| 张贤,张保,柏中喜,刘迪,储小宁,郑苏.经颅直流电刺激联合穴位针刺对脑梗死大鼠学习记忆能力、神经细胞自噬及脑组织BDNF、NGF表达的影响[J].中国康复,2025,40(5):259-264 |
| 经颅直流电刺激联合穴位针刺对脑梗死大鼠学习记忆能力、神经细胞自噬及脑组织BDNF、NGF表达的影响 |
| Effects of transcranial direct current stimulation (tDCS) combined with acupoint acupuncture on learning and memory ability, nerve cell autophagy, and the expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in the brain tissue of ischemic stroke rats |
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| DOI：10.3870/zgkf.2025.05.001 |
| 中文关键词: 脑梗死 经颅直流电刺激 穴位针刺 学习记忆 神经细胞自噬 脑源性神经营养因子 脑神经生长因子 |
| 英文关键词: ischemic stroke transcranial direct-current stimulation acupoint acupuncture learning and memory nerve cell autophagy brain-derived neurotrophic factor nerve growth factor |
| 基金项目:中医药科研立项项目（青年人才项目）（ZY2021Q015） |
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| 目的：探究经颅直流电刺激（tDCS）联合穴位针刺对脑梗死大鼠学习记忆能力、神经细胞自噬及脑源性神经营养因子（BDNF）、脑神经生长因子（NGF）表达的影响。方法：选取健康雄性无特定病原体（SPF）级SD大鼠45只，将其中35只大鼠建立脑梗死模型，本次建模成功30只，随机分为脑梗死组、tDCS组、联合组，10只/组，另将剩余10只健康大鼠作为对照组。其中对照组与脑梗死组不进行干预；tDCS组给与tDCS治疗，1次/d，共14d；联合组在tDCS组基础上联合穴位针刺干预，1次/d，共14d。采用水迷宫实验检测大鼠学习记忆能力，苏木精-伊红染色法（HE）观察大鼠脑组织病理改变，免疫组化法检测大鼠脑组织BDNF、NGF表达，免疫印迹检测大鼠大脑海马CA1区神经细胞自噬相关蛋白(Beclin1)、微管相关蛋白1轻链3(LC3)表达，逆转录聚合酶链式反应(RT-PCR)法检测大鼠大脑海马CA1区Beclin1、LC3 mRNA水平。结果：与对照组相比，脑梗死组大鼠的逃避潜伏期延长、定向角增大、Beclin1及LC3 mPNA表达水平升高，第3象限停留时间 (RTQ)缩短、跨平台次数减少、海马BDNF及NGF含量降低（P＜0.05）；与脑梗死组相比，tDCS组大鼠的逃避潜伏期缩短、定向角减小、Beclin1及LC3 mPNA表达水平降低，RTQ延长、跨平台次数增加、海马BDNF及NGF含量升高（P＜0.05）；与 tDCS 组相比，联合组大鼠的逃避潜伏期缩短、定向角减小、Beclin1及LC3 mPNA表达水平降低，RTQ延长、跨平台次数增加、海马BDNF及NGF含量升高（P＜0.05）。HE 染色结果显示，对照组大鼠脑组织结构完好，脑梗死组大鼠脑组织结构破坏严重，出现大量细胞排列紊乱及空泡变性，tDCS 组与联合组病理状态均有所减轻，神经细胞核及核仁形态逐渐清晰完整，其中以联合组表现最为明显。结论：tDCS 联合穴位针刺对治疗脑梗死具有良好效果，其机制可能与增强学习记忆能力、改善神经细胞自噬及调节BDNF、NGF水平相关。 |
| 英文摘要: |
| Objective: To explore the impacts of transcranial direct-current stimulation (tDCS) combined with acupoint acupuncture on the learning and memory ability, nerve cell autophagy, and the expression levels of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in ischemic stroke rats. Methods: In total, 45 healthy male SPF SD rats were selected. Among them, 35 rats were used to establish a stroke model. Totally, 30 rats were successfully modeled and then divided into the ischemic stroke group, the tDCS group, and the combined group, with 10 rats in each group. The remaining 10 rats served as the control group. The control group and the ischemic stroke group received no intervention. The tDCS group was treated with tDCS once a day for 14 days. The combined group received acupoint acupuncture intervention in addition to tDCS treatment once a day for 14 days. The Morris water maze test was employed to assess the learning and memory ability. Pathological changes were ob-served by hematoxylin-eosin (HE) staining. The expression of BDNF and NGF was detected by immunohistochemistry. Western blotting was used to detect the nerve cell autophagy-related proteins Beclin1 and LC3, and reverse-transcription polymerase chain reaction (RT-PCR) was applied to detect Beclin1 and LC3 mR-NA. Results: Compared with the control group, the escape latency, orientation angle, Beclin1 and LC3 protein expression level increased, while the residence time in the third quadrant (RTQ), cross-platform frequency, and BDNF and NGF content decreased in the ischemic stroke group (P<0.05). Compared with the ischemic stroke group, the escape latency, orientation angle, Beclin1 and LC3 protein expression level decreased, while the RTQ, cross-platform frequency, and BDNF and NGF content increased in the tDCS group (P<0.05). Compared with the tDCS group, the escape latency, orientation angle，Beclin1 and LC3 protein expression level further decreased, while the RTQ, cross-platform frequency, and BDNF and NGF content further increased in the combined group (P<0.05). HE staining results demonstrated that the brain tissue of rats in the control group was intact, while that in the ischemic stroke group was severely damaged, with a large number of cells showing disordered arrangement and vacuolar degeneration. The pathological conditions in both the tDCS group and the combined group were alleviated, and the morphology of nerve cell nuclei and nucleoli gradually became clear and complete, with the combined group showing the most significant improvement. Conclusion: The tDCS combined with acupoint acupuncture exerts a favorable effect on the treatment of stroke. The mechanism may be associated with enhancing learning and memory ability, improving nerve cell autophagy, and regulating the levels of BDNF and NGF. |
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