丁香酚通过嗅觉通路对自闭症鼠学习记忆及海马区5羟色胺表达的影响

发布时间：2018-01-06 03:06

本文关键词：丁香酚通过嗅觉通路对自闭症鼠学习记忆及海马区5羟色胺表达的影响　出处：《安徽医科大学》2015年硕士论文　论文类型：学位论文

更多相关文章： 自闭症 丁香酚 嗅觉通路 5 羟色胺

【摘要】：目的本研究采用母鼠孕后12.5天单次腹腔注射丙戊酸钠600 mg/kg制备自闭症动物模型,观察丁香酚精油通过嗅觉通路干预对自闭症模型鼠学习记忆功能的改善情况;免疫组织化学染色法检测大鼠海马区5羟色胺(5-hydroxy tryptamine,5-HT)表达水平变化,初步探讨其可能的作用机制。方法1.丙戊酸钠诱导制备自闭症鼠模型清洁级SD(Sprague-Dawley)大鼠于受孕后第12.5天给予单次丙戊酸钠600 mg/kg腹腔注射制备自闭症模型,对照组给予等量生理盐水。自闭症模型组孕鼠所产幼鼠记录为丙戊酸钠组(VPA组);对照组孕鼠所产幼鼠为生理盐水组(NS组)。将幼鼠出生后当天记录为生后第一天(PN1),分别记录VPA组、NS组幼鼠发育学及行为学情况。2.丁香酚精油通过嗅觉通路对自闭症鼠学习记忆影响的实验研究:选择成功造模的P35 VPA组鼠20只,随机分为模型吸嗅组(VPA-E,n=10)和模型对照组(VPA-C,n=10),其中NS组仔鼠作为正常对照组。VPA-E组幼鼠予以1/1000的浓度稀释后丁香酚精油吸嗅,每天3次,每次30 min,共持续4周,对照组以蒸馏水代替精油。利用Morris水迷宫测定各组大鼠学习记忆能力,以定位航行实验中大鼠找到平台的时间(即逃避潜伏期)和空间探索实验中大鼠穿越平台的次数(即穿台次数)为实验的检测指标。记录丁香酚精油嗅觉吸入干预后自闭症鼠学习记忆有无改善;行为学结束后处死大鼠取脑组织制备石蜡切片,免疫组化染色法观察海马区5-HT表达水平的变化情况。结果(1)与NS组相比,VPA组幼鼠显示出生长发育的迟缓,具体表现为P7、P14、P21、P56时体重减轻,PN13-15天睁眼情况评分较低,PN7-10天方向趋向性较差,PN8、PN12时游泳表现评分较低,P9-11嗅觉辨别能力发育延迟,以上差异均具有统计学意义(P0.05);行为学方面可见VPA组幼鼠表现为社交行为次数减少及行为累积持续时间的缩短,差异具有统计学意义(P0.05);探索行为减少及重复理毛持续时间明显增多(P0.05);Morris水迷宫实验中,VPA组幼鼠表现为逃避潜伏期明显延长,差异有统计学意义(P0.05),穿越平台次数减少,差异有统计学意义(P0.05)。(2)丁香酚精油干预4周后,VPA-C组的逃避潜伏期显著长于NS组,穿越平台次数少于NS组,差异有显著统计学意义(P0.05);VPA-E组鼠逃避潜伏期短于VPA-C组、穿越平台次数多于VPA-C组,差异有统计学意义(P0.05);VPA-E组与NS组相比,逃避潜伏期及穿越平台数差异无显著性。(3)海马区免疫组化染色结果,VPA组海马区5-HT反应阳性神经元的数量较NS组减少,染色较浅,VPA-E组脑组织海马区5-HT阳性神经元的数量较VPA-C增多,染色加深。结论(1)VAP组幼鼠表现为生长发育迟缓、社交次数减少及持续时间缩短、重复行为、探索活动减少和学习记忆障碍等,VPA自闭症模型具有较好的表面效度及良好的重复性。(2)丁香酚精油经嗅觉通路对自闭症大鼠学习记忆障碍具有改善的作用。(3)丁香酚精油能够改善VPA自闭症大鼠学习记忆功能,其作用机制可能为增加海马区5-HT单胺类神经递质的表达。
[Abstract]:The purpose of this study used during pregnancy and 12.5 days after a single intraperitoneal injection of sodium valproate 600 mg/kg preparation of the autism animal model, observe the oil eugenol through olfactory pathway intervention on autism model rats learning and memory function to improve the situation; immunohistochemical staining in rat hippocampus sea (5-hydroxy tryptamine, 5-HT 5 HT) expression. To explore its possible mechanism. Methods 1. preparation of VPA induced autism model rats of clean grade SD (Sprague-Dawley) in rats after conception 12.5 days with a single intraperitoneal injection of sodium valproate 600 mg/kg preparation of the autism model, the control group received normal saline. The autism model group of pregnant rats by birth records sodium valproate group (VPA group); the control group of pregnant rats produced in the normal saline group (NS group). The postnatal day record for the first day after birth (PN1), VPA group respectively, NS of rats in group development theory and behavior of.2. eugenol oil through olfactory pathway on Experimental Study on the impact of learning and memory in rats with autism: successful models of P35 group VPA 20 rats were randomly divided into model group (VPA-E, n=10 sniffing) and model group (VPA-C, n=10), the NS group rats as normal control group and.VPA-E group were diluted to 1/1000 after oil eugenol sniffing, 3 times a day, 30 min each time, lasting 4 weeks, the control group with distilled