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目的:探讨Peroxiredoxin 6(Prdx6)在脑缺血再灌注损伤中的影响及相关机制。方法:采用中脑动脉栓塞(middle cerebral artery occlusion,MCAO)模型诱导SD大鼠左侧脑皮质缺血再灌注,将大鼠随机(n=16)分为假手术组、MCAO组、Scramble组和Prdx6-si RNA组:前两组均不进行侧脑室注射,其中Sham组仅做手术切口,不插入线栓,MCAO组进行手术造模;后2组分别侧脑室注射8μl的乱序Prdx6-si RNA和Prdx6-si RNA再手术建模。测定各组神经功能学评分、脑含水量和脑梗死体积;HE和亚甲蓝染色法检测大脑皮质缺血再灌注区神经元的损伤情况;免疫印迹法检测脑组织中Prdx6及Caspase3蛋白的表达水平;最后,检测SOD、MDA活性并用TUNLE染色法探讨相关氧化、凋亡机制。结果:Prdx6-si RNA组与MCAO组相比,神经功能学评分、缺血侧脑含水量和脑梗死体积均明显增加(P=0.000);HE和亚甲蓝染色显示脑缺血区神经元损伤进一步加重(P=0.000);SOD活性进一步降低(P=0.000),MDA含量进一步增加(P=0.000);TUNLE染色阳性细胞率(P=0.000)和Caspase3蛋白表达水平(P<0.01)均进一步增高。而Scramble组与MCAO组比以上各指标均无明显统计学差异。结论:Prdx6在脑缺血再灌注损伤中起保护作用,此作用与其抗氧化和抗凋亡机制有关。
Objective: To investigate the effect of Peroxiredoxin 6 (Prdx6) on cerebral ischemia-reperfusion injury and its related mechanisms. Methods: The middle cerebral artery occlusion (MCAO) model was used to induce left cerebral cortex ischemia-reperfusion in rats. The rats were randomly divided into sham operation group, MCAO group, Scramble group and control group Prdx6-si RNA group: The first two groups were not intracerebroventricular injection, which Sham group only surgical incision, do not insert the thread plug, MCAO group for surgical modeling; the latter two groups were injected into the lateral ventricle 8μl of disorder Prdx6-si Reconstruction of RNA and Prdx6-si RNAs. Neurological scores, brain water content and infarct volume were measured in each group. Neurons injury in cerebral ischemia-reperfusion area was detected by HE and methylene blue staining. Expression of Prdx6 and Caspase3 protein in brain tissue was detected by Western blotting Finally, the activities of SOD and MDA were detected and the related mechanisms of oxidation and apoptosis were investigated by TUNLE staining. Results: Compared with MCAO group, Prdx6-si RNA significantly increased neurological score, water content in cerebral ischemia and volume of cerebral infarction (P = 0.000). HE and methylene blue staining showed that neurons in cerebral ischemic area (P = 0.000); the activity of SOD was further decreased (P = 0.000); MDA content was further increased (P = 0.000); the positive rate of TUNLE staining (P = 0.000) and the expression of Caspase3 protein Further increase. The Scramble group and MCAO group than the above indicators were no significant difference. Conclusion: Prdx6 plays a protective role in cerebral ischemia-reperfusion injury, and its role is related to its anti-oxidation and anti-apoptotic mechanisms.