【摘要】：目的：通過(guò)建立急性CO中毒遲發(fā)型腦�。―EACMP）大鼠模型，觀察HIF-1α及EPO在相應(yīng)動(dòng)物模型中的表達(dá)變化，探討HIF-1α及EPO在DEACMP發(fā)病機(jī)制中的作用。方法：78只健康雄性成年SD大鼠（SPF級(jí)），體重在240～280g之間，隨機(jī)分為三組：空白對(duì)照組（BC組）18只、空氣對(duì)照組（AC組）30只、急性一氧化碳中毒遲發(fā)性腦病組（CO組）30只。其中CO組采用腹腔注射法建立DEACMP大鼠模型，AC組采用腹腔注射法注射等量空氣，BC組不進(jìn)行任何手術(shù)操作。按中毒后1d、3d、7d、14d、21d、28d六個(gè)時(shí)間點(diǎn)將三個(gè)組各分為6個(gè)亞組，其中CO組和AC組每個(gè)亞組5只大鼠，BC組每個(gè)亞組3只大鼠。觀察大鼠急性CO中毒表現(xiàn)和大鼠動(dòng)脈血COHb濃度及中毒后期運(yùn)用Morris水迷宮實(shí)驗(yàn)檢測(cè)大鼠認(rèn)知行為學(xué)改變。各組大鼠的腦組織切片均應(yīng)用HE染色觀察大鼠海馬區(qū)病理變化，用TUNEL染色檢測(cè)海馬區(qū)錐體細(xì)胞凋亡情況，應(yīng)用免疫組化染色檢測(cè)海馬區(qū)HIF-1α、EPO蛋白的表達(dá)。結(jié)果：1、Morris水迷宮實(shí)驗(yàn)結(jié)果：CO組大鼠在染毒后出現(xiàn)平均逃避潛伏期延長(zhǎng)，且在染毒后14d、21d及28d時(shí)，CO組較BC組和AC組比較差異均有顯著統(tǒng)計(jì)學(xué)意義（P0.01)。染毒后CO組大鼠在第Ⅳ象限運(yùn)動(dòng)時(shí)間有縮短趨勢(shì)，且染毒14d及染毒28d時(shí)，CO組較BC組及AC組比較差異有統(tǒng)計(jì)學(xué)意義（P0.05)。染毒后CO組大鼠穿越平臺(tái)次數(shù)有降低趨勢(shì)，染毒28d時(shí)，CO組穿越平臺(tái)次數(shù)較其余兩組低，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。2、HE染色：CO組大鼠在急性CO中毒后出現(xiàn)海馬區(qū)錐體細(xì)胞數(shù)目減少、細(xì)胞排列松散、細(xì)胞水腫、核固縮、核碎裂等病理學(xué)改變。3、TUNEL法檢測(cè)：CO組大鼠海馬區(qū)錐體細(xì)胞于染毒3d時(shí)凋亡開始增多，，7d時(shí)達(dá)到高峰，28d時(shí)仍有少量細(xì)胞凋亡，各時(shí)間點(diǎn)凋亡指數(shù)與BC組及AC組比較，差異具有顯著性（P0.01)。4、免疫組化染色結(jié)果：1）HIF-1α蛋白測(cè)定：HIF-1α在BC組及AC組中表達(dá)較少，且各時(shí)間點(diǎn)間差異不具有顯著性，CO組在染毒1d時(shí)出現(xiàn)HIF-1α的表達(dá)升高，3d達(dá)到峰值，28d時(shí)仍有表達(dá)，表達(dá)量明顯高于BC組及AC組，差異具有顯著性（P0.01）；2）EPO蛋白測(cè)定：在BC組及AC組中有少量表達(dá)，且各時(shí)間點(diǎn)間差異不具有顯著性，CO組在染毒1d時(shí)明顯升高，3d為高峰，7d時(shí)下降，染毒1d、3d、7d、14d時(shí)CO組較BC組與AC組差異具有顯著性（P0.01）。結(jié)論：1.腹腔注射CO法制作DEACMP大鼠模型，操作簡(jiǎn)單，是研究DEACMP較為理想的動(dòng)物模型；2.Morris水迷宮實(shí)驗(yàn)是初步判斷大鼠DEACMP較為可靠的依據(jù)；3.HIF-1α可能通過(guò)誘導(dǎo)急性CO中毒后海馬區(qū)神經(jīng)元的遲發(fā)性凋亡而參與DEACMP的發(fā)�。�4.EPO在急性CO中毒后中后期神經(jīng)保護(hù)作用減弱，可能是導(dǎo)致DEACMP發(fā)病原因之一。
[Abstract]:Objective: to investigate the role of HIF-1 偽 and EPO in the pathogenesis of DEACMP by establishing a rat model of delayed encephalopathy (DEACMP) caused by acute CO poisoning, and observing the expression of HIF-1 偽 and EPO in corresponding animal models. Methods Seventy-eight healthy male adult SD rats (SPF grade), weighing between 240 and 280 g, were randomly divided into three groups: blank control group (BC group, n = 18), air control group (AC group, n = 30) and acute carbon monoxide poisoning delayed encephalopathy group (CO group, n = 30). In the CO group, the DEACMP rat model was established by intraperitoneal injection. The AC group was injected the same amount of air by intraperitoneal injection without any operation. The three groups were divided into 6 subgroups according to the six time points 1 day, 3 days, 7 days, 14 days and 21 days, 28 days after poisoning. Five rats in each subgroup of CO group and AC group were divided into 3 rats in each subgroup of BC group. The manifestations of acute CO poisoning and the concentration of COHb in arterial blood of rats were observed. Morris water maze test was used to detect the changes of cognitive behavior in rats. The pathological changes of hippocampus were observed by HE staining, apoptosis of pyramidal cells in hippocampus were detected by Tunel staining, and HIF-1 偽 EPO protein expression in hippocampus was detected by immunohistochemical staining. Results in the water maze experiment, the escape latency of the rats in the control group was longer than that in the BC group and AC group (P0.01), and there was a significant difference between the CO group and the BC group on the 21st day and the 28th day after exposure (P0.01). The time of exercise in the fourth quadrant was shortened in CO group after exposure, and there was significant difference between CO group and BC group and AC group in 14 days and 28 days (P0.05). The number of crossing the platform in CO group was lower than that in the other two groups after 28 days of exposure (P0.05). The number of pyramidal cells in hippocampus decreased after acute CO poisoning. Apoptosis of pyramidal cells in hippocampal area of rats in the 30% CO group was detected by Tunel method. Apoptosis began to increase on the 3rd day and reached the peak on the 7th day and reached the peak at the end of 28 days, and there was still a small amount of apoptosis in the hippocampal pyramidal cells of the rats in the control group. Compared with BC group and AC group, the apoptotic index at each time point had significant difference (P0.01) .4.The result of immunohistochemical staining was: 1) the expression of HIF-1 偽 protein in BC group and AC group was lower than that in BC group and AC group, and the expression of HIF-1 偽 protein was lower in BC group and AC group than in BC group and AC group. The expression of HIF-1 偽 in CO group was significantly higher than that in BC group and AC group (P0.01), and the expression of HIF-1 偽 in CO group was significantly higher than that in BC group and AC group (P0.01), and the expression of HIF-1 偽 in CO group was significantly higher than that in BC group and AC group (P0.01), and the expression of HIF-1 偽 in CO group was significantly higher than that in BC group (P0.01). 2) determination of EPO protein: there was a little expression of EPO protein in BC group and AC group, and there was no significant difference between different time points. The level of EPO protein in CO group was significantly higher than that in BC group and AC group at 1 day after exposure to CO for 1 day, and decreased at the peak of 3 days and 7 days after exposure to CO (P0.01), and there was a significant difference between CO group and AC group at 14 days after exposure (P0.01). Conclusion 1. The method of intraperitoneal injection of CO to make DEACMP rat model is simple, which is an ideal animal model for DEACMP. 2. Morris water maze test is a reliable basis for preliminary judgement of DEACMP in rats. 3. HIF-1 偽 may be involved in the pathogenesis of DEACMP by inducing delayed apoptosis of hippocampal neurons after acute CO poisoning. 4. The neuroprotective effect of EPO in the middle and late stages of acute CO poisoning may be one of the causes of DEACMP.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R747.9

