BDNF和VEGF在慢性應(yīng)激抑郁模型小鼠神經(jīng)元損傷中的作用及其機(jī)制
發(fā)布時(shí)間：2018-07-29 15:23
【摘要】：研究目的:建立小鼠慢性不可預(yù)見性應(yīng)激抑郁模型,研究抑郁模型小鼠腦內(nèi)海馬區(qū)(DG區(qū)、CA1區(qū)、CA3區(qū))與前額葉皮層VEGF和BDNF的表達(dá)及其與抑郁癥發(fā)生的關(guān)系,探究其在抑郁癥發(fā)生中的作用及其相關(guān)分子機(jī)制。研究方法:隨機(jī)將2月齡昆明系小白鼠分為對(duì)照組和應(yīng)激組。對(duì)照組小鼠正常飼養(yǎng),應(yīng)激組小鼠采用7種不同的應(yīng)激因子(禁水、禁食、高臺(tái)、斜籠、足底電擊、冰水游泳、通宵照明)21天,建立慢性不可預(yù)見性應(yīng)激抑郁模型。實(shí)驗(yàn)一:采用曠場實(shí)驗(yàn)、懸尾實(shí)驗(yàn)、Morris水迷宮實(shí)驗(yàn)方法,檢測小鼠行為和空間學(xué)習(xí)記憶能力各項(xiàng)指標(biāo)的變化,通過免疫組化實(shí)驗(yàn)檢測各組小鼠腦內(nèi)海馬(DG區(qū)、CA1區(qū)、CA3區(qū))和前額葉皮層BDNF、VEGF及PI3K表達(dá)量的變化,以及通過HE染色,觀察海馬區(qū)和前額葉皮層神經(jīng)元的形態(tài)變化。實(shí)驗(yàn)二:采用腦立體定位技術(shù),小鼠雙側(cè)海馬分別注射生理鹽水和LY294002。C-ns組:對(duì)照組海馬內(nèi)注射生理鹽水(NS),C-ly組:對(duì)照組海馬內(nèi)注射LY294002;S-ns組:應(yīng)激組海馬內(nèi)注射生理鹽水,S-ly組:應(yīng)激組海馬內(nèi)注射LY294002。24小時(shí)后檢測小鼠行為和空間學(xué)習(xí)記憶能力各項(xiàng)指標(biāo)的變化,通過免疫組化實(shí)驗(yàn)檢測各組小鼠腦內(nèi)海馬和前額葉皮層BDNF、VEGF及PI3K表達(dá)的變化,以及通過HE染色,觀察海馬區(qū)和前額葉皮層神經(jīng)元的形態(tài)變化。研究結(jié)果:實(shí)驗(yàn)一:1.曠場實(shí)驗(yàn):與對(duì)照組相比,應(yīng)激組小鼠的爬行格數(shù)、修飾次數(shù)及直立次數(shù)均顯著減少,中央格停留時(shí)間顯著增加。2.懸尾實(shí)驗(yàn):與對(duì)照組相比,應(yīng)激組小鼠的第一次靜止不動(dòng)時(shí)間(潛伏期)顯著減少,后4分鐘累計(jì)不動(dòng)時(shí)間顯著增加。3.Morris水迷宮實(shí)驗(yàn):對(duì)照組和應(yīng)激組小鼠隨著訓(xùn)練次數(shù)的增加其逃避潛伏期和游泳總路程均縮短,對(duì)照組逃避潛伏期和游泳總路程均顯著低于應(yīng)激組;空間搜索實(shí)驗(yàn)中應(yīng)激組小鼠在目標(biāo)象限的停留時(shí)間顯著少于對(duì)照組。4.免疫組化實(shí)驗(yàn),與對(duì)照組相比,應(yīng)激組小鼠DG區(qū)、CA1區(qū)、CA3區(qū)及前額葉皮層的BDNF和VEGF表達(dá)均顯著降低,PI3K的表達(dá)在DG區(qū)和前額葉皮層顯著降低。表明,慢性應(yīng)激可引起小鼠海馬和前額葉皮層BDNF、VEGF和PI3K的表達(dá)降低。實(shí)驗(yàn)二:1.曠場實(shí)驗(yàn):相比于C-ns組,C-ly組小鼠的爬行格數(shù)、修飾次數(shù)及直立次數(shù)均顯著減少,S-ns組小鼠的爬行格數(shù)、修飾次數(shù)顯著減少;與S-ns組相比,S-ly組小鼠中央格停留時(shí)間、爬行格數(shù)、修飾次數(shù)均顯著減少。2.懸尾實(shí)驗(yàn):與C-ns組相比,C-ly組小鼠的第一次靜止不動(dòng)時(shí)間顯著減少,后4分鐘累計(jì)不動(dòng)時(shí)間顯著增加,S-ns組小鼠后4分鐘累計(jì)不動(dòng)時(shí)間顯著增加;與S-ns組相比,S-ly組小鼠后4分鐘累計(jì)不動(dòng)時(shí)間顯著增加。3.Morris水迷宮實(shí)驗(yàn):隨著訓(xùn)練次數(shù)的增加,C-ns組和S-ns組小鼠定位航行實(shí)驗(yàn)的逃避潛伏期和游泳總路程變化顯著,C-ly組和S-ly組沒有顯著變化。組間對(duì)比發(fā)現(xiàn),與C-ns組相比,C-ly組、S-ns組小鼠的逃避潛伏期和總路程均顯著增高;S-ly組小鼠的逃避潛伏期和游泳總路程均顯著高于S-ns組�？臻g搜索實(shí)驗(yàn)中,與C-ns組相比,C-ly組和S-ns組小鼠在目標(biāo)象限的停留時(shí)間顯著降低;S-ly組小鼠在目標(biāo)象限的停留時(shí)間顯著低于S-ns組。4.免疫組化實(shí)驗(yàn):與C-ns組相比,C-ly組小鼠海馬的DG區(qū)、CA1區(qū)和CA3區(qū)內(nèi)的BDNF、VEGF表達(dá)均顯著下降,DG區(qū)和前額葉皮層內(nèi)的PI3K表達(dá)顯著下降;與S-ns組相比,S-ly組小鼠海馬DG區(qū)、CA1區(qū)、CA3區(qū)和前額葉皮層內(nèi)的BDNF、PI3K表達(dá)均顯著下降,而VEGF在海馬的DG區(qū)、CA1區(qū)和CA3區(qū)表達(dá)顯著下降。表明,慢性應(yīng)激后雙側(cè)海馬注射PI3K-Akt信號(hào)通路阻斷劑LY294002后可導(dǎo)致BDNF、VEGF和PI3K的表達(dá)顯著降低。研究結(jié)論:1.慢性應(yīng)激可引起小鼠明顯的焦慮、抑郁樣行為,空間學(xué)習(xí)記憶能力減退;海馬內(nèi)注射LY294002后小鼠的行為和學(xué)習(xí)記憶功能損傷更嚴(yán)重。2.慢性應(yīng)激后小鼠海馬和前額葉皮層BDNF、VEGF和PI3K的表達(dá)降低;海馬內(nèi)注射LY294002后小鼠BDNF、VEGF和PI3K的表達(dá)降低更顯著。3.海馬和前腦皮層BDNF和VEGF表達(dá)的下調(diào)與慢性應(yīng)激所致小鼠的抑郁密切相關(guān),且可能通過PI3K/Akt信號(hào)通路參與抑郁癥的發(fā)生。
