EGFR参与电针防治脑缺血的机制及电针治疗缺血性脑卒中的临床观察

发布时间：2018-05-05 15:31

本文选题：电针 + 表皮生长因子受体　；参考：《湖北中医药大学》2017年硕士论文

【摘要】：目的:本课题包括实验研究和临床观察两个方面。临床部分通过对比电针与常规针刺治疗脑缺血的疗效差异,优化脑缺血的临床治疗手段。动物实验部分通过建立小鼠脑缺血模型,探究表皮生长因子受体(Epidermal Growth Factor Receptor,EGFR)在脑缺血再灌注损伤中的功能,以及电针治疗缺血性脑卒中的机制与其的关联性。方法:1.实验研究:将30只C57小鼠以随机数字查表原则,分为对照组、模型组及电针治疗组三个组别,每组各10只小鼠,以线栓法建立小鼠大脑中动脉栓塞(Middle cerebral artery occlusion,MCAO)脑缺血模型,电针组于造模前及MCAO模型完成后分别予以电针刺激“水沟”、“承浆”二穴。各组实验小鼠在缺血再灌注24小时后断头、取脑,一部分直接冷冻做TTC染色,观察小鼠大脑脑缺血体积的变化情况。另一部分分离大脑皮质,提取蛋白,以免疫印迹方法,检测EGFR及i NOS在小鼠大脑皮质的表达变化。2.临床观察:本课题研究设计一个单盲、随机、对照性试验来观察缺血性脑损伤的电针疗效情况,收集武汉市中西医结合医院符合纳入标准的病例,共收集缺血性脑卒中急性期患者60例,采用随机对照分组,分为治疗组和对照组,每组各随机分配30例病例。治疗组予以电针治疗,对照组予以普通针刺治疗,两组患者每天予以电针治疗1次,连续7天为一个疗程,然后间隔一天后进行再下一个疗程,共治疗2个疗程。所有病例均于治疗前后分别采用美国国立卫生院卒中量表(NIHSS评分)(见附录四)、改良的Barthel指数(BI)(见附录三)进行评估,记录治疗前后两组间评分的变化,并将所得结果进行统计学分析,经统计学分析后确定其是否具有统计学意义。基础治疗:参照《中国急性缺血性脑卒中的诊治指南2010》所提及的治疗方案。主要包括改善循环,抗血小板凝集,控制血压,调节血糖,降脂稳定血管斑块,保护及营养神经,对症治疗,防治并发症的常规药物治疗等。结果:1.动物实验结果1.1Bederson评分结果对照组Bederson评分为0分,小鼠表现为无运动功能障碍;模型组进行MCAO手术后,Bederson评分明显上升,与对照组有显著差异,且差异具有统计学意义(p0.05);电针组电针干预后Bederson评分明显低于模型组,且差异具有统计学意义(p0.05)。1.2 TTC染色结果模型组小鼠脑缺血体积要显著高于对照组,表明小鼠进行MCAO手术后,造成了小鼠大脑中动脉供血区的梗塞而引起了小鼠大脑缺血损伤,显示小鼠脑缺血主要部位集中在顶叶,额叶及基底节区等区域;相比较于模型组的结果,电针组显著缩小了小鼠大脑脑缺血的体积。1.3 EGFR及i NOS表达情况相对于对照组而言,模型组小鼠EGFR表达有所上升,差异具有统计学意义(P0.05);电针组电针介入后EGFR蛋白含量较模型组相比继续上升,差异具有统计学意义(P0.05);后各组i NOS表达情况:模型组小鼠i NOS表达含量显著上升,差异具有统计学意义(P0.05);电针介入后i NOS表达含量较模型组相比有所下降,差异具有统计学意义(P0.05);2.临床观察结果2.1治疗后两组NIHSS评分较治疗前均有所降低,与治疗前相比,差异具有统计学意义(均PO.O5);治疗后两组NIHSS评分相比,差异具有统计学意义(PO.05)。2.2治疗两组BI较治疗前均有所上升,与治疗前相比,差异具有统计学意义(均PO.O5);治疗后两组BI相比,差异具有统计学意义(PO.05)。2.3治疗后,治疗组临床总有效率为93.3%,对照组临床总有效率为83.3%。两组有效率经过对比,治疗组疗效明显优于对照组,差异具有统计学意义(PO.05)。结论1.电针可以降低小鼠脑缺血后Bederson评分,减小脑缺血体积,改善小鼠运动功能障碍及神经功能缺损,有助于小鼠的神经功能恢复,其在脑缺血后的神经保护功能的机制可能是电针通过改变星形胶质细胞的分泌表型,使内源性EGFR蛋白合成并且分泌得到促进,并能够抑制i NOS的表达,减少NO的产生,控制中枢神经系统发生的炎性反应,保护脑神经元。2.在予以临床常规用药的基础上,相较于常规针刺而言,结合电针刺激治疗,对缺血性中风患者有更好的效果,缓解急性缺血性中风患者的神经系统的功能缺损程度及症状,改善患者的日常生活能力,可以下降缺血性中风的致残率、致死率。
[Abstract]:Objective: this topic includes two aspects of experimental research and clinical observation. In the clinical part, the clinical therapeutic means of cerebral ischemia are optimized by comparing the therapeutic effects of electroacupuncture and conventional acupuncture in the treatment of cerebral ischemia. In the animal experiment, the Epidermal Growth Factor Receptor (EGFR) is explored by establishing the model of cerebral ischemia in mice. The function of cerebral ischemia reperfusion injury and the mechanism of electroacupuncture in the treatment of ischemic stroke. Methods: 1. experimental study: 30 C57 mice were divided into the control group, the model group and the electroacupuncture treatment group were divided into the control group, the model group and the electroacupuncture treatment group, 10 rats in each group, and the middle cerebral artery embolism (Middle C) was established by the thread emboli method. Erebral artery occlusion, MCAO) cerebral ischemia model, electroacupuncture group before and after the completion of MCAO model to stimulate the "water ditch", "pulp" two points. Each group of experimental mice after 24 hours of ischemia-reperfusion, brain, a part of the direct freezing of TTC staining, observe the cerebral ischemic volume changes in the brain of mice. Separate the cerebral cortex, extract the protein, and detect the expression of EGFR and I NOS in the cerebral cortex of mice by immunoblotting,.2. clinical observation: this subject is designed to design a single blind, random, controlled trial to observe the effect of Electroacupuncture on ischemic brain injury, and collect the cases of the integrated traditional Chinese and Western medicine hospital in Wuhan. 60 cases of acute ischemic stroke were collected and divided into treatment group and control group randomly, divided into treatment group and control group, each group was randomly assigned 30 cases. The treatment group was treated with electroacupuncture, the control group was treated with common acupuncture, the two groups were treated with electroacupuncture 1 times a day for 7 days, and then another day after the next one. A total of 2 courses were treated. All cases were treated with the National Institutes of Health Stroke Scale (NIHSS score) before and after treatment (see Appendix four), the improved Barthel index (BI) (see Appendix three), and