water instead of oil. The determination of learning and memory abilities of rats by Morris water maze, the rats positioning navigation experiments to find the platform time (i.e. escape latency) and the number of rats through the platform of space exploration experiments (i.e. in Taiwan Times) were detected. Experimental records of eugenol oil olfactory inhalation after the intervention of autism rats learning and memory behavior were improved; after the rats were sacrificed to get the brain Preparation of tissue paraffin section, immunohistochemical staining was used to observe the changes of the level of expression of 5-HT in hippocampus. Results (1) compared with NS group, VPA group rats showed growth retardation, the specific performance of P7, P14, P21, P56 weight, PN13-15 score lower opening day, PN7-10 days the orientation is poor, PN8, PN12 swimming performance score lower, P9-11 olfactory discrimination ability of developmental delay, the above differences were statistically significant (P0.05); behavior visible in VPA rats were manifested as social behavior and reduce the number of accumulated duration shortened, the difference was statistically significant (P0.05); reduced exploratory behavior repeat and grooming duration was significantly increased (P0.05); Morris water maze test, VPA rats showed escape latency was prolonged and the difference was statistically significant (P0.05), cross platform decreased, the difference was statistically significant (P0. 05). (2) after 4 weeks of eugenol oil group VPA-C intervention, the escape latency was significantly longer than that of NS group, the number of crossing platform less than NS group, the difference was statistically significant (P0.05); group VPA-E, the escape latency was shorter than that in group VPA-C, the number of cross platform more than VPA-C group, the difference was statistically significant (P0.05); compared with VPA-E group and NS group, the escape latency and cross platform number had no significant difference. (3) the results of immunohistochemical staining in the hippocampus, the number of VPA 5-HT in hippocampus neurons of the group decreased compared with group NS, lighter staining, the number of positive neurons of 5-HT in brain tissue of VPA-E group was increased VPA-C staining deepen. Conclusion (1) VAP group rats showed growth retardation, reducing the number of social and shorter duration, repetitive behaviors, exploration activity decreased and the impairment of learning and memory, VPA autism model has better surface validity and good repeatability (2) of eugenol. Essential oil can improve the learning and memory impairment of autistic rats through olfactory pathway. (3) eugenol essential oil can improve learning and memory function of VPA rats with ASD, and its possible mechanism is to increase the expression of 5-HT monoamine neurotransmitters 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】：2015
【分类号】：R965

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