目的: 探究褪黑素对卒中后认知障碍的保护作用及机制。方法: 雄性C57BL/6J小鼠被随机分为假手术组、卒中组、卒中+褪黑素组（n=7/组）,采用光化学栓塞法构建缺血性脑卒中模型,造模后连续21 d腹腔注射生理盐水或褪黑素（20 mg/kg）,行为学检测小鼠的认知功能、尼氏染色检测神经元数量、Western blotting检测凋亡、突触可塑性和线粒体自噬相关蛋白的水平。结果: 与假手术小鼠相比,卒中小鼠在新物体识别实验中的分辨指数显著降低、Y迷宫中新异臂的停留时间显著减少;海马和皮层神经元数量显著减少;促凋亡蛋白（Bax 、cleaved Caspase-3）水平上调、突触相关蛋白（PSD-95、Synaptophysin）水平下调、线粒体自噬相关蛋白（PINK1、Parkin）水平上调。褪黑素治疗显著提高了卒中小鼠在新物体识别实验中的分辨指数、增加了Y迷宫中新异臂的停留时间;海马和皮层神经元数量显著增多;促凋亡蛋白水平下调、突触相关蛋白水平上调、线粒体自噬相关蛋白水平下调。结论: 褪黑素抑制了过度的线粒体自噬并改善了卒中后认知障碍。
Abstract
Objective This study aimed to investigate the protective effect and mechanism of melatonin (MT) in post-stroke cognitive impairment (PSCI). Methods Male C57BL/6J mice were randomly divided into Sham, Stroke, and Stroke+MT groups (n=7/group). Photothrombotic-induced stroke model mice were injected intraperitoneally with saline or MT (20 mg/kg) for 21 consecutive days after modeling. Behavioral tests were performed to examine the cognitive function of the mice, Nissl staining to detect neuronal damage, and Western blotting to detect apoptosis, synaptic plasticity and mitophagy. Results Compared with Sham mice, stroke mice showed a significantly reduction in the discrimination index in the novel object recognition test and the time spent in the novel arm of the Y maze. The number of neurons in the hippocampus and cortex were significantly reduced. The levels of proapoptotic proteins (Bax, cleaved Caspase-3) were upregulated, the levels of synapse-related proteins (PSD-95, Synaptophysin) were downregulated, and the levels of mitophagy-related proteins (PINK1, Parkin) were upregulated. MT significantly increased the discrimination index in the novel object recognition test and the time spent in the novel arm of the Y maze. The number of hippocampal and cortical neurons were significantly increased. The levels of proapoptotic proteins (Bax, cleaved Caspase-3) were downregulated, the levels of synapse-related proteins (PSD-95, Synaptophysin) were upregulated, and the levels of mitophagy-related proteins (PINK1, Parkin) were downregulated. Conclusion MT inhibited excessive mitophagy and ameliorated PSCI.
关键词
卒中后认知障碍 /
线粒体自噬 /
褪黑素 /
海马体 /
凋亡
Key words
post-stroke cognitive impairment /
mitophagy /
melatonin /
hippocampus /
apoptosis
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基金
湖南省卫生健康委科研项目“空间学习改善大鼠抑郁样行为的机制研究”（202102061699）; 湖南师范大学本科生创新创业计划项目“褪黑素改善卒中后认知障碍的机制研究”（2023289）
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