本文選題：缺血預(yù)適應(yīng)　切入點(diǎn)：腦缺血再灌注　出處：《河北聯(lián)合大學(xué)》2014年碩士論文

【摘要】：目的觀察缺血預(yù)適應(yīng)對(duì)局灶性腦缺血再灌注大鼠缺血灶周圍皮質(zhì)區(qū)及缺血側(cè)海馬CA1區(qū)低氧誘導(dǎo)因子-1α（HIF-1α）及存活素（Survivin）表達(dá)的影響，，探討缺血預(yù)適應(yīng)的腦保護(hù)機(jī)制。 方法130只雄性SD大鼠，隨機(jī)分為假手術(shù)組（SO組，n=10）、腦缺血組（MCAO組，n=60）、腦缺血預(yù)適應(yīng)組（BIP組，n=60），后兩組再分為缺血2h再灌注2h、6h、12h、24h、48h、72h6個(gè)觀察時(shí)間點(diǎn)（每個(gè)時(shí)間點(diǎn)各10只，其中5只用于免疫組化染色，另外5只用于Western blot檢測(cè)）。采用大腦中動(dòng)脈兩次線栓法制備大鼠腦缺血預(yù)適應(yīng)模型。各組作如下處理：BIP組：大腦中動(dòng)脈缺血10min為預(yù)適應(yīng)條件，24h后行大腦中動(dòng)脈缺血2h；MCAO組：以假手術(shù)代替預(yù)適應(yīng)，余步驟同BIP組；SO組：以兩次假手術(shù)代替預(yù)適應(yīng)及再缺血。免疫組織化學(xué)染色及Western blot法檢測(cè)缺血灶周圍皮質(zhì)區(qū)及缺血側(cè)海馬CA1區(qū)HIF-1α、Survivin的表達(dá)情況。 結(jié)果1SO組皮質(zhì)區(qū)HIF-1α陽性細(xì)胞表達(dá)水平低（4.77±1.11），蛋白表達(dá)量低（0.45±0.03）。MCAO組缺血灶周圍皮質(zhì)區(qū)HIF-1α陽性細(xì)胞、缺血側(cè)皮質(zhì)區(qū)HIF-1α蛋白量表達(dá)均于腦缺血再灌注2h升高，24h達(dá)各時(shí)間點(diǎn)最高水平，隨后表達(dá)逐漸降低，72h時(shí)仍明顯高于SO組。BIP組缺血灶周圍皮質(zhì)區(qū)HIF-1α陽性細(xì)胞及缺血側(cè)皮質(zhì)區(qū)HIF-1α蛋白量表達(dá)趨勢(shì)與MCAO組一致，相同時(shí)間點(diǎn)HIF-1α陽性細(xì)胞數(shù)量及蛋白表達(dá)量均高于MCAO組，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。MCAO組、BIP組HIF-1α陽性細(xì)胞數(shù)量及蛋白表達(dá)量均高于SO組，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。2SO組海馬CA1區(qū)HIF-1α陽性細(xì)胞的表達(dá)較低（4.80±1.30），蛋白表達(dá)量較低（0.35±0.03）。MCAO組缺血側(cè)海馬CA1區(qū)HIF-1α陽性細(xì)胞及蛋白量表達(dá)于腦缺血再灌注后2h升高，24h在各時(shí)間點(diǎn)表達(dá)水平最高，而后逐漸下降，72h時(shí)表達(dá)接近6h水平。BIP組缺血側(cè)海馬CA1區(qū)HIF-1α陽性細(xì)胞及蛋白量表達(dá)規(guī)律與MCAO組一致，除再灌注2h、6h外，相同時(shí)間點(diǎn)BIP組HIF-1α陽性細(xì)胞的數(shù)量高于MCAO組，差異有統(tǒng)計(jì)學(xué)意義（P0.05）；BIP組各時(shí)間點(diǎn)HIF-1α蛋白表達(dá)量均高于MCAO組，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。MCAO組、BIP組HIF-1α陽性細(xì)胞數(shù)量及蛋白表達(dá)量均高于SO組，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。3SO組皮質(zhì)區(qū)可見少量Survivin陽性細(xì)胞表達(dá)（4.96±0.63），少量Survivin蛋白表達(dá)（0.59±0.03）。MCAO組于腦缺血再灌注2h缺血灶周圍皮質(zhì)區(qū)Survivin陽性細(xì)胞、缺血側(cè)皮質(zhì)區(qū)Survivin蛋白量表達(dá)增加，48h達(dá)各時(shí)間點(diǎn)最高水平，72h時(shí)表達(dá)接近24h水平。BIP組缺血灶周圍皮質(zhì)區(qū)Survivin陽性細(xì)胞、缺血側(cè)皮質(zhì)區(qū)Survivin蛋白量的表達(dá)趨勢(shì)與MCAO組具備相同的時(shí)間規(guī)律性，除再灌注2h外，同一時(shí)間點(diǎn)BIP組Survivin陽性細(xì)胞表達(dá)數(shù)量高于MCAO組，差異有統(tǒng)計(jì)學(xué)意義（P0.05）；BIP組各時(shí)間點(diǎn)Survivin蛋白表達(dá)量較MCAO組高，兩組比較差異有統(tǒng)計(jì)學(xué)意義（P0.05）。MCAO組與BIP組Survivin陽性細(xì)胞的表達(dá)數(shù)量及蛋白表達(dá)量均高于SO組，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。4SO組海馬CA1區(qū)Survivin陽性細(xì)胞表達(dá)量少（3.18±0.46），蛋白表達(dá)量少（0.39±0.03）。MCAO組于腦缺血再灌注2h缺血側(cè)海馬CA1區(qū)Survivin陽性細(xì)胞及蛋白量表達(dá)增加，24h在各時(shí)間點(diǎn)水平為最高，隨后表達(dá)逐漸下降，72h時(shí)表達(dá)接近6h水平。BIP組缺血側(cè)海馬CA1區(qū)Survivin陽性細(xì)胞及蛋白量的表達(dá)趨勢(shì)與MCAO組時(shí)間性一致，且各時(shí)間點(diǎn)Survivin陽性細(xì)胞的表達(dá)數(shù)量及蛋白表達(dá)量均高于MCAO組，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。以上兩組Survivin陽性細(xì)胞及蛋白的表達(dá)水平均高于SO組，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論1缺血預(yù)適應(yīng)可以促進(jìn)局灶性腦缺血再灌注大鼠缺血灶周圍皮質(zhì)區(qū)及缺血側(cè)海馬CA1區(qū)HIF-1α、Survivin的表達(dá)。2缺血預(yù)適應(yīng)可能通過HIF-1α、Survivin的表達(dá)上調(diào)參與腦缺血耐受的發(fā)生。
[Abstract]:Objective To investigate the effects of ischemic preconditioning on the expression of hypoxia - inducible factor - 1偽 ( HIF - 1偽 ) and survivin ( Survivin ) in the peripheral cortex and hippocampus CA1 region of ischemia - reperfusion rats and to explore the mechanism of ischemic preconditioning .

