大鼠黑质多巴胺能神经元衰老性变化的多项参数测量  

Aging changes of dopaminergic neurons in substantia nigra of rats:Analysis of multiple parameters

作　　者：余资江[1] 康朝胜[1] 余德立[2] 王景传[1] 许庭良[3] 姜俸蓉[3] 

机构地区：[1]贵阳医学院解剖学教研室,贵州省贵阳市550004 [2]贵阳中医学院第二附属医院眼科,贵州省贵阳市550001 [3]贵阳医学院组织胚胎学教研室,贵州省贵阳市550004

出　　处：《中国组织工程研究与临床康复》2007年第16期3108-3111,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30660189)~~

摘　　要：目的:探讨大鼠中脑黑质多巴胺能神经元的衰老性变化规律,为进一步揭示黑质病变的病因提供客观依据。方法:实验于2005-07/2006-07在贵阳医学院解剖学教研室完成。选择健康SD大鼠32只,雌雄各半,按随机数字表法分为幼年组(1～2个月龄)、青年组(4～5个月龄)、中年组(11～12个月龄)、老年组(≥24个月龄)4组,每组8只。取中脑组织常规石蜡包埋,行中脑黑质连续冠状切片,焦油紫染色、酪氨酸羟化酶免疫组织化学染色,图像分析仪测量酪氨酸羟化酶免疫阳性产物的吸光度值和酪氨酸羟化酶免疫反应阳性神经元的胞面积、体密度、数密度、圆球度。结果:32只大鼠均进入结果分析。①中脑黑质焦油紫染色形态学观察:幼年组与青年组大鼠黑质神经元胞体大,细胞排列密集,形状规则,细胞成椭圆形或锥体形,胞浆丰富,每个细胞有清晰的胞核,核仁清楚可见,尼氏体粗大而染色深,中年组和老年组大鼠神经元胞浆染色较淡,尼氏体较分散,细胞散在,数量少,可见软化灶形成。幼年组、青年组、中年组、老年组计数单位面积焦油紫染色细胞数分别为(48.00±9.10),(65.00±8.73),(20.00±4.10),(13.25±1.83)个/40倍视野,各组间比较差异有显著性意义(F=3.79,P<0.05)。②中脑黑质酪氨酸羟化酶免疫反应阳性神经元:光镜下,酪氨酸羟化酶免疫反应阳性神经元成群分布,胞浆内阳性产物以幼年组和青年组为显著,中年组和老年组胞浆内酪氨酸羟化酶阳性反应颗粒明显减少,部分神经元丧失大多角形或锥体形形状,胞体变大,排列不规则,数量减少。幼年组、青年组、中年组、老年组胞浆内酪氨酸羟化酶免疫反应阳性产物吸光度值分别为0.1993±0.0711,0.2428±0.1729,0.1978±0.0687,0.1671±0.1018,各组间比较差异有显著性意义(F=1.87,P<0.05)。中年组、老年组酪氨酸羟化酶免疫反应阳性神经元体密度、数密度低于青年组,差�AIM： To study the regularities of dopamine neurons in substantia nigra of middle brain with age increasing, and further provide objective evidence for revealing lesion of substantia nigra. METHODS： The experiment was carded out in the Department of Anatomy, Guiyang Medical University between July 2005 and July 2006. Thirty-two healthy SD rats of either gender were used in the experiment and randomized into four groups with 8 in each： juvenile group （1-2 months）, youth group （4-5 months）, middle-aged group （11-12 months） and old-aged group （≥24 months）. Routine paraffin was made in continuous coronal sections of substantia nigra for Nissl （Cresyl Violet） staining and Tyrosine hydroxylase （TH） immunohistochemical staining. The average optical density of TH immune positive product, volume density, numerical density, mean profile area, number density, average volume, sphericity of the TH-positive neurons were measured and calculated with image analyze, and the Nissl stained cells were counted. RESULTS： All the 32 rats were involved in the analysis of results.（1）Neuronal morphology： The neurons in substantia nigra of midbrain ranged regularly with oval or pyramid shape and rich kytoplasm. Each cell ,presented clear nucleus and nucleolus in rats of juvenile group and youth group. Additionally the Nisslstaining was dark. In middle-aged group and old-aged group, the neurons were dyed a little, Nissl body scattered and cell number were decreased so as to form softening lesion. There were significant differences in the 4 groups for cell count of Nissl staining [（48.00±9.10）, （65.00± 8.73）, （20.00±4.10）, （13.25±1.83） per visual field, F =3.79, P 〈 0.05].（2）TH-positive neurons of the substantia nigra of midbrain in rats： The TH-positive reactive product were principally distributed in cluster over the cytoplasm of dopaminergic neurons in substantia nigra of midbrain, especially significant in juvenile group and youth group; but the product of TH-positive

关 键 词：酪氨酸羟化酶 神经元 衰老 黑质 大鼠 

分 类 号：R744.8[医药卫生—神经病学与精神病学]