目的 探讨N-乙酰半胱氨酸（NAC）作为抗氧化剂在小鼠脑缺血再灌注后损伤的保护作用及其作用机制。 方法 小鼠随机分为假手术组、模型组和NAC组（150 mg/kg），每组18只。模型组和NAC组通过线栓法建立小鼠大脑中动脉缺血再灌注（MCAO）模型，NAC组小鼠于再灌注后立即腹腔注射NAC 150 mg/kg，假手术组和模型组小鼠给予等量0.9%氯化钠溶液。各组小鼠于再灌注24 h后进行神经行为学评分，使用激光散斑血流成像仪观察脑血流恢复情况，2,3,5-氯化三苯基四氮唑染色检测脑梗死体积，ELISA测定SOD和MDA含量， ROS荧光探针检测脑组织中ROS水平，免疫荧光法检测脑组织NeuN、Nrf2和HO-1蛋白表达及其与NeuN共定位情况，免疫印迹检测各组小鼠脑组织Nrf2和HO-1蛋白表达水平。 结果 NAC可令MCAO小鼠神经功能学评分降低，脑血流量增加，梗死体积减少，SOD水平升高且 MDA含量减少，NeuN阳性细胞率升高且ROS水平降低，Nrf2和HO-1蛋白表达水平增加（P<0.05），且Nrf2和HO-1与NeuN存在共定位。 结论 NAC可有效改善小鼠脑缺血再灌注引起的损伤，减少氧化应激，拯救损伤处神经元，其机制可能与调控Nrf2/HO-1信号通路有关。
Abstract
Objective To investigate the protective effects and mechanisms of N-acetylcysteine (NAC) as an antioxidant in ischemia-reperfusion injury in the mouse brain. Methods Mice were randomly divided into 3 groups: sham group, model group and NAC group (150 mg/kg), with 18 mice in each group. The model and NAC groups underwent middle cerebral artery occlusion (MCAO) followed by reperfusion, while the sham group underwent the same procedure without vessel occlusion. The NAC group received an intraperitoneal injection of NAC (150 mg/kg) immediately after reperfusion, whereas the sham and model groups received an equal volume of 0.9% sodium chloride solution. Neurological function was evaluated by behavioral tests 24 hours after reperfusion. Cerebral blood flow was assessed using laser speckle contrast imaging. Infarct volume was detected by 2,3,5-triphenyltetrazolium chloride staining. Superoxide dismutase (SOD) and malondialdehyde (MDA) levels were measured by ELISA. Reactive oxygen species (ROS) levels in brain tissue were detected using ROS fluorescent probes. The expression and colocalization of NeuN, Nrf2, and HO-1 proteins in brain tissue were examined by immunofluorescence. The protein levels of Nrf2 and HO-1 in brain tissue were analyzed by Western blot. Results NAC significantly reduced neurological function scores, increased cerebral blood flow, decreased infarct volume, elevated SOD levels, and reduced MDA content. The rate of NeuN-positive cells increased, and ROS levels decreased. The protein levels of Nrf2 and HO-1 increased (P<0.05), and Nrf2 and HO-1 colocalized with NeuN. Conclusions NAC effectively alleviates cerebral ischemia-reperfusion injury in mice, attenuates oxidative stress, and rescues damaged neurons, potentially through modulation of Nrf2/HO-1 signaling pathway.
关键词
N-乙酰半胱氨酸 /
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脑缺血再灌注 /
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氧化应激 /
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Nrf2/HO-1信号通路
Key words
N-acetylcysteine /
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Cerebral ischemia-reperfusion /
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Oxidative stress /
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Nrf2/HO-1 signaling
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基金
辽宁省科技厅应用基础项目(2023JH2/101700232)
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