亚低温对大鼠全脑缺血再灌注性脑损伤的保护作用及机制研究
发布时间：2019-02-23 10:44
【摘要】：目的： 研究亚低温对大鼠全脑缺血再灌注性脑损伤的保护作用，并初步探讨其机制。 方法： 选取成年雄性Wistar大鼠150只，实验大鼠随机分为3组，每组50只，即分为假手术组（sham组）、10min缺血/再灌注组（10min I/R组）、亚低温处理组。本实验采用大鼠全脑缺血再灌注模型，缺血时间为10min，再灌注时间为4h，24h，48h，72h，1周。通过甲醛固定脑组织并切片，通过HE染色运用高倍显微镜观察全脑缺血再灌注24h后的大鼠海马CA1区神经元细胞，比较各组间大鼠海马CA1区神经元存活数量；达到再灌注时间后，通过改良的NSS神经功能评分标准，对各组以及再灌注损伤不同时间神经功能损伤进行神经功能评分。观察亚低温对大鼠脑缺血再灌注损伤后神经功能的影响；通过差速离心结合蛋白印迹分析各组大鼠全脑缺血再灌注24h后脑组织细胞中ASK1以及ASK1的磷酸化状态和分布。 结果： 1.通过HE染色发现，亚低温处理对大鼠全脑缺血再灌注后脑损伤起到保护作用，可减少神经元的死亡。 2.通过改良的NSS神经功能评分标准，证实了亚低温处理出现全脑缺血再灌注后的大鼠，其NSS评分的分值能明显降低，损伤的程度也明显减轻。 3.通过Western Blot证实了在脑缺血再灌注24h后ASK1被活化，磷酸化后的ASK1呈现高表达状态，亚低温处理可明显抑制ASK1的活化。在10min I/R组、亚低温处理组均可测到活化的ASK1的高表达。相对于sham组，10min I/R组和亚低温处理组中ASK1明显被活化（pASK1），其表达性明显增强，具有差异性，具有统计学意义（p0.05）。而相对于10min I/R组，亚低温处理组中活化的ASK1的表达程度明显低于10min I/R组中活化的ASK1，有差异性，，具有统计学意义（p0.05）。 结论： 1.亚低温处理可对脑缺血再灌注后的神经组织起保护作用，减少脑缺血再灌注后神经细胞的死亡。 2.亚低温处理可显著减少脑缺血再灌注后神经系统功能的损伤。 3.亚低温处理对大鼠全脑缺血再灌注后神经损伤的保护作用可能是通过抑制ASK1的活化，进而抑制神经细胞的死亡。
[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.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.3
本文编号：2428743
[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.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.3
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