丁香酚吸嗅对VD模型大鼠学习记忆功能改善和海马区谷氨酸能神经元的影响

发布时间：2018-05-10 20:57

本文选题：丁香酚 + 血管性痴呆　；参考：《安徽医科大学》2012年硕士论文

【摘要】：目的本实验采用改良的2VO法建立慢性脑缺血VD模型，观察丁香酚吸嗅后VD模型大鼠学习障碍记忆改善的情况；并使用免疫组化法检测丁香酚吸嗅后大鼠海马组织中Glu能神经元的变化情况，，初步探讨其作用机制。方法 1.建立VD大鼠模型：用改良的2VO法造模，即先结扎右侧颈总动脉，1周后用同样的方法结扎左侧颈总动脉。 2.丁香酚对VD模型大鼠学习障碍记忆改善及机制：成年雄性SD大鼠45只随机分成3组：（1）VD模型组（n=18）：用改良的2VO方法造模，①在造模完成后7d、30d、60d时，用Morris水迷宫测定大鼠的空间学习记忆能力，记录定位航行实验中大鼠到达安全平台的时间（即为逃避潜伏期）和空间探索实验中的大鼠穿越原平台的次数（即为穿台次数）为实验指标，时间都以120s为限。②在行为学实验结束后，大鼠断头取脑，制作石蜡切片，HE染色观察大鼠海马区病理形态学变化，应用免疫组化法测定大鼠海马区的Glu能神经元的变化情况。（2）实验组（n=17）:手术的操作步骤同模型组，在造模完成后3d后予以1%浓度的丁香酚吸嗅，上、下午各1次，每次30min，同时行与模型组相同的操作步骤。（3）空白组（n=10）：不做任何手术处理，余实验步骤同模型组。结果 1.Morris水迷宫实验中，术后7d时，模型组、实验组、空白组大鼠的逃避潜伏期、穿台次数之间差异均无统计学意义（P0.05）。 2.术后30d时，实验组大鼠逃避潜伏期显著短于模型组，差异有统计学意义（P0.05）；穿台次数显著多于模型组，差异有统计学意义（P0.05）。模型组大鼠逃避潜伏期显著长于空白组,差异有统计学意义（P0.05）；穿台次数显著少于空白组.差异有统计学意义（P0.05）。 3.术后60d时，实验组大鼠的逃避潜伏期显著短于模型组（P0.01），穿台次数显著多于模型组（P0.01）。模型组大鼠的逃避潜伏期显著长于空白组,差异有显著统计学意义（P0.01）；穿台次数显著少于空白组,差异有显著统计学意义（P0.01）。 4.模型组大鼠在术后7d、30d和60d时，逃避潜伏期、穿台次数前后相比，差异有统计学意义（P0.05）；实验组大鼠在术后7d、30d和60d时，逃避潜伏期、穿台次数前后相比，差异有统计学意义（P0.05）。 5.免疫组化结果中，模型组大鼠海马区Glu阳性神经元的数量较空白组大鼠显著减少（P0.01），实验组大鼠脑组织海马区Glu阳性神经元的数量较模型组大鼠显著增加（P0.01）。结论 1.改良的2VO法建立大鼠VD模型，对大鼠脑组织损伤较小，且会出现明显的学习记忆障碍，符合VD临床的特点。 2.在脑缺血后的60d内，随着缺血时间的延长，大鼠学习记忆障碍会越严重。 3.丁香酚吸嗅后，VD模型大鼠的学习记忆障碍可明显改善。 4.丁香酚改善VD模型大鼠的学习记忆能力与抑制海马区Glu能神经元的下降有关。
[Abstract]:Purpose The VD model of chronic cerebral ischemia was established by modified 2VO method to observe the improvement of learning disability and memory in VD rats after eugenol sniffing. The changes of Glu neurons in rat hippocampal tissue after eugenol sniffing were detected by immunohistochemical method. Method 1. The model of VD rats was established by modified 2VO method. The right common carotid artery was ligated for 1 week and then left common carotid artery was ligated in the same way. 2. Effects of eugenol on learning impairment and memory in VD rats: Forty-five adult male SD rats were randomly divided into 3 groups: In the VD model group, the modified 2VO method was used to model the rats. The spatial learning and memory abilities of the rats were measured by Morris water maze at 30 days and 60 days after the completion of the model. The time to reach the safe platform (i.e. escape latency) and the number of times that the rats in the space exploration experiment crossed the original platform (i.e. the number of platform piercing) were recorded as the experimental indicators. The time limit was 120s. 2. After the behavioral experiment was finished, the rat head was cut off and his staining was made to observe the pathomorphological changes of the hippocampus, and the changes of Glu neurons in the hippocampus were measured by immunohistochemical method. (2) in the experimental group, the procedure of operation was the same as that in the model group, and 1% eugenol was given to the model group 3 days after the completion of the model, once in the afternoon, 30 minutes each time, at the same time as in the model group. Blank group: no surgical treatment, the rest of the experimental steps are the same as the model group. Result In the 1.Morris water maze test, there was no significant difference in the escape latency and the number of perforations between the model group, the experimental group and the blank group on the 7th day after operation (P 0.05). 2. At 30 days after operation, the escape latency of the experimental group was significantly shorter than that of the model group, and the difference was statistically significant (P 0.05), and the number of perforations was significantly higher than that of the model group (P 0.05). The escape latency of the model group was significantly longer than that of the blank group, the difference was statistically significant (P 0.05), and the number of platform piercing was significantly lower than that of the blank group. The difference was statistically significant (P 0.05). 3. At 60 days after operation, the escape latency of the experimental group was significantly shorter than that of the model group (P 0.01), and the number of perforations was significantly higher than that of the model group (P 0.01). The escape latency of the model group was significantly longer than that of the blank group, the difference was statistically significant (P 0.01), and the frequency of platform penetration was significantly lower than that of the blank group (P 0.01). 4. In the model group, there was a significant difference in the escape latency and the times of platform penetration between 30 and 60 days after the operation, while in the experimental group, there was a significant difference in the escape latency between 30 and 60 days after operation, and the difference between the two groups before and after the operation was statistically significant (P 0.05). 5. In the immunohistochemical results, the number of Glu positive neurons in the hippocampus of the model group was significantly lower than that of the control group, and the number of Glu positive neurons in the hippocampus of the experimental group was significantly higher than that of the model group. Conclusion 1. The modified 2VO method was used to establish VD model in rats. The damage to brain tissue was small, and obvious learning and memory disorder appeared, which was in line with the clinical characteristics of VD. 2. Within 60 days after cerebral ischemia, the learning and memory impairment would become more serious with the prolongation of ischemic time. 3. The learning and memory impairment of VD rats after eugenol sniffing can be improved significantly. 4. The effect of eugenol on learning and memory in VD rats was related to the inhibition of the decrease of Glu neurons in hippocampus.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R285.5;R749.1

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