發(fā)布時(shí)間：2018-03-03 21:38

  本文選題：豐富環(huán)境　切入點(diǎn)：缺氧缺血性腦損傷　出處：《青島大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的本研究擬通過(guò)制作7日齡(相當(dāng)于人類新生兒期)缺氧缺血性腦損傷(hypoxic-ischemic brain damage,HIBD)新生大鼠模型,早期給予不同強(qiáng)度豐富環(huán)境(environmental enrichment,EE)干預(yù),動(dòng)態(tài)觀察在不同干預(yù)強(qiáng)度下HIBD大鼠學(xué)習(xí)記憶能力、海馬區(qū)突觸可塑性相關(guān)分子(腦源性神經(jīng)營(yíng)養(yǎng)因子、突觸素蛋白)的表達(dá)變化,以探討HIBD大鼠早期豐富環(huán)境干預(yù)的有效性及機(jī)制,尤其是不同干預(yù)強(qiáng)度對(duì)HIBD大鼠學(xué)習(xí)記憶功能、海馬突觸可塑性的影響及其分子機(jī)制,從而為臨床HIBD患兒豐富環(huán)境的構(gòu)建及康復(fù)方案的制定提供可能的理論依據(jù),指導(dǎo)臨床實(shí)踐。方法將40只7日齡Wistar雄性大鼠隨機(jī)分為以下四組:6小時(shí)豐富環(huán)境干預(yù)組(6h EE)、12小時(shí)豐富環(huán)境干預(yù)組(12h EE)、模型組(Model)和假手術(shù)組(Sham),每組均10只。豐富環(huán)境干預(yù)組和模型組均運(yùn)用Rice法(1981)制備大鼠缺氧缺血腦損傷模型。豐富環(huán)境干預(yù)組分別進(jìn)行14天的6h和12h豐富環(huán)境干預(yù)。干預(yù)結(jié)束后運(yùn)用Morris水迷宮行為學(xué)實(shí)驗(yàn)測(cè)查大鼠的學(xué)習(xí)記憶能力。運(yùn)用免疫印跡(Western blot)法檢測(cè)大鼠海馬區(qū)腦源性神經(jīng)營(yíng)養(yǎng)因子(BDNF)、突觸素蛋白的表達(dá)。結(jié)果1、水迷宮實(shí)驗(yàn)的定位航行實(shí)驗(yàn)中,4組大鼠訓(xùn)練期間的逃避潛伏期隨著訓(xùn)練天數(shù)的增加逐漸降低,但4組大鼠之間的逃避潛伏期比較差異無(wú)統(tǒng)計(jì)學(xué)意義(F=0.2369,P0.05)。2、水迷宮實(shí)驗(yàn)中,第5天空間探索測(cè)試時(shí),模型組在目標(biāo)象限尋找平臺(tái)的時(shí)間與在其他象限比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);6h EE組、12h EE組、假手術(shù)組在目標(biāo)象限尋找平臺(tái)的時(shí)間顯著長(zhǎng)于其他象限,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。3、水迷宮實(shí)驗(yàn)中,第7天空間探索測(cè)試時(shí),4組大鼠在目標(biāo)象限尋找平臺(tái)的時(shí)間都長(zhǎng)于其他象限,說(shuō)明4組大鼠均能記住平臺(tái)所在象限位置。6h豐富環(huán)境干預(yù)組(((?)±s)6h EE=38.132±2.876,P0.001)、12h豐富環(huán)境干預(yù)組(((?)±s)12h EE=45.526±2.648,P0.001)和模型組(((?)±s)_(Model)=30.702±3.819,P0.001)的新生大鼠在目標(biāo)象限尋找平臺(tái)的時(shí)間均短于假手術(shù)組(54.400±3.152),并且海馬區(qū)BDNF(((?)±s)6h EE=0.529±4、0.038,P0.001;((?)±s)12h EE=0.660±0.034,P0.01;((?)±s)_(Model)=0.225±0.015,P0.001)和突觸素蛋白(((?)±s)6h EE=0.889±0.027,P0.001;((?)±s)12h EE=1.114±0.037,P0.05;((?)±s)_(Model)=0.672±0.057,P0.001)的表達(dá)相比于假手術(shù)組(((?)±s)BDNF=0.803±0.026;5、((?)±s)突觸素=1.347±0.092)也有所下降。6、與模型組相比,6h豐富環(huán)境干預(yù)組和12h豐富環(huán)境干預(yù)組的新生大鼠在目標(biāo)象限尋找平臺(tái)的時(shí)間更長(zhǎng)(P6h EE0.01;P12h EE0.001),并且海馬區(qū)BDNF(P6h EE0.001;P12h EE0.001)和突觸素蛋白(P6h EE0.05;P12h EE0.001)的表達(dá)顯著上升。7、更值得提及的是12h豐富環(huán)境干預(yù)組在目標(biāo)象限尋找平臺(tái)的時(shí)間(P0.01)以及海馬區(qū)BDNF(P0.01)與突觸素蛋白(P0.05)的表達(dá)情況都優(yōu)于6h豐富環(huán)境干預(yù)組。結(jié)論豐富環(huán)境能夠改善缺氧缺血性腦損傷新生大鼠的學(xué)習(xí)記憶能力,保護(hù)神經(jīng)元細(xì)胞,其機(jī)制可能與上調(diào)海馬區(qū)BDNF和突觸素蛋白表達(dá)有關(guān)。
[Abstract]:Objective to establish a neonatal rat model of hypoxic-ischemic brain damage (HIBD) with hypoxic-ischemic brain damage (HIBD) at 7 days of age (equivalent to human newborn), and to give early intervention with environmental hazards of different intensities. The learning and memory ability of HIBD rats and the expression of synaptic plasticity related molecules (brain-derived neurotrophic factor, synaptophysin protein) in hippocampus were observed dynamically under different intervention intensity. To explore the effectiveness and mechanism of early rich environment intervention in HIBD rats, especially the effects of different intervention intensities on learning and memory function, hippocampal synaptic plasticity and its molecular mechanism in HIBD rats. Thus, it can provide a theoretical basis for the construction of rich environment and the establishment of rehabilitation program for children with HIBD. Methods 40 7-day-old