南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (5): 679-686.doi: 10.12122/j.issn.1673-4254.2021.05.07

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DJ-1通过Nrf2信号通路减轻大鼠脑缺血-再灌注引起的氧化应激损伤

李 莉，彭 莉，朱 进，巫静娴，赵 涌   

  • 出版日期:2021-05-20 发布日期:2021-06-11

DJ-1 alleviates oxidative stress injury by activating the Nrf2 pathway in rats with cerebral ischemia-reperfusion injury

  • Online:2021-05-20 Published:2021-06-11

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摘要： 目的 探讨DJ-1（Park7）在脑缺血再灌注损伤中的抗氧化作用及相关机制。方法 构建SD大鼠大脑缺血再灌注损伤模型，将SD大鼠随机分为假手术组、MCAO 组、Scramble组和 DJ-1 siRNA组、NC组和 Overexpression组。通过DJ-1 siRNA干扰片段干扰DJ-1的表达，以及DJ-1过表达腺相关病毒载体过表达DJ-1的表达。通过测定MCAO/R后各组神经功能学评分和脑含水量，同时应用HE和Nissl 染色法评估脑组织的形态学变化和皮层梗死区神经元的损伤情况，评价 DJ-1 对大鼠脑缺血再灌注损伤的神经保护作用。用超氧化物歧化酶（SOD）和丙二醛（MDA）分析脑组织氧化应激状态。免疫印迹法检测脑组织中 DJ-1，Nrf2，HO-1以及NQO1的表达水平，以及免疫荧光染色法观察Nrf2的表达及核转位情况，探讨DJ-1减轻大鼠脑缺血再灌注氧化应激损伤的可能机制。结果 与 MCAO 组相比，DJ-1 siRNA 组的神经功能学评分（P?0.001）、脑含水量（P?0.001）均明显增加；HE 和Nissl 染色显示脑缺血区神经元损伤进一步加重；SOD 含量进一步降低，MDA 含量进一步增加（P?0.001）；干扰DJ-1后，其蛋白水平明显降低（P=0.003），同时Nrf2及其下游的HO-1和NQO1也明显降低（P?0.001）。而过表达DJ-1以后，其DJ-1 （P=0.006）、Nrf2（P=0.006）及其下游的HO-1（P=0.004）和NQO1明显增加（P=0.014）。结论 DJ-1作为体内重要的神经保护因子，可以减少大鼠脑缺血再灌注氧化应激损伤，并可能是通过激活Nrf2信号通路来实现的。

关键词: DJ-1；Nrf2信号通路；脑缺血-再灌注；氧化应激

Abstract: Objective To investigate the antioxidant effect of DJ-1 (Park7) in rats with cerebral ischemia/reperfusion (IR) injury and its potential mechanism. Methods A total of 108 SD rats were randomly divided into sham-operated group, middle cerebral artery occlusion (MCAO) group, Scramble group, DJ-1 siRNA group, negative control (NC) group and DJ-1 overexpression group. Except for those in the sham group, all the rats were subjected to MCAO to establish models of cerebral IR injury. In DJ-1 siRNA and DJ-1 overexpression group, a DJ-1 siRNA and an adeno-associated virus vector carrying DJ-1 gene was injected into the lateral ventricle of the rats, respectively. In each group, neurological scores and brain water content were determined after the operation, and pathological changes of the brain tissue and neuronal injury in the cortical infarction area were assessed using HE and Nissl staining. Oxidative stress in the brain tissues was analyzed by detecting superoxide dismutase (SOD) and malondialdehyde (MDA). The expression levels of DJ-1, Nrf2, Ho-1 and NQO1 in the brain tissue were detected with Western blotting, and the expression and nucleation of Nrf2 was determined by immunofluorescence staining. Results Compared with those in MCAO group, the neurological scores (P<0.001) and brain water content (P<0.001) were significantly increased in DJ-1 siRNA group. Intracerebral injection of DJ-1 siRNA following MCAO obviously aggravated neuron injury in cerebral ischemia region, further reduced SOD activity and increased MDA content (P<0.001), and significantly lowered the expression levels of Nrf2 and its downstream proteins HO-1 and NQO1 (P<0.001). Intracerebral injection of the adenoviral vector for DJ-1 (P=0.003) overexpression significantly upregulated the levels of Nrf2 (P=0.006) and its downstream proteins HO-1 (P=0.004) and NQO1 (P=0.014). Conclusion As an important neuroprotective factor, DJ-1 alleviates oxidative stress induced by cerebral IR injury in rats by activating the Nrf2 pathway.

Key words: DJ-1; Nrf2 pathway; cerebral ischemia-reperfusion; oxidative stress