9.4T1H-MRS定量检测阿尔茨海默病模型小鼠海马代谢物浓度的研究  

Quantitative research for metabolites by 9.4 T 1H-MRS in hippocampus of Alzheimer＇s disease mice

作　　者：庞丽[1,2] 姚建莉[1] 陈耀文[3] 李海红[4] 尤克增[1] 徐志锋[1] 吴仁华[1] 

机构地区：[1]汕头大学医学院第二附属医院放射科,汕头515041 [2]北京市昌平区中西医结合医院放射科,北京102208 [3]汕头大学中心实验室,汕头515063 [4]汕头大学医学院精神卫生中心,汕头515063

出　　处：《磁共振成像》2014年第4期302-308,共7页Chinese Journal of Magnetic Resonance Imaging

基　　金：国家自然科学基金重点项目资助(编号:30930027)

摘　　要：目的通过应用9.4 T液态离体型磁共振频谱仪(bruker avance 400 MHz)对阿尔茨海默病(AD)模型小鼠海马代谢物的浓度进行分析。材料与方法 20只昆明小鼠雌雄各半,随机分成实验组和对照组,实验组小鼠连续60天颈背部皮下注射D-半乳糖120 mg/(kg.d)和亚硝酸钠90mg/(kg.d),对照组注射相同剂量的生理盐水。打药24 h后所有小鼠进行Morris水迷宫行为学检测,之后均采用断头牺牲,取右侧大脑半球分离海马,运用组织高氯酸萃取法,提取小鼠海马组织内代谢物,进行9.4 T液态离体型高分辨率磁共振扫描,所得数据线经XWinNMR(Bruker GmBH)软件初步处理,再应用MestRe-C4.3 MR专用软件进行综合分析处理;左侧取后半脑作病理组织学检测。结果对照组及模型组小鼠海马代谢浓度分别为NAA:(43.63±7.67)mmol/L和(34.66±6.79)mmol/L;Cho:(53.09±4.32)mmol/L和(48.62±7.92)mmol/L;Glu:(26.87±5.46)mmol/L和(14.87±2.68)mmol/L;mI:(45.93±6.73)mmol/L和(74.09±8.09)mmol/L,其中两组间各代谢物浓度的差别具有统计学意义(P<0.05)。Morris水迷宫试验对照组和模型组逃避潜伏期、目标象限游泳时间及距离百分比有明显差异(P<0.05)。在400倍光镜下,HE染色模型组小鼠海马神经细胞层次明显减少,细胞排列稀疏,细胞核深染。改良Bielschowsky染色可见神经元排列散乱,大脑皮层及海马区可见较多胞体和轴突染色较深的神经元并成拖尾形状,形成神经元纤维缠结。刚果红染色可见桔红色沉积物,呈斑片状。结论 D-半乳糖联合亚硝酸钠能导致小鼠成为AD,其海马NAA、Cho、Glu的浓度较对照组明显降低,mI有升高的趋势。Objective：To study Alzheimer＇s disease mice caused by the D-galactose and NaNO2, observe the alteration of hippocampal metabolites using in vitro 9.4 T high resolution magnetic resonance spectroscopy. Materials and Methods：Twenty kunming mice were randomly divided into two groups, model group and control group. Female to male ratio was 1：1. The mice in the model group were subcutaneouly injected with D-galactose 120 mg/（kg.d）, NaNO2 90 mg/（kg.d） for 60 days and the control group with oral saline. 24 hours later after Morris water maze test, all mice were sacriifced and the right hippocampus were dissected for 1H-MRS examination. Data were collected using in vitro 9.4 T high resolution magnetic resonance spectrometer. Spectra were processed using XWINNMR and MestRe-c 4.3. Compared with the left side, HE and Bielschowsky silver impregnation and congored coloration were employed to detect and conifrm the change of brain cells. Results：Good 1H-MR spectra of perchloric acid extract from hippocampus tissue of mice were obtained. The conventional metabolites were detected and assigned. Mean concentrations of metabolites in control group and model group were, NAA：（43.63±7.67） mmol/L and （34.66±6.79） mmol/L. Cho：（53.09±4.32） mmol/L and （48.62±7.92） mmol/L. Glu：（26.87±5.46） mmol/L and （14.87±2.68） mmol/L. mI：（45.93±6.73） mmol/L and （74.09±8.09） mmol/L, the differences of two groups were statistically signiifcance （P〈0.05）. Results of Morris water maze behavior examination were as： the escape lateney and the movement distance percentage and time percentage were significant difference between control group and model group （P〈0.05）. The neurons in control group were intact and arrange tightly. In the model, pyramidal neurons either presented a densely stained shrunken appearance with minimal cytoplasm or had disappeared. Bielschowsky silver impregnation, many neuroifbrillary tangles were found, and neuropil threads are stained in model group. Cong

关 键 词：阿尔茨海默病 海马 代谢 磁共振成像 动物 实验 

分 类 号：R445.2[医药卫生—影像医学与核医学] R741[医药卫生—诊断学]