【共引文獻(xiàn)】

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3 李丹;段宣初;;促紅細(xì)胞生成素對(duì)視神經(jīng)視網(wǎng)膜的保護(hù)作用[J];國(guó)際眼科雜志;2006年04期

4 曹娟;賈天明;李文霞;代紅;;電刺激對(duì)缺氧缺血性腦損傷新生大鼠腦組織血管內(nèi)皮細(xì)胞生長(zhǎng)因子及其受體表達(dá)的影響[J];實(shí)用兒科臨床雜志;2010年01期

5 郭麗君;袁慧欣;闞衛(wèi)軍;喬振華;薛福平;;重組人紅細(xì)胞生成素對(duì)大鼠腦缺血再灌注損傷后XIAP及Smac蛋白表達(dá)的影響[J];中西醫(yī)結(jié)合心腦血管病雜志;2011年01期

6 刁孟元;張斌;黃鳳樓;;血管內(nèi)皮生長(zhǎng)因子與海水淹溺型急性肺損傷的關(guān)系[J];中國(guó)療養(yǎng)醫(yī)學(xué);2013年03期

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2 熊思齊;促紅細(xì)胞生成素在視網(wǎng)膜新生血管形成中調(diào)控作用的研究[D];中南大學(xué);2009年

3 王輝;重組人促紅細(xì)胞生成素對(duì)大鼠視神經(jīng)保護(hù)作用的實(shí)驗(yàn)研究[D];中國(guó)醫(yī)科大學(xué);2008年

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1 陳炳達(dá);急性一氧化碳中毒后遲發(fā)性腦病大鼠海馬區(qū)HIF-1α及VEGF蛋白動(dòng)態(tài)表達(dá)的實(shí)驗(yàn)研究[D];瀘州醫(yī)學(xué)院;2014年

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