇产生耐受,当其戒酒时出现脱瘾(withdrawal)的神经症候。慢性酒精中毒引起神经系统及其他器官的严重损害,其中神经系统的损害最为严重。在法医学实践中,有时会遇到酒精中毒的案例,常与交通事故、重要器官组织功能障碍等有关。
阿尔茨海默病(Alzheimer's disease,AD)是一种渐进性的神经变性性疾病,表现为全面的认知功能障碍。主要病理学改变是由来源于淀粉样前体蛋白(amyloid precursor protein,APP)的β-淀粉样肽(beta-amyloid,Aβ)构成的淀粉样斑块和过度或异常磷酸化的Tau蛋白构成的神经纤维缠结。
阿尔茨海默病的发病受很多因素影响,包括环境因素、不良饮食习惯、药物种类、中毒等。慢性酒精中毒是否会导致脑内APP和Aβ蛋白的增多从而引发阿尔茨海默病的发生,目前尚缺乏系统的理论和实验研究。
神经细胞特异性烯醇化酶(Neuron-specific enolase,NSE)是神经细胞的标志性蛋白,特异性的存在于神经元和神经内分泌细胞中,占脑内全部可溶性蛋白的1.5%,神经元损伤或坏死后NSE进入血液。
本研究在建立小鼠慢性酒精中毒的动物模型基础上,应用组织病理学和免疫组织化学等技术,研究小鼠慢性酒精中毒后,脑组织神经细胞中APP和Aβ的表达,并应用SPSS统计软件对观察到的实验结果进行统计学分析,探讨慢性酒精中毒脑神经细胞中APP和Aβ的含量变化以及与阿尔茨海默病发病之间的因果关系。
材料和方法
一、动物模型的建立及分组
雄性健康9周龄昆明小鼠42只,随机分为30天组,60天组,每组又分10%酒精组、20%酒精组和对照组,其中对照组各5只,10%酒精组各8只,20%酒精组各8只。
实验组将无水乙醇(色谱纯)用饮用矿泉水配成浓度为10%、20%酒精溶液作为实验小鼠饮用水,对照组采用饮用矿泉水代替。实验结束后,直接断头处死动物,迅速提取小鼠脑组织,4%多聚甲醛/PBS pH7.4固定液固定,石蜡包埋、制片。
二、实验方法
(1)应用HE染色法观察小鼠慢性酒精中毒后脑神经细胞的病理组织学形态变化;
(2)应用免疫组织化学染色法观察小鼠慢性酒精中毒后脑神经细胞内神经元特异性烯醇化酶表达的变化;
(3)应用免疫组织化学染色法观察小鼠慢性酒精中毒后脑神经细胞内APP和Aβ表达情况;
(4)采用Motic Images Advanced 3.2图像分析系统计算免疫组织化学染色阳性细胞数和细胞内阳性反应产物的光密度;
(5)应用SPSS统计学软件对实验数据进行统计分析。
实验结果
(1)各实验组小鼠出现明显的慢性中毒表现,且体重明显下降;
(2) HE染色见慢性酒精中毒的小鼠脑组织水肿,神经细胞内尼氏体消失或边聚,神经细胞周围间隙增宽;
(3)神经元特异性烯醇化酶免疫组织化学染色见该酶蛋白含量增加;
(4)对照组可见少量APP免疫组织化学染色阳性的神经细胞,染色产物分布于细胞胞浆内,呈棕黄色。实验组APP染色阳性细胞明显增多,表达增强,且不同浓度不同时间段阳性表达强度不同。实验组脑神经细胞胞浆内出现明显的Aβ阳性染色产物,且高浓度长时间给药组阳性细胞数明显增多,表达增强。使用Motic Image Advanced 3.2图像分析系统计算各组平均积分光密度,并采用SPSS软件进行统计分析,发现实验组和对照组间有明显差异。
结论
1.慢性酒精中毒可引起实验小鼠脑神经细胞呈退行性改变;
2.慢性酒精中毒可引起实验小鼠脑神经细胞神经元特异性烯醇化酶含量增多和分布异常;
3.慢性酒精中毒可引起实验小鼠脑神经细胞胞浆内产生Aβ蛋白,且APP蛋白的表达明显增强。
4.慢性酒精中毒可能是诱发阿尔茨海默病发病的有关因素之一。
Introduction
China is a country with a large population and the aging process is very fast. China is a long history,and diverse cultures co-exist wine in the country.Because of the particular social and historical reasons,alcoholism frequently is the case.
