获得性免疫反应与海人藻酸引起的C57BL/6小鼠海马损伤的关系(英文)  

Adaptive immune response is involved in kainic acid- induced hippocampal injury in C57BL/6 mice

作　　者：祝威[1] 崔香艳[1] 祝捷[2] 

机构地区：[1]吉林大学第一医院耳鼻喉头颈科,吉林省长春市130021 [2]吉林大学第一医院神经内科

出　　处：《中国组织工程研究与临床康复》2007年第4期785-787,800,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:海人藻酸引起在啮齿类动物的海马损伤是用于研究人类神经退行性病变的优质模型。虽然许多研究已证实炎症分子和反应参与并且促进了疾病的进程,但是否获得性免疫反应,尤其是免疫活性细胞,如T细胞和B细胞是否参与了神经退行性病变尚不清楚。目的:观察B和T细胞亚型在海人藻酸引起的海马神经元退行性病变中的作用。设计:随机对照动物实验。单位:吉林大学第一医院耳鼻喉头颈科和神经内科,瑞典卡若林斯卡医学院Huddinge医院神经医学系老年科。材料:实验于2000-06/09在瑞典KarolinskaInstitute医学院神经医学系完成。选用20只的雄性C57BL/6小鼠(野生型),同样基因背景的雄性CD4(-/-),CD8(-/-),CD4/CD8(-/-)和Igh-6(-/-)基因敲除C57BL/6小鼠分别为17,19,15,14只,鼠龄5￣6周,体质量18￣20g。3只鼠龄和体质量相匹配的C57BL/6小鼠经鼻给水作为对照。试剂和仪器:海人藻酸购自Sigma,USA。双色流式细胞仪和CellQuest购自BectonDickinson,CA,USA。方法:①所有小鼠经麻醉后,按48mg/kg剂量将7.69g/L海人藻酸滴入85只小鼠双侧鼻孔中。3只对照小鼠鼻内滴入等量水。②观察小鼠临床症状,临床症状分级如下:0￣6分,0分:正常;6分:死亡。③给予海人藻酸4.0￣5.0h后,采用流式细胞术测定脾细胞表面标记的表达。④给药7d后,麻醉所有小鼠,取脑,固定,包埋。采用尼氏染色方法评估(海马)切片神经元形态,判断神经细胞和神经退行的严重程度。应用半定量系统,即病理评分判断神经细胞和神经退行性病变的严重程度:0￣6分,0分:正常;6分:重度神经元脱失(CA3区有>40%神经元脱失)。⑤多组间差异比较采用单因素方差分析,两组间差异比较采用t检验。主要观察指标:各组小鼠临床级别、海马神经病理学改变和脾细胞表面细胞分子表达。结果:小鼠85只均进入结果分析。①临床级别:所有CD4(-/-)小鼠表现出严重癫痫�BACKGROUND： Kainic acid （KA）-induced hippocampal injury in rodents is a good model for studying human neurodegenerative diseases. Although many studies have evidenced that inflammatory molecules and responses participate in and accelerated the process of disease, it is still unclear whether adaptive immune response, especially immune competent cells, such as T and B cells, is involved in the pathogenesis of neurodegenerative diseases. OBJECTIVE： To observe the roles of B and T cell subsets in KA-induced hippocampal neurodegeneration DESIGN： Randomized controlled animal tda SETTING： Department of Otolaryngology and Head, and Department of Neurology, First Hospital, Jilin University; Division of Geriatrics, Department of Neurotec, Huddinge Hospital, Karolinska Institute. MATERIALS： This trial was conducted in the Department of Neurotec, Huddinge Hospital, Karolinska Institute during June to September 2000. Twenty male C57 BL/6 mice （wide-type）, and knockout mice CD4（-/-） （n =17）, CD8（-/-）（n =19）, CD4/CD8（-/-） （n =15） and lgh-6（-/-） （n =14） of C57BL/6 background were involved in this trial. They were aged 5 to 6 weeks,weighing 18 to 20 g. Three age-and body mass-matched C57BL/6 mice received water as controls. Reagent and instruments： KA （Sigma, USA）. Bicolor flow cytometer and CellQuest （Becton Dickinson, CA, USA）. METHODS ： ① Eighty-five anesthetized mice were slowly administrated with 7.69 g/L KA by micropipette which was connected to nose of mouse at the dose of 48 mg/kg. Three control C57BL/6 mice received the same amount of water intranasally. ②Clinical symptoms of mice were monitored. Seizures were graded using a 6-point scale, 0： normal; 6： death. ③After 4 to 5 hours of administration of KA, surface immunofluorescence staining of spleen cells was measured with flow cytometer. ④After 7 days of administration of KA, all the mice were anesthetized, and their brains were harvested, then fixed and embedded. For assessment of the severity an

关 键 词：海人藻酸 获得性免疫反应 海马神经元退行性病变 

分 类 号：R392[医药卫生—免疫学]