头穴丛刺法调控大鼠脑梗死后内源性神经干细胞增殖迁移分化的实验研究

关键词：脑梗死  头穴丛刺法  室管膜下区  神经干细胞

Keywords:cerebral infarction  scalp clustering acupuncture  subventricular zone  neural stem cell

摘要：

      目的：本实验通过观察头穴丛刺法对室管膜下区(SVZ)的神经干细胞(NSC)增殖、迁移、分化的调控作用，探讨该疗法促进成年神经再生、修复和重建神经网络的理论机制。方法： 大鼠注射荧光染料Dil以预标记室SVZ细胞；栓线法制备大鼠局灶性脑缺血模型。将造模成功的Wistar大鼠随机分成4组：模型组12只，头针组12只，头针丛刺法组12只，假手术组4只。采用脉冲式的BrdU标记方法标记新生细胞；采用激光共聚焦显微镜检测预标记原始SVZ NSC后，再检测双重免疫荧光染色所确定的分化的细胞。结果：①模型组、头针组、头穴丛刺法组均可见Dil标记的SVZ细胞迁移至梗死周边的纹状体和皮质，并且分化成神经元或胶质细胞，头穴丛刺法组Dil/BrdU/NeuN或Dil/BrdU/GFAP标记的细胞明显增多，与其他两组比较有显著性差异（P<0.01）；②头穴丛刺法组BrdU/NeuN及BrdU/GFAP阳性细胞表达均多于其他两组，组间比较有显著性差异（P<0.01）。结论：①头穴丛刺法能促进脑缺血后SVZ 区神经干细胞增殖，并且随时间递增减少其增殖衰减；②此法可促进Dil标记的SVZ细胞均迁移至梗死周边的纹状体和皮质，并且分化成神经元或胶质细胞；③此法能促进局灶性脑缺血后SVZ NSC的迁移和分化。

Abstract:

      Objective: To observe the regulation effect of scalp clustering acupuncture therapy on the proliferation, migration and differentiation of neural stem cells(NSC) in subventricular zone(SVZ) and explore the therapeutic effect and mechanism of promoting adult neurogenesis, repair and reconstitution of neural networks. Method: Stereotaxic apparatus was used to locate the coordination in rats. Dilstain(Dil) was injected into the lateral cerebrum ventricle for prelabeling the SVZ cells. The cerebral ischemia model of rat was made by blocking middle cerebral artery. Rats were randomly divided into four groups: A-sham operation group(4 rats),R-model group(12 rats), C-scalp acupuncture group(12 rats), D-scalp clustering acupuncture group(12 rats). Pulsed BrdU method was used to label new cells. NSCs in SVZ prelabeled with Dil were observed with laser confocal microscopy, and double immunofluorescent staining was used to identify cells differentiation. Result: ①Dil label cells in B, C, D groups all migrated into peri-infarct regions and differentiated into neurons and astrocytes. Dil/BrdU/NeuN or Dil/BrdU/GFAP－positive cells in D group increased significantly, compared with B and C groups(P<0.01); ②Expressions of BrdU/NeuN and BrdU/GFAP-positive cells in D group increased significantly compared with B and C groups(P<0.01).Conclusion: ①The scalp clustering acupuncture therapy can promote NSCs proliferation in neuroproliferation center- SVZ after focal ischemia, to reduce the diminishment of proliferation with the time passing. ②This therapy can promote Dil-label cells in SVZ migrating to striatum and cortex in peri-infarct region and differentiating into neurons and astrocytes. ③This therapy can promote the migration of SVZ NSCs after focal ischemia and the differentiation of endogenous NSCs.