大鼠脑组织中甲状腺素受体THRα1的表达与阿尔茨海默病发病机制的实验研究

发布时间：2018-04-17 18:20

本文选题：甲状腺素受体α1 + 阿尔茨海默病　；参考：《兰州大学》2012年硕士论文

【摘要】：目的：复制阿尔茨海默病动物模型,观察其大脑皮层和海马区脑组织中甲状腺素受体THR α1的表达与阿尔茨海默病发生的相关性,并从THRα1表达增加对神经细胞tau蛋白磷酸化影响的角度对其机制进行初步探讨。方法：SD大鼠腹腔注射D-半乳糖50mg·kg-1·d-1,复制阿尔茨海默病动物模型,另设正常对照组,腹腔注射等量生理盐水。连续给药6w后观察观察动物一般情况如精神状态、活动情况、皮毛等；经二次固定法固定后取全脑组织制作病理切片,进行HE染色和改良Bicschowsky染色,用以观察大脑皮层及海马组织中老年斑,神经元纤维缠结及神经元数量、体积、排列状况等形态学变化；分别用实时荧光定量PCR (Real time fluorescence quantitation PCR, FQ-PCR)法和Western blot法检测阿尔茨海默病动物与正常大鼠脑组织中THRα1、CDK-5及p35mRNA的差异性表达和pser404-tau蛋白的表达水平。结果：与正常对照组相比,阿尔茨海默病模型组动物神情呆滞，反应迟钝,撮毛等；镜下见大脑皮层及海马区神经元排列紊乱、层次减少,核呈固缩、深染状,神经元轴突染色较深呈拖尾状,形成神经元纤维缠结,尤以海马区表现更为显著。与对照组相比,模型组皮层区THRα1、CDK-5和p35mRNA表达量增加,分别为1.20±0.30、4.71±0.54、2.11±0.40,差异有统计学意义(P0.05)；模型组海马区三者的表达量分别为2.53±0.65、18.51±2.77、5.96±0.49,与对照组相比亦有统计学差异(P0.05)模型组pser404-tau蛋白呈高表达状态,与正常对照组相比P0.05。结论：阿尔茨海默病动物大脑皮层及海马区神经细胞中THRα1mRNA的表达量显著高于正常对照组大鼠,过高THRα1可提高tau蛋白磷酸化相关激酶CDK-5mRNA及其调节因子p35mRNA的表达水平,进而加剧神经细胞tau蛋白磷酸化,导致动物大脑皮层及海马区出现神经元丢失、神经纤维缠结等典型的AD形态学变化,这可能是THRα1过表达导致阿尔茨海默病发生的主要机制之一。
[Abstract]:Objective: to study the relationship between the expression of thyroxine receptor THR 伪 1 in cerebral cortex and hippocampus and the occurrence of Alzheimer's disease.The effect of THR 伪 1 expression on the phosphorylation of tau protein in nerve cells was also discussed.Methods Dgalactose 50mg kg-1 d-1 was injected intraperitoneally to make Alzheimer's disease animal model. The normal control group was set up and the same amount of normal saline was injected intraperitoneally.After 6 weeks of continuous administration, the general conditions of animals, such as mental state, activity, fur and so on, were observed, and the whole brain tissue was fixed by two fixation methods to make pathological sections for HE staining and modified Bicschowsky staining.To observe the morphological changes of senile plaques, the number, volume and arrangement of neurons in cerebral cortex and hippocampal tissue.Real time fluorescence quantitation PCR (FQ-PCR) and Western blot were used to detect the differential expression of THR 伪 1 CDK-5 and p35mRNA and the expression of pser404-tau protein in brain tissue of Alzheimer's disease and normal rats, respectively.Results: compared with the normal control group, the Alzheimer's disease model group showed dull expression, slow response, hair and so on, and the neurons of cerebral cortex and hippocampus were disordered, the layers decreased, the nucleus was pyknotic, deep stained, and so on.The axon staining of the neurons was deeply trailing, forming the neuronal fibrillary tangles, especially in the hippocampal area.Compared with the control group, the expression of THR 伪 1, CDK-5 and p35mRNA in the cortical area of the model group was increased.The expression of pser404-tau protein in the hippocampal area of the model group was 2.53 卤0.65 卤2.775 卤0.49, respectively, which was significantly higher than that in the control group (P 0.05) and the expression of pser404-tau protein in the model group was significantly higher than that in the normal control group (P 0.05). The expression of pser404-tau protein in the model group was significantly higher than that in the control group (P 0.05), and that in the hippocampal area of the model group was 2.53 卤0.65 卤2.775 卤0.49, which was also significantly higher than that in the control group (P 0.05).Conclusion: the expression of THR 伪 1mRNA in cerebral cortex and hippocampal neurons of Alzheimer's disease rats is significantly higher than that in normal control rats. Excessive THR 伪 1 can increase the expression of tau protein phosphorylated kinase CDK-5mRNA and its regulatory factor p35mRNA.In turn, the phosphorylation of tau protein in nerve cells is aggravated, resulting in the loss of neurons and neuronal tangles in the cortex and hippocampus of animals, and typical AD morphological changes, such as neurofibrillary tangles, and so on.This may be one of the main mechanisms of overexpression of THR 伪 1 leading to Alzheimer's disease.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R749.16

【相似文献】