头穴丛刺法对脑缺血大鼠半暗带区突触素SYN表达影响的研究

发布时间：2018-09-07 19:01

【摘要】：缺血性脑卒中是脑卒中类疾病中最常见的类型,其发生过程具有复杂的生理、病理机制。近年来,神经血管单元(NVU)概念的提出为缺血性脑卒中的病理机制及治疗策略的研究开辟了新的方向,针对NVU中各结构损伤的多靶向治疗方案将对脑卒中后的神经功能恢复及整体预后有着更好的影响。头穴从刺法是于致顺教授根据基础理论体系,再结合实践中独到的临床经验总结出来的一种头针治疗方法,临床上治疗缺血性卒中疗效确切,根据其“场”假说,头穴丛刺法对脑组织具有广泛的调节作用,头穴丛刺法治疗缺血性卒中的作用机理可能与促进神经血管单元结构及功能恢复有关。神经元是NVU的组成部分之一并在其中占主导地位,突触是神经元间或神经元与效应器细胞间起神经信号传导作用的部位,对NVU各组分间的相互联系和影响有着重要意义。突触素(synapsin, SYN)是一种钙结合蛋白,其特异性分布于突触囊泡膜上,缺血性脑卒中后SYN表达量的回升标志着突触结构及功能的重建,亦标志着NVU中神经元功能的恢复。为了验证头穴丛刺法在缺血性脑卒中的治疗作用机制,本研究设计了头穴丛刺法对大鼠半暗带区突触素SYN表达影响的动物实验课题。目的：观察头穴丛刺法对脑缺血大鼠神经功能评分和半暗带区突触素SYN阳性表达面积的影响,研究头穴丛刺法对缺血性脑卒中后神经功能恢复作用机制。方法：将54只雄性SD大鼠平均分成头穴丛刺组(A组),模型组(B组),假手术组(C组),每组18只大鼠。每组按3个时间点(1天、7天、14天)随机平均分为3个子小组,每时间点每小组6只大鼠。A、B组采用改良线栓法制备MCAO模型,C组仅进行假手术操作。A组大鼠采用头穴丛刺法,B与C组不予任何治疗干预。按Bederson法对各组大鼠造模后24h及取材前1h进行神经功能评分；取材后,大鼠脑组织切片以免疫荧光染色,用荧光显微镜寻找半暗带区,拍摄大鼠脑组织半暗带区SYN表达情况,使用生物图像分析软件计算SYN阳性表达面积百分比。结果：1.A与B组比较,7天及14天时神经功能评分均降低,差异具有统计学意义(P0.05)：A组造模后与取材前比较,7天及14天时神经功能评分降低(P0.05)；B组造模后与取材前比较,14天时神经功能评分降低(P0.05)；A组与B组比较,7天及14天时,大鼠脑组织半暗带区突触素SYN表达增加(P0.05)；A组大鼠脑组织半暗带区SYN在1天、7天、14天时表达量逐渐增加(P0.05);结论：1.MCAO模型可造成大鼠神经功能缺损,降低半暗带区突触素SYN表达；2.头穴丛刺法可持续改善脑缺血大鼠神经功能评分,对神经功能的恢复有着积极的影响；3.头穴丛刺法能明显增加脑缺血大鼠半暗带区突触素SYN表达水平,促进NVU中神经元突触结构及功能重建；4.大鼠脑缺血后神经功能具有一定自愈性。
[Abstract]:Ischemic stroke is the most common type of stroke disease, and its pathogenesis has complex physiological and pathological mechanisms. In recent years, the concept of neurovascular unit (NVU) has opened up a new direction for the study of pathological mechanism and treatment strategy of ischemic stroke. Multi-target therapy for structural damage in NVU will have a better effect on the recovery of neurological function and overall prognosis after stroke. Scalp acupoint acupuncture is a kind of scalp acupuncture method summarized by Professor Yu Zhishun according to the basic theory system and combined with the unique clinical experience in practice. The therapeutic effect of scalp acupuncture on ischemic stroke is definite. According to its "field" hypothesis, Scalp acupuncture has a wide range of regulatory effects on brain tissue. The mechanism of head acupoint cluster acupuncture in treating ischemic stroke may be related to promoting the recovery of neurovascular unit structure and function. Neuron is one of the components of NVU and plays a leading role in it. Synapse is the site of neural signal transduction between neurons or between neurons and effector cells, which plays an important role in the interrelation and influence of each component of NVU. Synaptophysin (synapsin, SYN) is a calcium binding protein, which is specifically distributed on the synaptic vesicle membrane. The increase of SYN expression after ischemic stroke marks the reconstruction of synaptic structure and function and the recovery of neuronal function in NVU. In order to verify the therapeutic mechanism of cephalic plexus acupuncture on ischemic stroke, this study designed an animal experiment on the effect of scalp cluster acupuncture on synaptophysin SYN expression in the penumbra of rats. Aim: to observe the effect of scalp acupuncture on the neurologic function score and synaptophysin SYN positive area in the penumbra of rats with cerebral ischemia, and to study the mechanism of the recovery of nerve function after ischemic stroke by scalp acupuncture. Methods: 54 male SD rats were divided into three groups: group A (group A), group B (model group) and group C (sham operation group), with 18 rats in each group. Each group was randomly divided into 3 subgroups according to 3 time points (1 day / 7 days / 14 days). At each time point, 6 rats in each group were treated with modified thread embolization method. Group C only performed sham-operation. Group A rats were treated with scalp cluster prick method and group B and C were not treated with any intervention. The nerve function was evaluated by Bederson method at 24 hours after model making and 1 hour before the sample was taken, and the SYN expression in the penumbra of the rat brain tissue was photographed by immunofluorescence staining and fluorescence microscope, and the expression of SYN in the penumbra region of the rat brain tissue was photographed. The area percentage of SYN positive expression was calculated by using biological image analysis software. Results 1. The scores of neurological function in group A were significantly lower than those in group B at day 7 and day 14 (P0.05). The scores of neurological function in group A were significantly lower than those in group A at day 7 and day 14 after modeling (P0.05). The neurologic function scores in group B were significantly lower than those in group B at 14 days after modeling (P0.05). The expression of synaptophysin SYN in the penumbra of rats in group A was significantly higher than that in group B on day 7 and day 14 (P0.05). In group A, the expression of SYN in the penumbra was increased gradually at day 1 and day 14 (P0.05). Conclusion: 1. MCAO model can cause neurologic impairment and decrease the expression of synaptophysin SYN in penumbra. The scalp acupoint cluster method can continuously improve the neurological function score of cerebral ischemia rats and has a positive effect on the recovery of neural function. Cephalic plexus needling can significantly increase the expression of synaptophysin SYN in penumbra of rats with cerebral ischemia and promote the synaptic structure and function reconstruction of neurons in NVU. The neural function of rats after cerebral ischemia has a certain self-healing.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245

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