Chronic alcoholism is characterized by addiction,as is often repeated in drinking on the ethanol tolerance,when it occurs when alcohol withdrawal(withdrawal) symptoms of the nerve.Nervous system caused by chronic alcoholism and serious damage to other organs,including nervous system damage to the most serious.Practice in forensic science,and sometimes encounter cases of alcoholism are often associated with traffic accidents,major organ dysfunction and other related organizations.
Alzheimer's disease(AD) is a progressive neurodegenerative disease,manifested as a comprehensive cognitive impairment.The main histopathological changes is derived from amyloid precursor protein(APP) ofβ-amyloid peptide(Aβ) consisting of amyloid plaque and excessive or abnormal phosphorylation of Tau protein composition neurofibrillary tangles.
The incidence of Alzheimer's disease are subject to many factors,including environmental factors,poor dietary habits,drug types,such as poisoning.Chronic alcoholism lead to brain APP and Aβprotein in order to trigger an increase in the incidence of Alzheimer's disease,is still a lack of systematic theoretical and experimental studies.
Neuron-specific enolase(NSE) is a sign of nerve cell protein,the existence of specificity in neurons and neuroendocrine cells in the brain of all soluble protein accounted for 1.5%of neuronal damage or NSE into the blood after necrosis.
In this study,mice in the establishment of animal models of chronic alcoholism, based on the application of histopathological and immunohistochemical techniques, research in mice after chronic alcoholism and brain nerve cells in the expression of APP and Aβ,and the application of SPSS statistical software of the observed experimental results were statistically analyzed to explore neurons in chronic alcoholism APP and Aβ,as well as the content changes in the incidence of Alzheimer's disease and the causal link between them.
Materials and Methods
1.Establishment of animal model and packet
42 9-week-old Kunming mice were randomly divided into 30-day group,the 60-day groups was divided into 10%alcohol group,20%alcohol group and the control group,of which five of the control group,10%ethanol group 8,and 20%alcohol group 8.
The experimental group will be anhydrous ethanol(HPLC pure) in drinking water to match the concentration of 10%,20%alcohol solution as the drinking water of mice in the control group used in place of drinking water.After the end of the experiment, animals were killed directly decapitation,rapid extraction of mouse brain tissue,4% paraformaldehyde/PBS pH7.4 fixed,paraffin-embedded,producer.
2.Methods
(1) HE staining to observe the application of the morphological characteristics of organizations.
(2) the use of Nissl staining of nerve cells circumstances Nissl bodies.
(3) Application of neuron-specific enolase staining patterns characteristic of neurons.
(4) immunohistochemical staining APP and Aβin the brain tissues.
(5) the use of Motic Images Advanced 3.2 image analysis system to calculate immunohistochemical staining positive cells and expression of the area.
(6) the application of SPSS statistical software for statistical analysis of experimental data
Result
(1) The experimental group of mice significantly the performance of chronic poisoning,and decreased body weight.
(2) HE staining of chronic alcoholism see the mice brain edema,nerve cells in the disappearance of Nissl bodies or margination,nerve cells around the gap widened.
(3) neuron-specific enolase immunohistochemistry see increase in enzyme protein content.
(4) the control group shows that a small number of APP positive immunohistochemical staining of nerve cells,a product of the distribution of staining in the cell cytoplasm,showing brown yellow.Experimental group,APP staining positive cells markedly increased expression of different concentrations and different time periods of different intensity of positive expression.Experimental group,nerve cells showed a visible product of Aβ-positive staining,and high concentrations of prolonged treatment group the number of positive cells markedly increased expression.Using Motic Image Advanced 3.2 image analysis system to calculate the average integrated optical density,and the use of SPSS software for statistical analysis and found that the experimental group and the control group there was significant difference.
