【作者】 白晶；

【导师】 吴江；

【作者基本信息】 吉林大学 ， 神经病学， 2009， 博士

【摘要】 本研究验证经鼻—脑通路给予神经保护肽(NAP)对实验性痴呆大鼠的神经保护作用,为老年性痴呆治疗提供新的途径;进而应用蛋白质组学技术研究NAP对实验性痴呆大鼠海马蛋白质表达的影响,筛选出NAP对实验性痴呆大鼠具有神经保护作用的差异蛋白,为阐明NAP对实验性痴呆大鼠治疗作用的机制提供理论依据。本研究主要经鼻-脑通路给予神经保护肽(NAP)验证对实验性痴呆大鼠的神经保护作用。将凝聚态的Aβ1-40联合应用转移生长因子β1(TGFβ1)注入实验大鼠左侧海马区制备实验性痴呆动物模型:通过Morris水迷宫定位航行实验检测大鼠记忆能力;通过HE染色观察各组大鼠海马区的形态学改变;通过原位杂交的方法检测实验各组大鼠海马区NF-κBmRNA,caspase3mRNA表达的变化。应用蛋白组学方法筛选出NAP对实验性痴呆大鼠具有神经保护作用的差异蛋白。本研究得到如下结果:痴呆组大鼠的记忆能力明显下降,rLent/NT4-NAP组大鼠较痴呆组大鼠记忆能力明显改善;HE染色发现痴呆组大鼠左侧海马区神经细胞明显变性、坏死,而rLent/NT4-NAP组细胞损伤情况较痴呆组大鼠明显减轻;检测痴呆组大鼠左侧海马区神经细胞胞浆内NF-kBmRNA, caspase3mRNA强阳性表达,而rLent/NT4-NAP组左侧海马区神经细胞胞浆内NF-kBmRNA, caspase3mRNA表达较痴呆组显著减少。通过质谱鉴定和数据库检索,鉴定出有5种蛋白质表达水平发生变化。分别为膜联蛋白A5、谷氨酸、蛋白激酶C、白细胞粘附分子、基底细胞粘附分子。本研究证明了NAP对实验性痴呆大鼠具有神经保护作用。首次应用蛋白质组学的方法研究了NAP对实验性痴呆大鼠脑内蛋白质表达的影响,发现膜联蛋白A5、谷氨酸、蛋白激酶C三种蛋白均参与了神经退行性疾病神经元细胞的凋亡,为研究NAP治疗老年性痴呆的作用机制提供了理论依据。更多还原

【Abstract】 Alzheimer disease (AD) is a retrogression nervous system disease. Up to now,its pathogenesy and etiology is undetermined.there is not effective treat method. Gene therapy was made use of retrogression nervous system disease.It is a technology method that purpose gene import demic cell.It bring into full play biological effect. Nearly year,we construction lentivirus viral vector of fusion gene and NT4-NAP.To reseach about neuron protective effect and mechanism of Lentivirus neuroprotective peptid to experiment dementia rat and to reseach difference protein of senile dementia disease model rat hippocamp after curing of Lentivirus neuroprotective peptid and to screen relative proteinum by proteomics technology.Probation neuroprotective effect of Lentivirus neuroprotective peptid to experiment dementia rat by nose- sermion iter.In this expriment we make some rats as the senile dementia animal model whose left hippocamp being injeced aggregated Aβ1-40 and transfer growth factorβ1 (TGFβ1). The result shows that by contrast of normal group, the memory of rats in dementia group decrease obviously, degenerations and necrosis existing in a lot of neurons, the strong masculine of NF-kBmRNA, casepse3mRNA.. In this experiment the rats in NAP group show better memory and learning in water maze, decreased necrosis in neurons, weak masculine of NF-kBmRNA, casepse3mRNA. These results tell us that NAP has neuroprotective function.Beta-amyloid protein injection into hippocampus in brain lissue were applied to make experimental rat model of Alzheimer disease. Lentivirus neuroprotective peptid was given by the way of dropping nose. the change of ethology and express NF-kBmRNA, casepse3mRNA. of hippocampal neurons in Alzheimer disease rats after therapeutic lentivirus neuroprotective peptid by the water maze and hybridization in situ tecknique.To reseach difference protein of Alzheimer disease model rat hippocamp after curing of Lentivirus neuroprotective peptid and to screen relative proteinum of neuron protective effect by proteomics technology between control group, senile dementia group and Lentivirus neuroprotective peptid group.result:Lentivirus neuroprotective peptid group on the ability of learning and memory in Alzheimer disease rats and the expressing NF-kBmRNA, casepasemRNA of hippocampal neurons were significant higher than those group. There were 5 proteinum in Alzheimer disease model rat hippocamp: glutamate carboxy peptidase, mast cell protease, activated leukocyte cell adhesion molecule, basal cell adhesion molecule and annexin A5.Conclusion In this experiment, Proving the neuron protective effect of Lentivirus neuroprotective peptid to experiment dementia rat and reseached 5 proteins between the rLent/NT4-NAP group rat hippocamp after curing of Lentivirus neuroprotective peptid and control group, Alzheimer disease group rat. It is the foundation of the mechanism action on curing the senile dementia disease.更多还原