【摘要】：目的： 研究亞低溫對(duì)大鼠全腦缺血再灌注性腦損傷的保護(hù)作用，并初步探討其機(jī)制。 方法： 選取成年雄性Wistar大鼠150只，實(shí)驗(yàn)大鼠隨機(jī)分為3組，每組50只，即分為假手術(shù)組（sham組）、10min缺血/再灌注組（10min I/R組）、亞低溫處理組。本實(shí)驗(yàn)采用大鼠全腦缺血再灌注模型，缺血時(shí)間為10min，再灌注時(shí)間為4h，24h，48h，72h，1周。通過(guò)甲醛固定腦組織并切片，通過(guò)HE染色運(yùn)用高倍顯微鏡觀察全腦缺血再灌注24h后的大鼠海馬CA1區(qū)神經(jīng)元細(xì)胞，比較各組間大鼠海馬CA1區(qū)神經(jīng)元存活數(shù)量；達(dá)到再灌注時(shí)間后，通過(guò)改良的NSS神經(jīng)功能評(píng)分標(biāo)準(zhǔn)，對(duì)各組以及再灌注損傷不同時(shí)間神經(jīng)功能損傷進(jìn)行神經(jīng)功能評(píng)分。觀察亞低溫對(duì)大鼠腦缺血再灌注損傷后神經(jīng)功能的影響；通過(guò)差速離心結(jié)合蛋白印跡分析各組大鼠全腦缺血再灌注24h后腦組織細(xì)胞中ASK1以及ASK1的磷酸化狀態(tài)和分布。 結(jié)果： 1.通過(guò)HE染色發(fā)現(xiàn)，亞低溫處理對(duì)大鼠全腦缺血再灌注后腦損傷起到保護(hù)作用，可減少神經(jīng)元的死亡。 2.通過(guò)改良的NSS神經(jīng)功能評(píng)分標(biāo)準(zhǔn)，證實(shí)了亞低溫處理出現(xiàn)全腦缺血再灌注后的大鼠，其NSS評(píng)分的分值能明顯降低，損傷的程度也明顯減輕。 3.通過(guò)Western Blot證實(shí)了在腦缺血再灌注24h后ASK1被活化，磷酸化后的ASK1呈現(xiàn)高表達(dá)狀態(tài)，亞低溫處理可明顯抑制ASK1的活化。在10min I/R組、亞低溫處理組均可測(cè)到活化的ASK1的高表達(dá)。相對(duì)于sham組，10min I/R組和亞低溫處理組中ASK1明顯被活化（pASK1），其表達(dá)性明顯增強(qiáng)，具有差異性，具有統(tǒng)計(jì)學(xué)意義（p0.05）。而相對(duì)于10min I/R組，亞低溫處理組中活化的ASK1的表達(dá)程度明顯低于10min I/R組中活化的ASK1，有差異性，，具有統(tǒng)計(jì)學(xué)意義（p0.05）。 結(jié)論： 1.亞低溫處理可對(duì)腦缺血再灌注后的神經(jīng)組織起保護(hù)作用，減少腦缺血再灌注后神經(jīng)細(xì)胞的死亡。 2.亞低溫處理可顯著減少腦缺血再灌注后神經(jīng)系統(tǒng)功能的損傷。 3.亞低溫處理對(duì)大鼠全腦缺血再灌注后神經(jīng)損傷的保護(hù)作用可能是通過(guò)抑制ASK1的活化，進(jìn)而抑制神經(jīng)細(xì)胞的死亡。
[Abstract]:Aim: to study the protective effect of mild hypothermia on global cerebral ischemia-reperfusion injury in rats and its mechanism. Methods: one hundred and fifty adult male Wistar rats were randomly divided into three groups: sham operation group (sham group), 10min ischemia / reperfusion group (10min I / R group) and mild hypothermia group. The whole brain ischemia reperfusion model was used in this experiment. The ischemia time was 10 min and the reperfusion time was 4 h / 24 h / 48 h and 72 h / h for 1 week. The rat hippocampal CA1 neurons after 24 hours of global cerebral ischemia and reperfusion were observed by high power microscope with HE staining and fixed with formaldehyde. The survival number of CA1 neurons in hippocampus was compared between each group. After the reperfusion time was reached, the nerve function of each group and the different time after reperfusion injury were evaluated by the modified NSS neural function scoring standard. To observe the effect of mild hypothermia on nerve function after cerebral ischemia-reperfusion injury in rats, the phosphorylation status and distribution of ASK1 and ASK1 in brain tissue cells of rats after 24 hours of global cerebral ischemia and reperfusion were analyzed by differential centrifugation and Western blotting. Results: 1. The results of HE staining showed that mild hypothermia treatment had a protective effect on brain injury after global cerebral ischemia reperfusion in rats and could reduce the death of neurons. 2. Through the improved NSS neural function scoring standard, it was proved that the NSS score of rats treated with mild hypothermia after global cerebral ischemia-reperfusion could be significantly reduced, and the degree of injury was also significantly reduced. 3. It was confirmed by Western Blot that ASK1 was activated 24 hours after cerebral ischemia-reperfusion, and the phosphorylated ASK1 showed high expression. Mild hypothermia significantly inhibited the activation of ASK1. High expression of activated ASK1 was detected in 10min I / R group and mild hypothermia group. Compared with sham group, 10min I / R group and mild hypothermia group were significantly activated (pASK1), the expression of ASK1 was significantly increased, with statistical significance (p0.05). Compared with 10min I / R group, the expression of activated ASK1 in mild hypothermia group was significantly lower than that in 10min I / R group (p0.05). Conclusion: 1. Mild hypothermia treatment can protect the nerve tissue after cerebral ischemia and reperfusion and reduce the nerve cell death after cerebral ischemia reperfusion. 2. Mild hypothermia treatment can significantly reduce the injury of nervous system function after cerebral ischemia reperfusion. 3. The protective effect of mild hypothermia on nerve injury after global cerebral ischemia-reperfusion in rats may be by inhibiting the activation of ASK1 and then inhibiting the death of nerve cells.
【學(xué)位授予單位】：吉林大學(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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