Conclusion
(1) Chronic alcoholism can cause brain cells of mice degeneration.
(2) Chronic alcoholism can cause brain cells of mice neuron-specific enolase increase in content and distribution of abnormalities.
(3) Chronic alcoholism can cause brain cells of mice produce Aβprotein,and APP protein expression was significantly enhanced.
(4) Chronic alcoholism may be induced by Alzheimer's disease,one of the relevant factors.
引文
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23Huang X,Atwood CS,Hartshorn MA,et al,The A beta peptide of Alzheimer's disease directly produces hydrogen peroxide through metal ion reduction Biochemistry 1999;38(24):7609-7616.
24Rachel L.Self,Katherine J.Smith,Patrick J.Mulholland,and Mark A.Prendergast.Ethanol Exposure and Withdrawal Sensitizes the Rat Hippocampal CA1 Pyramidal Cell Region to β-Amyloid(25-35)-Induced Cytotoxicity:NMDA Receptor Involvement.ALCOHOLISM,CLINICAL AND EXPERIMENTAL RESEARCH,November 2005,Vol.29,No.11:2063-2068.
25邓娟。吸烟、饮酒与老年痴呆关系的研究。第三军医大学博士学位论文2006年6月
26Ruitenberg A,van Swieten JC,Witteman JC,et al.Alcohol consumption and risk of dementia:the Rotterdam Study.Lancet 2002;359:281-286.
27Mukamal K J,Kuller LH,Fitzpatrick AL,et al.Prospective study of alcohol consumption and risk of dementia in older adults.JAMA.2003,289:1405-1413.
1 Presott CA,Kendler KS.Genetic and environmental contributions to alcohol abuse and dependence in a population-based sample of male twinsJ Am Ps ychiatry,1999,156:34-40.
2 荆晓明,董芸,宋文忠,等。急性酒精中毒患者血清β内啡肽水平及意义(J)。中华急诊医学杂志,2001,17(3):135-139.
3 KRIL J J,HOMEWOOD.Neuroal changes in the cerebral cortex of the rat following alcohol treatment and thiamin deficiency(J).J Neuropalthol Exp Neurol,1993,52(6):586-593.
4 Jaatinen P,Riikonen J,Riihioja P,et al.Interaction of aging and intermittent ethanol exposure on brain cytochrome oxidase activing level.Alcohol,2003,29(2):91.
5 庞治兴,张顺莲,张丽。17例慢性酒精中毒精神障碍的脑CT所见(J).中国神经精神病学杂志,1998,14(1):52.
6 Kril J J,Halliday GM,Svoboda MD,et al.The cerebral cortex is damaged in chronic alcoholicsJ.Neuroscience,1997,79(4):983.
7 赵丽,韩威,陈嘉峰,等.酒精中毒大鼠脑血管病变及相应脑组织损伤的病理观察(J).中风与神经疾病杂志,2005,22(1):51-53.
8 杜万君,郭菘,蒋佐宁.Wisconsin卡片分类测验用于酒精依赖者认知障碍的研究(J).中国心理卫生杂志,2002,16(5):296-298.
9 BOWDEN S C,MCCARTER R J.Spatial menory in alcohol dependent subjects:using a pushbutton maze to test the principle of equiavailability(J).Brain Cogn,1993,22(1):51-62.
10 赵丽,韩威,陈嘉峰,等.酒精中毒大鼠脑血.管病变及相应脑组织损伤的病理观察(J).中风与神经疾病杂志,2005,22(1):51-53.
11 Yamashita M,Yamamoto T.Wernicke ence phlopathy with symmetric pericentral involvement:MR findingsJ.J Corn pute Assist Tomogr,1995,19(2):306-308.
12 于爱红,朴常福 等.酒精中毒研究进展(J).中国康复理论与实践,2004;10(11):690-691.