Wistar male rats were randomly divided into the following four groups: 6 hour enriched environment intervention group (6 hours) and 12 hour enriched environment intervention group (12 h EEN, model group) and sham operation group (10 rats in each group). The model of hypoxic-ischemic brain injury in rats was established by Rice method in both the intervention group and the model group. The rich environment intervention group was treated with 14 days of rich environment intervention for 6 hours and 12 hours, respectively. After the intervention, Morris water maze test was used to test the behavior of rats. The expression of brain-derived neurotrophic factor (BDNF) and synaptophysin (synaptophysin) in the hippocampus of rats was detected by Western blot method. The escape latency decreased with the increase of training days. However, there was no significant difference in the escape latency among the four groups. In the water maze experiment, the time of finding the platform in the target quadrant in the model group was compared with that in the other quadrants during the space exploration test on the 5th day in the water maze experiment. There was no significant difference in the time of searching platform in the target quadrant between the sham operation group and the other quadrants. The difference was statistically significant (P 0.05). 3. In the water maze experiment, there was no significant difference between the two groups. On the 7th day of space exploration test, the time of searching platform in the target quadrant was longer in the 4 groups than in other quadrants, indicating that the four groups could remember the location of the platform in the quadrant. EE=38.132 鹵2.876% P0.001 + 12h enriched environmental intervention group (EE=38.132 鹵2.876 P0.001) for 12 hours. ) 12 h EE=45.526 鹵2.648 (P0.001) and model group (P 0.001). The time of finding platform in the target quadrant of the neonatal rats was shorter than that of the sham-operated group (54.400 鹵3.152) and the hippocampal area of BDNF? EE=0.529 鹵4 0. 038? P0. 001? ) 12 h EE=0.660 鹵0.034 P0.01? ) and synaptophysin? 6 h EE=0.889 鹵0.027 P0.001? ) 12 h EE=1.114 鹵0.037 P0.05? Compared with sham operation group (P 0.001), the expression of 0.672 鹵0.057 (P 0.001) was significantly higher than that of sham operation group (P < 0.05). ) BDNF 0. 803 鹵0. 026? ) 鹵s) synaptophysin 1. 347 鹵0. 092) was also decreased. Compared with the model group, the neonatal rats in the 6 h enriched environment intervention group and the 12 hour enriched environment intervention group had longer time searching for platform in the target quadrant, and the hippocampal area BDNF(P6h EE0. 001 P12 h EE0. 001) and synaptophysin protein. The expression of BDNFP0.01) and synaptophysin protein (P0.05) in hippocampus of 12h enriched environment intervention group were significantly higher than that of 6h rich environment intervention group (P0.01), and the expression of synaptophysin protein P0.05) was better than that of 6-hour enriched environment intervention group. Rich environment can improve the learning and memory ability of neonatal rats with hypoxic-ischemic brain injury. The protective mechanism of neuronal cells may be related to the up-regulation of BDNF and synaptophysin expression in hippocampal area.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R742

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