Methods 130 male SD rats were randomly divided into sham operation group ( SO group , n = 10 ) , cerebral ischemia group ( middle cerebral ischemia group , n = 60 ) and cerebral ischemia preconditioning group ( n = 60 ) . Two groups were divided into two groups : ischemia 2h reperfusion 2h , 6h , 12h , 24h , 48h , 72h6 observation time points ( 10 at each time point , 5 only for immunohistochemical staining and 5 for Western blot detection ) . The model of cerebral ischemia preconditioning was prepared by using two - thread method of middle cerebral artery occlusion . The following treatments were used : the cerebral artery ischemia was 10 min as the preconditioning condition , and the middle cerebral artery was ischemia for 2h after 24h ;
In the middle group , the sham operation was used to replace the pre - adaptation , and the remaining steps were the same as that of the bip group .
SO group : The expression of HIF - 1偽 and Survivin was detected by two false surgeries instead of preconditioning and ischemia . Immunohistochemical staining and Western blot were used to detect the expression of HIF - 1偽 and Survivin in the cortex and hippocampus CA1 region .

Results The expression of HIF - 1偽 positive cells and the expression of HIF - 1偽 in the cortex were higher than those in the middle cerebral ischemia - reperfusion group ( P0.05 ) .
There was a significant difference in the expression of Survivin positive cells ( 4.96 鹵 0.63 ) and Survivin protein expression ( P 0.05 ) . The expression of Survivin positive cells in the cortical area around the ischemic foci was significantly higher than that in the middle cerebral ischemia group ( P 0.05 ) .
The expression of survivin - positive cells and protein in the CA1 region of the hippocampus CA1 region were significantly higher than those in the middle cerebral ischemia - reperfusion 2h group ( P0.05 ) .

Conclusion 1 Ischemia preconditioning can promote the expression of HIF - 1偽 and Survivin in the peripheral cortex and hippocampus CA1 region of focal cerebral ischemia - reperfusion rats . The expression of HIF - 1偽 and Survivin may be increased by the expression of HIF - 1偽 and Survivin in the ischemic preconditioning .

【學(xué)位授予單位】：河北聯(lián)合大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R743.3

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