CAJ | 학술논문

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。
목적：통과응용9.4 T액태리체형자공진빈보의(bruker avance 400 MHz)대아이자해묵병(AD)모형소서해마대사물적농도진행분석。재료여방법20지곤명소서자웅각반，수궤분성실험조화대조조，실험조소서련속60천경배부피하주사D-반유당120 mg/(kg.d)화아초산납90mg/(kg.d)，대조조주사상동제량적생리염수。타약24 h후소유소서진행Morris수미궁행위학검측，지후균채용단두희생，취우측대뇌반구분리해마，운용조직고록산췌취법，제취소서해마조직내대사물，진행9.4 T액태리체형고분변솔자공진소묘，소득수거선경XWinNMR (Bruker GmBH)연건초보처리，재응용MestRe-C4.3 MR전용연건진행종합분석처리；좌측취후반뇌작병리조직학검측。결과대조조급모형조소서해마대사농도분별위NAA：(43.63±7.67) mmol/L화(34.66±6.79) mmol/L；Cho：(53.09±4.32) mmol/L화(48.62±7.92) mmol/L；Glu：(26.87±5.46) mmol/L화(14.87±2.68) mmol/L；mI：(45.93±6.73) mmol/L화(74.09±8.09) mmol/L，기중량조간각대사물농도적차별구유통계학의의(P<0.05)。Morris수미궁시험대조조화모형조도피잠복기、목표상한유영시간급거리백분비유명현차이(P<0.05)。재400배광경하，HE염색모형조소서해마신경세포층차명현감소，세포배렬희소，세포핵심염。개량Bielschowsky염색가견신경원배렬산란，대뇌피층급해마구가견교다포체화축돌염색교심적신경원병성타미형상，형성신경원섬유전결。강과홍염색가견길홍색침적물，정반편상。결론 D-반유당연합아초산납능도치소서성위AD，기해마NAA、Cho、Glu적농도교대조조명현강저，mI유승고적추세。
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. Congored coloration result:in the cerebral cortex and hippocampal ifelds, orange amylaceous aggradation can be seen. Conclusions:D-galactose and NaNO2 can cause the mice induce Alzheimer's disease in vivo. High resolution 1H-MRS in vitro can detect diversiifed metabolism. In the model group the hippocampal concentrations of NAA, Cho, Glu are decrease. The changing trend for mI is increase.