[Abstract]:Objective: to establish a model of chronic unpredictable stress depression in mice, to study the expression of VEGF and BDNF in the hippocampus (DG region, CA1 region, CA3 area) and prefrontal cortex in the brain of the depressive model mice and their relationship with the occurrence of depression, and to explore its role in the occurrence of depression and its related molecular mechanism. The research method: the 2 month old Kunming system was randomly selected. The mice in the control group were divided into the control group and the stress group. The mice in the control group were reared normally. The mice in the stress group adopted 7 different stress factors (water prohibition, fasting, high platform, oblique cage, foot shock, ice water swimming, all night lighting) to establish a chronic unpredictable stress and depression model. A test of open field experiment, tail suspension experiment, and Morris water maze experiment was used. The changes in the behavior of mice and the ability of spatial learning and memory were detected. The changes in the expression of BDNF, VEGF and PI3K in the hippocampus (DG area, CA1 area, CA3 area) and prefrontal cortex were detected by immunohistochemistry, and the morphological changes of neurons in the hippocampus and prefrontal cortex were observed by HE staining. Experiment two: the use of brain Stereotactic technique, the mice bilateral hippocampus was injected with saline and LY294002.C-ns group respectively: the control group was injected with saline (NS) in the hippocampus, group C-ly: the control group was injected with LY294002 in the hippocampus, S-ns group: the stress group was injected with saline in the hippocampus, group S-ly: the behavior and spatial learning and memory of the mice were detected after the hippocampus injection LY294002.24 hours in the stress group. Changes in various indexes, the changes in the expression of BDNF, VEGF and PI3K in the hippocampus and prefrontal cortex of the mice were detected by immunohistochemistry, and the morphological changes in the hippocampal and prefrontal cortex neurons were observed by HE staining. The results were as follows: Experiment 1: 1. open field experiment: the creeping number of mice in the stress group, compared with the control group, was compared with the control group. The number of modification and erect times were significantly reduced, and the residence time of the central lattice was significantly increased by the.2. suspension experiment. Compared with the control group, the first static time (incubation period) of the mice in the stress group decreased significantly, and the cumulative time of the latter 4 minutes increased significantly in the.3.Morris water maze test: the control group and the stress group increased with the increasing number of training times. The escape latency and the total swimming course were shortened, the escape latency and the total swimming course of the control group were significantly lower than those in the stress group; the time of the stress group in the target quadrant of the stress group was significantly less than the.4. immunization test in the control group. Compared with the control group, the mice in the stress group, the CA1 area, the CA3 region and the BDN of the prefrontal cortex were BDN. The expression of F and VEGF decreased significantly, and the expression of PI3K decreased significantly in the DG and prefrontal cortex. It showed that chronic stress could cause the