秦芳,马甜甜,于子夫,等.针刺同步运动训练对大脑中动脉闭塞大鼠学习记忆能力的影响及机制研究[J].中华物理医学与康复杂志,2025,47(7):577-584
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| 针刺同步运动训练对大脑中动脉闭塞大鼠学习记忆能力的影响及机制研究 |
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| DOI：10.3760/cma.j.cn421666-20241105-00885 |
| 中文关键词: 大脑中动脉闭塞 针刺同步运动训练 线粒体 自噬 |
| 英文关键词: Middle cerebral artery occlusion Acupuncture-synchronized exercise Mitochondria Autophagy |
| 基金项目:国家自然科学基金青年基金项目（81802239）；2022年度山东省医务职工科技创新计划立项项目（SDYWZGKCJH2022024） |
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| 中文摘要: |
| 目的 观察针刺同步运动训练对大脑中动脉闭塞（MCAO）大鼠学习记忆能力的影响，并探讨相关机制。 方法 选取60只8周龄无特定病原体（SPF）级SD雄性大鼠，随机抽取6只纳入假手术组，其余大鼠构建MCAO模型。将造模成功的18只大鼠按照随机数字表法分为模型组、非同步组、同步组，每组6只。假手术组、模型组不予特殊干预，非同步组先进行针刺干预20 min、取针后进行跑台训练20 min，同步组在针刺留针的同时进行跑台训练20 min，每日1次，每周5 d，共21 d。造模术后24 h、干预21 d后，采用Longa评分法评估大鼠的神经功能缺损程度。干预第17～20天进行定位航行试验并记录大鼠的逃避潜伏期，干预第21天进行空间探索实验并检测大鼠在原平台象限的停留时间和穿越次数。干预21 d后，采用苏木精-伊红（HE）染色法观察大鼠海马组织的神经元形态，利用透射电镜观察大鼠海马神经元线粒体情况，并检测丙二醛（MDA）、超氧化物歧化酶（SOD）水平。采用Western blot法检测Parkin、PINK1、LC3-Ⅱ/ LC3-Ⅰ、P62、Beclin-1蛋白表达水平。 结果 与假手术组比较，模型组大鼠的Longa评分较高（P<0.05），目标象限停留时间［(14.51±2.99)s］和穿越平台次数［(1.83±1.17)次］减少（P<0.05），HE染色显示存在大量变性神经元，线粒体肿胀呈空泡化、嵴稀疏，Parkin、PINK1、LC3-Ⅱ/ LC3-Ⅰ、Beclin-1及SOD水平均显著降低（P<0.05），P62和MDA水平显著升高（P<0.05）。与模型组比较，非同步组和同步组大鼠的Longa评分降低（P<0.05），目标象限停留时间和穿越平台次数增加（P<0.05），HE染色显示神经元的形态接近正常，线粒体嵴结构较完整、有自噬小体，Parkin、PINK1、LC3-Ⅱ/ LC3-Ⅰ、Beclin-1及SOD水平均升高（P<0.05），P62和MDA水平降低（P<0.05）。与非同步组比较，同步组大鼠目标象限停留时间［(28.10±5.30)s］和穿越平台次数［(5.33±1.03)次］增加（P<0.05）， LC3-Ⅱ/ LC3-Ⅰ、Beclin-1和SOD水平升高（P<0.05），P62和MDA水平降低（P<0.05）。 结论 针刺同步运动训练可以改善MCAO大鼠的学习记忆能力，其机制可能与降低氧化应激水平、激活线粒体自噬有关。 |
| 英文摘要: |
| Objective To observe the effect of acupuncture-synchronized exercise training on the learning and memory of rats after middle cerebral artery occlusion (MCAO), and to explore its mechanism. Methods Among sixty 8-week-old SPF-grade Sprague-Dawley male rats, 6 were randomly chosen to form a sham group, and the rest received MCAO. After successful modeling, 18 rats were assigned at random to either the model, asynchronous or synchronous group, each of 6. The asynchronous group received 20 minutes of acupuncture followed by 20 minutes of treadmill training, while the synchronous group received the acupuncture during their treadmill training. There was no special intervention for the sham and model groups. There were 5 sessions/week for 21 days. Neurological deficit in the rats was assessed using Longa scoring 24h after the modeling and after the 21 days. The localization navigation test was performed on days 17-20 of the experiment, and the latency to escape was recorded. A space exploration experiment was performed the next day, and rats were tested for dwell time and the number of transits of the original platform quadrant. After the intervention, the morphology of hippocampal tissue was observed using HE staining. The mitochondria of the rats′ hippocampal neurons were observed using transmission electron microscopy. MDA and SOD levels were recorded. And the protein expression levels of the Parkin, PINK1, LC3-II/ LC3-Ⅰ, P62 and Beclin-1 genes were detected using western blotting. Results The model group showed significantly higher average Longa scores than the sham group, with less time spent in the target quadrant and fewer crossings of the platform quadrant. The HE staining revealed numerous degenerating neurons, and swollen mitochondria with vacuolization and sparse cristae. There was also a significant decrease in Parkin, PINK1, LC3-II/LC3-I and Beclin-1 expression and in SOD. But there was a significant increase in P62 expression and MDA, on average. Compared with the model group, both the non-synchronous and synchronous groups showed significantly lower average Longa scores, with more time spent in the target quadrant and more platform crossings. Their neuron morphology was close to normal, with more intact cristae and autophagic bodies in the mitochondria. Those groups showed greater Parkin, PINK1, LC3-II/LC3-I and Beclin-1 expression and had higher SOD levels, on average, with significantly lower P62 expression and less MDA. But compared with the non-synchronous group, the synchronous group spent more time in the target quadrant and had significantly more platform crossings. LC3-II/LC3-I and Beclin-1 expression were higher, on average, as were SOD levels. P62 expression and MDA levels were significantly lower. Conclusions Acupuncture synchronized with exercise improves the learning and memory of rates after MCAO. The mechanism may be related to reducing oxidative stress levels and activating mitochondrial autophagy. |
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