發(fā)布時間：2018-01-06 03:06

  本文關(guān)鍵詞：丁香酚通過嗅覺通路對自閉癥鼠學(xué)習(xí)記憶及海馬區(qū)5羥色胺表達(dá)的影響　出處：《安徽醫(yī)科大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 自閉癥 丁香酚 嗅覺通路 5 羥色胺

【摘要】：目的本研究采用母鼠孕后12.5天單次腹腔注射丙戊酸鈉600 mg/kg制備自閉癥動物模型,觀察丁香酚精油通過嗅覺通路干預(yù)對自閉癥模型鼠學(xué)習(xí)記憶功能的改善情況;免疫組織化學(xué)染色法檢測大鼠海馬區(qū)5羥色胺(5-hydroxy tryptamine,5-HT)表達(dá)水平變化,初步探討其可能的作用機制。方法1.丙戊酸鈉誘導(dǎo)制備自閉癥鼠模型清潔級SD(Sprague-Dawley)大鼠于受孕后第12.5天給予單次丙戊酸鈉600 mg/kg腹腔注射制備自閉癥模型,對照組給予等量生理鹽水。自閉癥模型組孕鼠所產(chǎn)幼鼠記錄為丙戊酸鈉組(VPA組);對照組孕鼠所產(chǎn)幼鼠為生理鹽水組(NS組)。將幼鼠出生后當(dāng)天記錄為生后第一天(PN1),分別記錄VPA組、NS組幼鼠發(fā)育學(xué)及行為學(xué)情況。2.丁香酚精油通過嗅覺通路對自閉癥鼠學(xué)習(xí)記憶影響的實驗研究:選擇成功造模的P35 VPA組鼠20只,隨機分為模型吸嗅組(VPA-E,n=10)和模型對照組(VPA-C,n=10),其中NS組仔鼠作為正常對照組。VPA-E組幼鼠予以1/1000的濃度稀釋后丁香酚精油吸嗅,每天3次,每次30 min,共持續(xù)4周,對照組以蒸餾水代替精油。利用Morris水迷宮測定各組大鼠學(xué)習(xí)記憶能力,以定位航行實驗中大鼠找到平臺的時間(即逃避潛伏期)和空間探索實驗中大鼠穿越平臺的次數(shù)(即穿臺次數(shù))為實驗的檢測指標(biāo)。記錄丁香酚精油嗅覺吸入干預(yù)后自閉癥鼠學(xué)習(xí)記憶有無改善;行為學(xué)結(jié)束后處死大鼠取腦組織制備石蠟切片,免疫組化染色法觀察海馬區(qū)5-HT表達(dá)水平的變化情況。結(jié)果(1)與NS組相比,VPA組幼鼠顯示出生長發(fā)育的遲緩,具體表現(xiàn)為P7、P14、P21、P56時體重減輕,PN13-15天睜眼情況評分較低,PN7-10天方向趨向性較差,PN8、PN12時游泳表現(xiàn)評分較低,P9-11嗅覺辨別能力發(fā)育延遲,以上差異均具有統(tǒng)計學(xué)意義(P0.05);行為學(xué)方面可見VPA組幼鼠表現(xiàn)為社交行為次數(shù)減少及行為累積持續(xù)時間的縮短,差異具有統(tǒng)計學(xué)意義(P0.05);探索行為減少及重復(fù)理毛持續(xù)時間明顯增多(P0.05);Morris水迷宮實驗中,VPA組幼鼠表現(xiàn)為逃避潛伏期明顯延長,差異有統(tǒng)計學(xué)意義(P0.05),穿越平臺次數(shù)減少,差異有統(tǒng)計學(xué)意義(P0.05)。(2)丁香酚精油干預(yù)4周后,VPA-C組的逃避潛伏期顯著長于NS組,穿越平臺次數(shù)少于NS組,差異有顯著統(tǒng)計學(xué)意義(P0.05);VPA-E組鼠逃避潛伏期短于VPA-C組、穿越平臺次數(shù)多于VPA-C組,差異有統(tǒng)計學(xué)意義(P0.05);VPA-E組與NS組相比,逃避潛伏期及穿越平臺數(shù)差異無顯著性。(3)海馬區(qū)免疫組化染色結(jié)果,VPA組海馬區(qū)5-HT反應(yīng)陽性神經(jīng)元的數(shù)量較NS組減少,染色較淺,VPA-E組腦組織海馬區(qū)5-HT陽性神經(jīng)元的數(shù)量較VPA-C增多,染色加深。結(jié)論(1)VAP組幼鼠表現(xiàn)為生長發(fā)育遲緩、社交次數(shù)減少及持續(xù)時間縮短、重復(fù)行為、探索活動減少和學(xué)習(xí)記憶障礙等,VPA自閉癥模型具有較好的表面效度及良好的重復(fù)性。(2)丁香酚精油經(jīng)嗅覺通路對自閉癥大鼠學(xué)習(xí)記憶障礙具有改善的作用。(3)丁香酚精油能夠改善VPA自閉癥大鼠學(xué)習(xí)記憶功能,其作用機制可能為增加海馬區(qū)5-HT單胺類神經(jīng)遞質(zhì)的表達(dá)。
[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.

【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R965

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本文編號：1386006