expression of BDNF, VEGF and PI3K in the hippocampus and prefrontal cortex of mice. Experiment two: 1. open field experiment: the number of creeping, the number of modification and the erect times of the C-ly group were significantly decreased compared to the C-ns group, and the S-ns mice were significantly reduced. The number of crawling lattices and the number of modification decreased significantly. Compared with the S-ns group, the residence time of the central lattice, the number of creeping and the number of modifications in the S-ly group reduced the.2. suspension test significantly: compared with the C-ns group, the first time of static immobility in the C-ly group was significantly reduced, the cumulative time for the latter 4 minutes increased significantly, and the cumulative effect of the S-ns group was not moved after 4 minutes. Compared with the S-ns group, the cumulative time of 4 minutes after the S-ly group was significantly increased by the.3.Morris water maze experiment. As the number of training times increased, the escape latency and the total swimming distance of the C-ns and S-ns mice were significantly changed, and there was no significant change in the C-ly and S-ly groups. The escape latency and total route of mice in group C-ly and group S-ns were significantly higher than that in group S-ly. The escape latency and total swimming course of mice in group S-ly were significantly higher than that in group S-ns. Compared with the C-ns group, the stay time of the C-ly and S-ns mice in the target quadrant was significantly lower than that in the C-ns group; the time for the S-ly mice to stay in the target quadrant was significant. Compared with group.4., the expression of VEGF in DG area, CA1 area and CA3 area in group C-ly was significantly decreased, and PI3K expression in the DG area and prefrontal cortex decreased significantly compared with the group C-ns, and the expression in the hippocampus of the hippocampus, the region of the hippocampus, the frontal cortex and the cortex of the prefrontal cortex decreased significantly compared with those in the S-ns group. The expression of VEGF in the hippocampal DG region, CA1 area and CA3 region decreased significantly. The results showed that the expression of BDNF, VEGF and PI3K decreased significantly after the injection of PI3K-Akt signaling blocking agent LY294002 after chronic stress. The conclusion: 1. chronic stress can cause obvious anxiety, depressive behavior, spatial learning and memory impairment in mice; hippocampus After intramuscular injection of LY294002, the behavior and learning and memory function of mice were more severe. The expression of BDNF, VEGF and PI3K in the hippocampus and prefrontal cortex of mice decreased after.2. chronic stress, and the expression of BDNF, VEGF and PI3K decreased more significantly in the hippocampus and the lower.3. hippocampus and the downregulation of BDNF and VEGF expressions in the hippocampus and the prefrontal cortex and the decrease of chronic stress and chronic stress. Depression is closely related to mice, and may participate in the occurrence of depression through the PI3K/Akt signaling pathway.