13 张镛。酒精中毒与神经损害。《医学综述》。1995年第1卷第3期 112-114
14 Rachel L.Self,Katherine J.Smith,Patrick J.Mulholland,and Mark A.Prendergast.Ethanol Exposure and Withdrawal Sensitizes the Rat Hippocampal CA1 Pyramidal Cell Region to β-Amyloid(25-35)-Induced Cytotoxicity:NMDA Receptor Involvement.ALCOHOLISM,CLINICAL AND EXPERIMENTAL RESEARCH,November 2005,Vol.29,No.11:2063-2068.
15 邓娟。吸烟、饮酒与老年痴呆关系的研究。第三军医大学博士学位论文2006年6月
16 Tesco G,Koh YH,Kang EL,Cameron AN,Das S,Sena-Esteves M,Hiltunen M,Yang SH,Zhong Z,Shen Y,Simpkins JW,Tanzi RE.Depletion of GGA3 stabilizes BACE and enhances beta-secretase activity.Neuron.2007 Jun 7;54(5):721-37.
17 赵丽,韩威,陈嘉峰,曹树文,崔元日,李博。酒精中毒大鼠脑血管病变及相应脑组织损伤的病理观察。中风与神经疾病杂志2005年2月 第22卷第1期:51-54。
18 陈波,钱淑文。β-淀粉样蛋白免疫治疗阿尔茨海默型痴呆的研究进展。国外医学·老年医学分册2005年11月第26卷第6期 244-247
19 崔卫刚,陶晶,任铭新。慢性酒精中毒大鼠学习记忆及Meynert基底核胆碱能神经元的变化。四川解剖学杂志2006年第14卷第3期:14-16
20 Hemby SE,Connor JAO,Acosta G.et al.Ethanol-Induced Regulation of GABAA Subunit mRNAs in Prefrontal Fields of Cynomolgus Monkeys.Alcohol Clin Exp Res.2006,30(12):1978-1985
21 张癸荣,高登莲,曹龙光,殷明。γ-氨基丁酸抑制大鼠海马脑片可溶性淀粉样前体蛋白的释放。第二军医大学学报 2004年7月 25(6):609-611
22 Roberto M,Treistman SN,Pietrzykowski AZ,et al.Actions of Acute and ChronicEthanolonPresynapticTerminals.ALCOHOLISM:CLINICALANDEXPERIMENTALR ESEARCH.2006,30(2):222-232
23.Cha YM,Li Q,Wilson WA,et al.Sedative and GABAergic Effects of Ethanol on Male and Female Rats.Alcohol Clin Exp Res.2006,30(1):113-118
24 Ikonomidou C,Bittigau P,Ishimaru M J,et al.Ethanol-induced apoptotic neurodegeneration and fetal alcohol syndrome.Science,2000,287(5455):1056-60
25 Toso L,Poggi SH,Abebe D,et al.N-Methyl-D-aspartate subunit expression during mouse development altered by in utero alcohol exposure.Am J Obstet Gynecol,2005,193(4):1534-9
26 Davies M.The role of GABAA receptors in mediating the effects of alcohol in the central nervous system.J Psychia-try Neurosci,2003,28(4):263-274
27 Ikonomidou C,Bittigau P,Koch C,et al.Neurotransmitters and apoptosis in the developing brain.Biochem Pharmacol,2001,62(4):401-405
28 Young C,Roth KA,Klocke B J,et al.Role of caspase-3 in ethanol-induced developmental neurodegeneration.Neurobiol Dis,2005,20(2):608-614
29 Heaton MB,Paiva M,Madorsky I,et al.Effect of bax deletion on ethanol sensitivity in the neonatal rat cerebellum.J Neurobiol,2006,66(1):95-101
30 Siler-Marsiglio KI,Madorsky I,Pan Q,et al.Effects of acute ethanol exposure on regulatory mechanisms of Bcl-2-associated apoptosis promoter,Bad,in neonatal rat cerebellum:differential effects during vulnerable and resistant developmental periods.Alcohol Clin Exp Res,2006,30
2叶兰仙,王有德,刘建斌.慢性酒精中毒性精神障碍70例临床分析.中国行为医学科学,2002,8(2):225-226
3Alvarez AR,Goday JA,,Mullendorff K,et al.Wnt-3a overcomes beta-amyloid toxicity in rat hippocampal neurons.Exp Cell Res,2004,297(1):186-96
4Anderton BH Dayanandan R Killiek R,et al Does dysregulation of the notch and wingless Wnt pathway underlie the pathogenesis of Alzhermer's disease Mol Med Today 2000;6(2):54-9
5Reyes AE,Chacon MA,Dinamarca MC,et al.Acetylcholinesterase-Abeta complexes are more toxic than Abeta fibrils in rat hippocampus:effect on rat beta-amyloid aggregation,laminin expression,reactive astrocytosis,and neuronal cell loss.Am J Pathol 2004;164(6):2163-74.
