| 杨坤,胡锋,闵慧,梅求安,许明军.电针联合跑轮训练缓解大脑中动脉栓塞大鼠脑神经损伤和认知功能障碍的机制研究[J].中国康复,2023,38(2):67-72 |
| 电针联合跑轮训练缓解大脑中动脉栓塞大鼠脑神经损伤和认知功能障碍的机制研究 |
| Mechanism of electroacupuncture combined with running wheel training alleviating brain nerve injury and cognitive dysfunction in middle cerebral artery occlusion rats |
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| DOI： |
| 中文关键词: 电针联合跑轮训练 大脑中动脉栓塞（MCAO） Tau磷酸化 神经元 |
| 英文关键词: electroacupuncture combined with running wheel training middle cerebral artery occlusion Tau phosphorylation neurons |
| 基金项目:湖北省卫生健康委中医药科研项目（ZY2021M006） |
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| 中文摘要: |
| 目的：探究电针联合跑轮训练通过抑制tau过度磷酸化缓解大脑中动脉栓塞（MCAO）大鼠脑神经损伤的机制。方法：30只成年雄性SD大鼠被随机分为对照组（假手术处理，n=10）、MCAO组（进行MCAO手术，n=10），观察组（大鼠进行MCAO手术后的第7天，每天使用跑轮训练12h，持续10周，同时进行电针治疗）。采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法（TUNEL）染色分析大鼠脑组织中神经细胞凋亡情况；通过莫里斯水迷宫（MWM）测试大鼠的认知功能；采用蛋白印迹分析大鼠丘脑组织中凋亡蛋白Bcl2相关X蛋白（Bax）、B淋巴细胞瘤-2基因（Bcl-2）、B细胞淋巴瘤/白血病-xl基因（Bcl-xL）和陶蛋白（tau protein）蛋白的表达；采用尼氏染色和免疫荧光染色分析丘脑组织中神经元损伤情况；采用免疫组织化学检测大鼠海马中p-Tau丝氨酸396/404的磷酸化蛋白水平。结果：训练后，MCAO组脱氧核糖核苷酸末端转移酶介导的缺口末端标记法（TUNEL）染色阳性细胞凋亡率较对照组升高（P<0.05），观察组TUNEL染色阳性细胞凋亡率较MCAO组降低（P<0.05）。MCAO组发现隐藏平台的潜伏期较对照组增加（P<0.05），探测试验时间减少（P<0.05），观察组发现隐藏平台的潜伏期时间较MCAO组减少（P<0.05），探测试验时间增加（P<0.05）。MCAO组Bax蛋白表达较对照组升高（P<0.05），观察组Bax蛋白表达较MCAO组降低（P<0.05），MCAO组Bcl-2和Bcl-xL蛋白表达较对照组降低（P<0.05），观察组Bcl-2和Bcl-xL蛋白表达较MCAO组升高（P<0.05）。MCAO组完整神经元数量较对照组减少（P<0.05），观察组完整神经元数量较MCAO增多（P<0.05）。MCAO组p-Tauser396和p-Tause404磷酸化表达较对照组升高（P<0.05），观察组p-Tauser396和p-Tause404磷酸化表达较MCAO组降低（P<0.05）。MCAO组p-Tauser396、p-Tause404和总Tau蛋白表达升高（P<0.05），观察组p-Tauser396、p-Tause404和总Tau蛋白表达升降低（P<0.05）。结论：电针联合跑轮训练通过抑制tau过度磷酸化和tau蛋白表达减少MCAO大鼠脑损伤。 |
| 英文摘要: |
| Objective: To explore the mechanism of electroacupuncture combined with running wheel training alleviating brain nerve injury in middle cerebral artery occlusion (MCAO) rats by inhibiting Tau hyperphosphorylation. Methods: A total of 30 adult male SD rats were randomly divided into control group (sham operation, n=10), MCAO group (MCAO operation, n=10), and observation group (on the 7th day after MCAO operation, rats were treated with running wheel for 12 h a day for 10 weeks, and electroacupuncture was performed at the same time). TUNEL staining was used to analyze the apoptosis of nerve cells in rat brain. The cognitive function of rats was measured by Morris water maze（MWM）. Western blotting was used to analyze the expression of apoptotic proteins Bax, Bcl-2, Bcl-XL and Tau in rat thalamus tissue. Neuronal injury in thalamic tissue was analyzed by Nissner sta-ining and immunofluorescence staining. The levels of p-Tau ser396/404 phosphorylated protein in rat hippocampus were detected by immunohistochemistry. Results: The apoptosis rate of TUNEL staining positive cells in MCAO group was higher than that in control group (P<0.05), and that in observation group was lower than that in MCAO group (P<0.05). Compared with the control group, the latent period of the hidden platform in the MCAO group was increased (P<0.05) and the detection test time was decreased (P<0.05), while the latent period of the hidden platform in the observation group was decreased (P<0.05) and the detection test time was increased (P<0.05). The expression of Bax protein in MCAO group was higher than that in control group (P<0.05), the expression of Bax protein in observation group was lower than that in MCAO group (P<0.05), and the expression of Bcl-2 and Bcl-XL protein in MCAO group was lower than that in control group (P<0.05). The protein expression of Bcl-2 and Bcl-XL in observation group was higher than that in MCAO group (P<0.05). The number of intact neurons in MCAO group was less than that in control group (P<0.05), and that in observation group was greater than that in MCAO group (P<0.05). The phosphorylation expression of p-Tauser396 and p-Tause404 in MCAO group was higher than that in control group (P<0.05), and that of p-Tauser396 and p-Tause404 in observation group was lower than that in MCAO group (P<0.05). The expression levels of p-Tauser396, p-Tause404 and total Tau were increased in MCAO group (P<0.05), while the expression levels of p-Tauser396, p-Tause404 and total Tau were decreased in observation group (P<0.05). Conclusion: Electroacupuncture combined with running wheel training reduced cerebral nerve injury in MCAO rats by inhibiting Tau hyperphosphorylation and Tau protein expression. |
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