【學(xué)位授予單位】：曲阜師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R749.4;R-332
本文編號(hào)：2153110
[Abstract]:Objective: to establish a model of chronic unpredictable stress depression in mice, to study the expression of VEGF and BDNF in the hippocampus (DG region, CA1 region, CA3 area) and prefrontal cortex in the brain of the depressive model mice and their relationship with the occurrence of depression, and to explore its role in the occurrence of depression and its related molecular mechanism. The research method: the 2 month old Kunming system was randomly selected. The mice in the control group were divided into the control group and the stress group. The mice in the control group were reared normally. The mice in the stress group adopted 7 different stress factors (water prohibition, fasting, high platform, oblique cage, foot shock, ice water swimming, all night lighting) to establish a chronic unpredictable stress and depression model. A test of open field experiment, tail suspension experiment, and Morris water maze experiment was used. The changes in the behavior of mice and the ability of spatial learning and memory were detected. The changes in the expression of BDNF, VEGF and PI3K in the hippocampus (DG area, CA1 area, CA3 area) and prefrontal cortex were detected by immunohistochemistry, and the morphological changes of neurons in the hippocampus and prefrontal cortex were observed by HE staining. Experiment two: the use of brain Stereotactic technique, the mice bilateral hippocampus was injected with saline and LY294002.C-ns group respectively: the control group was injected with saline (NS) in the hippocampus, group C-ly: the control group was injected with LY294002 in the hippocampus, S-ns group: the stress group was injected with saline in the hippocampus, group S-ly: the behavior and spatial learning and memory of the mice were detected after the hippocampus injection LY294002.24 hours in the stress group. Changes in various indexes, the changes in the expression of BDNF, VEGF and PI3K in the hippocampus and prefrontal cortex of the mice were detected by immunohistochemistry, and the morphological changes in the hippocampal and prefrontal cortex neurons were observed by HE staining. The results were as follows: Experiment 1: 1. open field experiment: the creeping number of mice in the stress group, compared with the control group, was compared with the control group. The number of modification and erect times were significantly reduced, and the residence time of the central lattice was significantly increased by the.2. suspension experiment. Compared with the control group, the first static time (incubation period) of the mice in the stress group decreased significantly, and the cumulative time of the latter 4 minutes increased significantly in the.3.Morris water maze test: the control group and the stress group increased with the increasing number of training times. The escape latency and the total swimming course were shortened, the escape latency and the total swimming course of the control group were significantly lower than those in the stress group; the time of the stress group in the target quadrant of the stress group was significantly less than the.4. immunization test in the control group. Compared with the control group, the mice in the stress group, the CA1 area, the CA3 region and the BDN of the prefrontal cortex were BDN. The expression of F and VEGF decreased significantly, and the expression of PI3K decreased significantly in the DG and prefrontal cortex. It showed that chronic stress could cause the expression of BDNF, VEGF and PI3K in the hippocampus and prefrontal cortex of mice. Experiment two: 1. open field experiment: the number of creeping, the number of modification and the erect times of the C-ly group were significantly decreased compared to the C-ns group, and the S-ns mice were significantly