6Dennis R.Sparta,Angela M.Sparrow,Emily G.Lowery,et al Blockade of the Corticotropin Releasing Factor Type Receptor Attenuates Elevated Ethanol Drinking Associated With Drinking in the Dark Procedures Clinical and Experimental Research 2008;32(2):269-265
7Eileen M.Moore,Kristen M.Serio,Karen J.Goldfarb,et al.GABAergic modulation of binge-like ethanol intake in C57BL/6J mice Science Direct Biochemistry and Behavior 88(2007) 105-113
8高霞.慢性酒精中毒神经系统损害分析.2007,34(20):3847-3852.
9LambertMP,Barlow AK,Chromy BA,et al Diffusible,nonfibrillar ligands derived from Abetal-42 are potent central nervous system neurotoxins(J).Proc Natl Acad Sci USA,1998,95:6448-6453.
10Tesco G,Koh YH,Kang EL,Cameron AN,Das S,Sena-Esteves M,Hiltunen M,Yang SH,Zhong Z,Shen Y,Simpkins JW,Tanzi RE.Depletion of GGA3 stabilizes BACE and enhances beta-secretase activity.Neuron.2007 Jun 7;54(5):721-37.
11赵丽,韩威,陈嘉峰,曹树文,崔元日,李博。酒精中毒大鼠脑血管病变及相应脑组织损伤的病理观察。中风与神经疾病杂志2005年2月第22卷第1期:51-54。
12陈波,钱淑文。β-淀粉样蛋白免疫治疗阿尔茨海默型痴呆的研究进展。国外医学·老年医学分册2005年11月第26卷第6期244-247
13崔卫刚,陶晶,任铭新。慢性酒精中毒大鼠学习记忆及Meynert基底核胆碱能神经元的变化。四川解剖学杂志2006年第14卷第3期:14-16
14Hemby SE,Connor JAO,Acosta G,et al.Ethanol-Induced Regulation of GABAASubunit mRNAs in Prefrontal Fields of Cynomolgus Monkeys.Alcohol Clin Exp Res.2006,30(12):1978-1985
15张癸荣,高登莲,曹龙光,殷明。γ-氨基丁酸抑制大鼠海马脑片可溶性淀粉样前体蛋白的释放。第二军医大学学报2004年7月25(6):60-61
16吕良、钟镇国.阿尔茨海默病淀粉样β蛋白诱导神经元凋亡的研究进展.2007,5(6):517-519.
17Pereira C,Ferreiro E,Cardoso SM,et al.Cell degeneration induced by amyloid-beta peptides:Implications for Alzheiler's disease(J).Jmol Neur Osci;2004,23(1-2):97-104.
18Kawasumi M,Hashimoto Y,Chiba T,et al.Molecular mechanisms for neuronal cell death by Alzheiler's amyloid precusor protein-relevant insults(M).Neurosignals,2002:236-250.
19Ingram VM.The role of Alzheimer A beta-peptides in ion transport across cell membranes(J)Subcell Biochem,2005,38:339-349.
