迷迭香吸嗅对血管性痴呆大鼠学习记忆及海马内5-HT和GABA含量的影响

发布时间：2018-05-01 15:30

本文选题：迷迭香 + 血管性痴呆　；参考：《安徽医科大学》2013年硕士论文

【摘要】：目的本实验采用改良的双侧颈总动脉结扎法建立血管性痴呆大鼠模型，观察迷迭香吸嗅后血管性痴呆大鼠学习记忆功能的改善情况；并通过免疫组化法检测迷迭香吸嗅后实验大鼠海马内5-HT和GABA含量的变化情况，初步探讨迷迭香吸入的作用机制及临床应用价值。方法 1. VD大鼠模型制备：将健康的SD大鼠用改良的2VO法造成VD模型，即先结扎左侧颈总动脉，1周后用同样的方法结扎右侧颈总动脉。 2.模型大鼠水迷宫筛查：手术后4周时，初步筛选并存活的模型大鼠16只，正常大鼠和假手术大鼠各8只。通过Morris水迷宫测定24只大鼠的空间学习记忆成绩，记录定位航行实验中大鼠找到安全平台的时间（即逃避潜伏期）和空间探索实验中大鼠穿越原平台所在位置的次数（即穿台次数）为实验指标，时间均以120s为限。 3.分组及给药：将造模成功的大鼠随机分为模型组和迷迭香组，另设正常组和假手术组对照。（1）迷迭香组（n=8）:每天上、下午分别予以1%浓度的迷迭香吸嗅，每次30min；（2）VD模型组（n=8）：使用蒸馏水替迷迭香精油，其他同迷迭香组；（3）假手术组（n=8）：假手术组分离出2侧颈总动脉但不予结扎，余实验步骤同VD模型组；（4）正常组（n=8）：不做任何手术处理，余实验步骤同模型组。 4.神经行为学和形态学观察①在用药后5周时，再次使用Morris水迷宫测定大鼠的空间学习记忆成绩，水迷宫实验的方法同前。②在神经行为学实验结束后，将全部大鼠断头取脑，制作石蜡切片，进行HE染色观察大鼠海马区的病理形态学变化，再用免疫组化法观察大鼠海马区5-HT和GABA含量的变化情况。结果 1.术后4周时，手术组的逃避潜伏期明显长于正常组，差异有统计学意义(P0.05)；穿台次数显著少于正常组,差异有显著统计学意义（P0.05）；假手术组和正常组相比，潜伏期与穿台次数均无明显差异（P＞0.05）。 2.用药5周时，，迷迭香吸嗅组的逃避潜伏期短于模型组，差异有统计学意义(P0.05)；穿台次数显著多于模型组（P0.05）。模型组大鼠的逃避潜伏期明显长于正常组,差异有显著统计学意义（P0.05）；穿台次数显著少于正常组,差异有显著统计学意义（P0.05）。 3.免疫组化结果中，模型组的5-HT和GABA的阳性神经元较正常组明显减少（P0.05），与模型组比较，迷迭香吸嗅组的阳性神经元数明显增多（P0.05）。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.迷迭香吸嗅对VD大鼠的学习记忆障碍有明显的改善作用。 3.迷迭香改善VD模型大鼠的学习记忆能力可能与增加海马内5-HT和GABA的含量有关。
[Abstract]:Purpose An improved bilateral common carotid artery ligation method was used to establish vascular dementia rat model and to observe the improvement of learning and memory function of vascular dementia rats after rosemary sniffing. The changes of 5-HT and GABA in hippocampus of rosemary were detected by immunohistochemical method, and the mechanism of rosemary inhalation and its clinical value were discussed. Method 1. Establishment of VD rat model: VD model was established by modified 2VO method in healthy SD rats. The left common carotid artery was ligated for 1 week and then the right common carotid artery was ligated in the same way. 2. Water maze screening in model rats: at 4 weeks after operation, 16 model rats were preliminarily screened and survived, 8 normal rats and 8 sham-operated rats. The spatial learning and memory scores of 24 rats were measured by Morris water maze. The time of finding the safe platform (escape latency) and the times of passing through the position of the original platform (i.e. the number of platform penetration) in the space exploration experiment were recorded as the experimental indexes, and the time limit was 120 s. 3. Group and administration: the rats were randomly divided into model group and rosemary group, and were divided into normal group and sham operation group. Rosemary group (1) rosemary group was given 1% rosemary sniffing for 30 minutes each day and afternoon. Rosemary essential oil was replaced by distilled water and other rosemary groups were treated with distilled water. (3) sham-operation group: two common carotid arteries were isolated but not ligated in sham-operation group, and the remaining experimental steps were the same as VD model group. Normal group: no surgical treatment, the rest of the experimental steps are the same as the model group. 4. Neurobehavioral and morphological observation 1 at 5 weeks after treatment, the spatial learning and memory scores of rats were determined by Morris water maze again. The method of water maze test was the same as that of the previous 2. 2. After the end of neurobehavioral experiment, all the rats were cut off their heads and their brains were removed. The histopathologic changes of hippocampus were observed by HE staining and the changes of 5-HT and GABA in hippocampus were observed by immunohistochemical method. Result 1. At 4 weeks after operation, the escape latency in the operation group was significantly longer than that in the normal group, the difference was statistically significant (P 0.05), and the number of platform piercing was significantly lower than that in the normal group (P 0.05). There was no significant difference between the incubation period and the times of platform penetration (P > 0.05). 2. At 5 weeks, the escape latency of rosemary sniffing group was shorter than that of model group, the difference was statistically significant (P 0.05), and the number of puncture was significantly higher than that of model group (P 0.05). The escape latency of the model group was significantly longer than that of the normal group, and the difference was statistically significant (P 0.05), and the frequency of platform penetration was significantly lower than that of the normal group (P 0.05). 3. The positive neurons of 5-HT and GABA in the model group were significantly lower than those in the normal group, and the number of positive neurons in rosemary sniffing group was significantly higher than that in 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. Rosemary sniffing can significantly improve learning and memory impairment in VD rats. 3. Rosemary could improve the learning and memory ability of VD rats by increasing the contents of 5-HT and GABA 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】：2013
【分类号】：R749.13

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