reduced. The number of crawling lattices and the number of modification decreased significantly. Compared with the S-ns group, the residence time of the central lattice, the number of creeping and the number of modifications in the S-ly group reduced the.2. suspension test significantly: compared with the C-ns group, the first time of static immobility in the C-ly group was significantly reduced, the cumulative time for the latter 4 minutes increased significantly, and the cumulative effect of the S-ns group was not moved after 4 minutes. Compared with the S-ns group, the cumulative time of 4 minutes after the S-ly group was significantly increased by the.3.Morris water maze experiment. As the number of training times increased, the escape latency and the total swimming distance of the C-ns and S-ns mice were significantly changed, and there was no significant change in the C-ly and S-ly groups. The escape latency and total route of mice in group C-ly and group S-ns were significantly higher than that in group S-ly. The escape latency and total swimming course of mice in group S-ly were significantly higher than that in group S-ns. Compared with the C-ns group, the stay time of the C-ly and S-ns mice in the target quadrant was significantly lower than that in the C-ns group; the time for the S-ly mice to stay in the target quadrant was significant. Compared with group.4., the expression of VEGF in DG area, CA1 area and CA3 area in group C-ly was significantly decreased, and PI3K expression in the DG area and prefrontal cortex decreased significantly compared with the group C-ns, and the expression in the hippocampus of the hippocampus, the region of the hippocampus, the frontal cortex and the cortex of the prefrontal cortex decreased significantly compared with those in the S-ns group. The expression of VEGF in the hippocampal DG region, CA1 area and CA3 region decreased significantly. The results showed that the expression of BDNF, VEGF and PI3K decreased significantly after the injection of PI3K-Akt signaling blocking agent LY294002 after chronic stress. The conclusion: 1. chronic stress can cause obvious anxiety, depressive behavior, spatial learning and memory impairment in mice; hippocampus After intramuscular injection of LY294002, the behavior and learning and memory function of mice were more severe. The expression of BDNF, VEGF and PI3K in the hippocampus and prefrontal cortex of mice decreased after.2. chronic stress, and the expression of BDNF, VEGF and PI3K decreased more significantly in the hippocampus and the lower.3. hippocampus and the downregulation of BDNF and VEGF expressions in the hippocampus and the prefrontal cortex and the decrease of chronic stress and chronic stress. Depression is closely related to mice, and may participate in the occurrence of depression through the PI3K/Akt signaling pathway.
【學(xué)位授予單位】：曲阜師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R749.4;R-332
【參考文獻(xiàn)】
相關(guān)期刊論文 前10條
1 王睿;黃樹明;;抑郁癥發(fā)病機(jī)制研究進(jìn)展[J];醫(yī)學(xué)研究生學(xué)報(bào);2014年12期
2 黃致嘉;王立偉;;血管內(nèi)皮生長因子與抑郁癥的神經(jīng)營養(yǎng)假說[J];國際精神病學(xué)雜志;2014年01期
3 劉春林;阮克鋒;高君偉;吳飛;張繼全;;抑郁癥的多機(jī)制發(fā)病[J];生理科學(xué)進(jìn)展;2013年04期
4 樓劍書;楊曉春;方杰;潘建立;郭建友;;免疫激活對(duì)抑郁癥谷氨酸和五羥色胺系統(tǒng)的調(diào)節(jié)[J];中國藥理學(xué)通報(bào);2009年12期
5 薛瑞;張有志;鄒莉波;;快速起效抗抑郁藥物的研究進(jìn)展[J];中國藥理學(xué)通報(bào);2008年12期
6 季寧東;李娟好;李明亞;郭麗冰;管華基;;石菖蒲提取液的抗抑郁作用及柴胡皂苷對(duì)其作用的影響[J];南京醫(yī)科大學(xué)學(xué)報(bào)(自然科學(xué)版);2006年12期
7 孫春艷;胡豫;吳濤;王雅丹;王華芳;何文娟;;腦源性神經(jīng)營養(yǎng)因子對(duì)血管新生的作用[J];中華病理學(xué)雜志;2006年04期
8 張峰,曹仲偉,張學(xué)杰,李法曾;柴胡對(duì)大鼠慢性應(yīng)激抑郁模型腦單胺類神經(jīng)遞質(zhì)及其代謝物含量的影響[J];山東中醫(yī)藥大學(xué)學(xué)報(bào);2005年03期
9 儲(chǔ)智勇,湯文,卞俊,姜鐵夫;貫葉金絲桃提取物對(duì)行為絕望鼠抑郁模型的抗抑郁作用[J];解放軍藥學(xué)學(xué)報(bào);2003年06期
10 邢偉,孫黎光,時(shí)利德,劉素嬡,蔡葵,滕國璽,蔡原,郭納新,劉秋芳,張穎花,奚奇,萬伯健;染鉛鼠腦海馬PKC活性與LTP的相關(guān)性研究[J];衛(wèi)生毒理學(xué)雜志;1996年03期
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