[目的]观察通督调神针法对大脑中动脉闭塞(MCAO)模型大鼠行为学、缺血半暗带超微结构和突触体相关蛋白25(SNAP-25)的影响,探析电针疗法的神经保护机制.[方法]将40只SD大鼠随机分为空白组、假手术组、模型组和电针组,每组10只.空白组不做干预,假手术组大鼠仅接受皮肤切开及血管剥离术,模型组及电针组大鼠采用线栓法进行脑缺血模型制备.造模成功后电针组予通督调神针法干预,1次/日,每次30 min,连续干预7d.空白组、假手术组及模型组术后在同等条件下抓取,回笼饲养.记录各组大鼠神经功能评分,采用2,3,5氯化三苯基四氮唑(TTC)染色法检测大鼠脑梗死面积,透射电镜观察大鼠脑组织缺血半暗带处超微结构,Western blot 检测各组大鼠海马区SNAP-25蛋白表达水平.[结果]模型组大鼠神经功能评分明显低于假手术组和空白组(P<0.05);与模型组比较,电针组大鼠神经功能评分显著增加,差异有统计学意义(P<0.05).模型组和电针组大鼠存在明显脑梗死区域,电针组脑梗死面积小于模型组(P<0.01).空白组和假手术组大鼠脑组织神经元细胞结构完整,细胞核膜完整,结构清晰,整体细胞器完整;模型组大鼠脑组织神经元细胞结构受破坏,包括细胞膜不完整、细胞核肿胀、内质网减少等;与模型组比较,电针组大鼠脑组织具有较为完整的细胞膜及细胞核结构,线粒体空泡现象较少.与空白组、假手术组比较,模型组大鼠缺血半暗带脑组织的SNAP-25蛋白表达水平明显增加(P<0.01);与模型组比较,电针组大鼠缺血半暗带脑组织的SNAP-25蛋白表达水平明显减少(P<0.01).[结论]通督调神针法可通过下调SNAP-25蛋白表达水平,抑制神经突触小体增殖及修复神经细胞结构,从而发挥神经保护作用.
Effects of Tongdu Tiaoshen Electroacupuncture on Ultrastructure of Ischemic Penumbra and SNAP-25 Protein in MCAO Rats
[Objective]To observe of the effects of Tongdu Tiaoshen Electroacupuncture(EA)on ethology,ultra-structure of ischemic semi-dark band and synaptosomal-associated protein 25(SNAP-25)in rats with middle cerebral artery occlusion(MCAO)model and explore the neuroprotective mechanism of Tongdu Tiaoshen EA.[Methods]40 SD rats were randomly divided into blank group,sham-operated group,model group and EA group,with 10 in each group.Animals in the blank group received no intervention.Animals in the sham-operated group were received skin incision and vascular stripping only,while rats in the model group and the EA group received operation for cerebral ischemia modeling with suture-occluded method.After successful modeling,the EA group was given Tongdu Tiaoshen Elec-troacupuncture,1 time a day,30 min each time,for 7 consecutive days.Rats in the blank,sham-operated and model groups were captured,caged and fed under equal conditions.The neurological function scores in all groups were record-ed.The area of cerebral infarction was measured by 2,3,5 triphenyltetrazolium chloride(TTC)staining.The ultrastruc-ture of the ischemic penumbra in the brain tissues was observed by transmission electron microscopy.And the level of SNAP-25 protein expression in hippocampus was measured by Western blot.[Results]The nerve function score in the model group was significantly lower than that in the sham-operated group and the blank group(P<0.05).And the score in the EA group was significantly higher than that of the model group(P<0.05).Cerebral infarction areas were found in both the model group and the EA group.But the infarction areas in the EA group were significantly smaller than that in the model group(P<0.01).The neuron cells in rat brain tissue in the blank and sham-operated groups remained integri-ty,with intact nuclear membrane,well-defined structure,and intact organelles.The neuron cell structure of brain tissue in the model group was damaged,including incomplete cell membranes,swollen nuclei,and reduced endoplasmic retic-ulum.Compared with the model group,the brain tissue of rats in the EA group had a relatively complete cell membrane and nucleus structure,and the mitochondrial vacuole was significantly reduced.Compared with the blank group and the sham-operated group,the SNAP-25 protein expression in the ischemic penumbra in the model group was significantly increased(P<0.01).Compared with the model group,the SNAP-25 protein expression in the ischemic penumbra in the EA group was significantly reduced(P<0.01).[Conclusion]The Tongdu Tiaoshen EA can inhibit the proliferation of synaptosomes and repair the structure of neuron by reducing the expression of SNAP-25 protein,thus exerting a neuro-protective effect.
Tongdu Tiaoshen Electroacupuncturemiddle cerebral artery occlusioncerebral ischemic injuryul-trastructureSNAP-25experimental research
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