20Ueda K,Shinohara S,Yagami T,et al.Amyloid beta protein potentiates Ca~(2+) influx through L-type voltage-sensition Ca~(2+) channels:a possible involvement of free radicals.J Neurochem 1997:68(1):265-271.
21Fiorucci L,Ascoli F.Mast cell tryptase,a still enigmatic enzyme.Cell Mol Life Sci.2004;61(11):1278-1295.
22许志强,周华东,蒋晓江.淀粉样β蛋白沉淀及其神经毒性与阿尔茨海默病.中国临床康复.2006,10(14):138-140.
23Huang X,Atwood CS,Hartshorn MA,et al,The A beta peptide of Alzheimer's disease directly produces hydrogen peroxide through metal ion reduction Biochemistry 1999;38(24):7609-7616.
24Rachel L.Self,Katherine J.Smith,Patrick J.Mulholland,and Mark A.Prendergast.Ethanol Exposure and Withdrawal Sensitizes the Rat Hippocampal CA1 Pyramidal Cell Region to β-Amyloid(25-35)-Induced Cytotoxicity:NMDA Receptor Involvement.ALCOHOLISM,CLINICAL AND EXPERIMENTAL RESEARCH,November 2005,Vol.29,No.11:2063-2068.
25邓娟。吸烟、饮酒与老年痴呆关系的研究。第三军医大学博士学位论文2006年6月
26Ruitenberg A,van Swieten JC,Witteman JC,et al.Alcohol consumption and risk of dementia:the Rotterdam Study.Lancet 2002;359:281-286.
27Mukamal K J,Kuller LH,Fitzpatrick AL,et al.Prospective study of alcohol consumption and risk of dementia in older adults.JAMA.2003,289:1405-1413.
1 Presott CA,Kendler KS.Genetic and environmental contributions to alcohol abuse and dependence in a population-based sample of male twinsJ Am Ps ychiatry,1999,156:34-40.
2 荆晓明,董芸,宋文忠,等。急性酒精中毒患者血清β内啡肽水平及意义(J)。中华急诊医学杂志,2001,17(3):135-139.
3 KRIL J J,HOMEWOOD.Neuroal changes in the cerebral cortex of the rat following alcohol treatment and thiamin deficiency(J).J Neuropalthol Exp Neurol,1993,52(6):586-593.
4 Jaatinen P,Riikonen J,Riihioja P,et al.Interaction of aging and intermittent ethanol exposure on brain cytochrome oxidase activing level.Alcohol,2003,29(2):91.
5 庞治兴,张顺莲,张丽。17例慢性酒精中毒精神障碍的脑CT所见(J).中国神经精神病学杂志,1998,14(1):52.
6 Kril J J,Halliday GM,Svoboda MD,et al.The cerebral cortex is damaged in chronic alcoholicsJ.Neuroscience,1997,79(4):983.
7 赵丽,韩威,陈嘉峰,等.酒精中毒大鼠脑血管病变及相应脑组织损伤的病理观察(J).中风与神经疾病杂志,2005,22(1):51-53.
8 杜万君,郭菘,蒋佐宁.Wisconsin卡片分类测验用于酒精依赖者认知障碍的研究(J).中国心理卫生杂志,2002,16(5):296-298.
9 BOWDEN S C,MCCARTER R J.Spatial menory in alcohol dependent subjects:using a pushbutton maze to test the principle of equiavailability(J).Brain Cogn,1993,22(1):51-62.
10 赵丽,韩威,陈嘉峰,等.酒精中毒大鼠脑血.管病变及相应脑组织损伤的病理观察(J).中风与神经疾病杂志,2005,22(1):51-53.
11 Yamashita M,Yamamoto T.Wernicke ence phlopathy with symmetric pericentral involvement:MR findingsJ.J Corn pute Assist Tomogr,1995,19(2):306-308.
12 于爱红,朴常福 等.酒精中毒研究进展(J).中国康复理论与实践,2004;10(11):690-691.
13 张镛。酒精中毒与神经损害。《医学综述》。1995年第1卷第3期 112-114
14 Rachel L.Self,Katherine J.Smith,Patrick J.Mulholland,and Mark A.Prendergast.Ethanol Exposure and Withdrawal Sensitizes the Rat Hippocampal CA1 Pyramidal Cell Region to β-Amyloid(25-35)-Induced Cytotoxicity:NMDA Receptor Involvement.ALCOHOLISM,CLINICAL AND EXPERIMENTAL RESEARCH,November 2005,Vol.29,No.11:2063-2068.
15 邓娟。吸烟、饮酒与老年痴呆关系的研究。第三军医大学博士学位论文2006年6月
16 Tesco G,Koh YH,Kang EL,Cameron AN,Das S,Sena-Esteves M,Hiltunen M,Yang SH,Zhong Z,Shen Y,Simpkins JW,Tanzi RE.Depletion of GGA3 stabilizes BACE and enhances beta-secretase activity.Neuron.2007 Jun 7;54(5):721-37.
17 赵丽,韩威,陈嘉峰,曹树文,崔元日,李博。酒精中毒大鼠脑血管病变及相应脑组织损伤的病理观察。中风与神经疾病杂志2005年2月 第22卷第1期:51-54。
18 陈波,钱淑文。β-淀粉样蛋白免疫治疗阿尔茨海默型痴呆的研究进展。国外医学·老年医学分册2005年11月第26卷第6期 244-247
19 崔卫刚,陶晶,任铭新。慢性酒精中毒大鼠学习记忆及Meynert基底核胆碱能神经元的变化。四川解剖学杂志2006年第14卷第3期:14-16
20 Hemby SE,Connor JAO,Acosta G.et al.Ethanol-Induced Regulation of GABAA Subunit mRNAs in Prefrontal Fields of Cynomolgus Monkeys.Alcohol Clin Exp Res.2006,30(12):1978-1985
21 张癸荣,高登莲,曹龙光,殷明。γ-氨基丁酸抑制大鼠海马脑片可溶性淀粉样前体蛋白的释放。第二军医大学学报 2004年7月 25(6):609-611
22 Roberto M,Treistman SN,Pietrzykowski AZ,et al.Actions of Acute and ChronicEthanolonPresynapticTerminals.ALCOHOLISM:CLINICALANDEXPERIMENTALR ESEARCH.2006,30(2):222-232
23.Cha YM,Li Q,Wilson WA,et al.Sedative and GABAergic Effects of Ethanol on Male and Female Rats.Alcohol Clin Exp Res.2006,30(1):113-118
24 Ikonomidou C,Bittigau P,Ishimaru M J,et al.Ethanol-induced apoptotic neurodegeneration and fetal alcohol syndrome.Science,2000,287(5455):1056-60
25 Toso L,Poggi SH,Abebe D,et al.N-Methyl-D-aspartate subunit expression during mouse development altered by in utero alcohol exposure.Am J Obstet Gynecol,2005,193(4):1534-9
26 Davies M.The role of GABAA receptors in mediating the effects of alcohol in the central nervous system.J Psychia-try Neurosci,2003,28(4):263-274
27 Ikonomidou C,Bittigau P,Koch C,et al.Neurotransmitters and apoptosis in the developing brain.Biochem Pharmacol,2001,62(4):401-405
28 Young C,Roth KA,Klocke B J,et al.Role of caspase-3 in ethanol-induced developmental neurodegeneration.Neurobiol Dis,2005,20(2):608-614
29 Heaton MB,Paiva M,Madorsky I,et al.Effect of bax deletion on ethanol sensitivity in the neonatal rat cerebellum.J Neurobiol,2006,66(1):95-101
30 Siler-Marsiglio KI,Madorsky I,Pan Q,et al.Effects of acute ethanol exposure on regulatory mechanisms of Bcl-2-associated apoptosis promoter,Bad,in neonatal rat cerebellum:differential effects during vulnerable and resistant developmental periods.Alcohol Clin Exp Res,2006,30