recorded the changes of the scores between the two groups before and after the treatment, and the results were statistically analyzed and determined by statistical analysis. No statistical significance. Basic treatment: reference to the treatment guidelines referred to in the guidelines for the diagnosis and treatment of acute ischemic stroke in China 2010>, mainly including improvement of circulation, antiplatelet aggregation, control of blood pressure, regulating blood sugar, reducing fat and stabilizing vascular plaques, protecting and nourishment nerve, treating disease, preventing complications by conventional medication, and so on. Results 1. The result of 1.1Bederson score was 0 in the control group and in the control group, the Bederson score was 0, and the mice showed no motor dysfunction. After the MCAO operation in the model group, the Bederson score increased significantly, and the difference had significant difference with the control group, and the difference was statistically significant (P0.05). The Bederson score in the electroacupuncture group was significantly lower than that in the model group, and the difference was poor. The volume of cerebral ischemia in the model group of P0.05.1.2 TTC staining model group was significantly higher than that of the control group. It showed that after MCAO operation, the infarct of the middle cerebral artery blood supply area in mice caused the cerebral ischemia injury in mice, which showed that the main parts of cerebral ischemia in the mice were concentrated in the parietal lobe, the frontal lobe and the basal ganglia region. Compared with the model group, the expression of.1.3 EGFR and I NOS in the cerebral ischemia of mice was significantly reduced by the electroacupuncture group compared with the control group, the EGFR expression of the model mice increased, the difference was statistically significant (P0.05). The content of EGFR protein in the electroacupuncture group was higher than that in the model group after the electroacupuncture group, and the difference had the difference. Statistical significance (P0.05); the expression of I NOS in each group: the expression of I NOS in the model group was significantly increased, the difference was statistically significant (P0.05); the I NOS expression content decreased compared with the model group after the electroacupuncture intervention, and the difference was statistically significant (P0.05); 2. the two groups of clinical observation results were compared to those of the two groups after 2.1 treatment. Compared with before treatment, the difference was statistically significant (all PO.O5), and the difference between the two groups of NIHSS scores after treatment was statistically significant (PO.05) the two groups of BI were higher than before the treatment, and the difference was statistically significant compared with that before the treatment (PO.O5); the difference was statistically significant (PO.05).2.3 treatment compared to the two groups of BI after treatment. After treatment, the total effective rate of the treatment group was 93.3%, the total effective rate of the control group was 83.3%. two, the curative effect was compared, the curative effect of the treatment group was obviously superior to the control group, the difference was statistically significant (PO.05). Conclusion 1. electroacupuncture can reduce the Bederson score of the mice after cerebral ischemia, reduce the volume of cerebellar ischemia, and improve the motor dysfunction and nerve in mice. Functional defect can help the recovery of neural function in mice. The mechanism of neuroprotective function after cerebral ischemia may be that electroacupuncture can promote the synthesis and secretion of endogenous EGFR protein by changing the secretory phenotype of astrocytes, and can inhibit the expression of I NOS, reduce the production of NO, and control the inflammation of the central nervous system. On the basis of clinical routine medication, the protection of brain neuron.2. is better than conventional acupuncture combined with electroacupuncture. It has a better effect on patients with ischemic stroke, relieving the degree of function defect and symptoms of the nervous system in patients with acute ischemic stroke, improving the daily living ability of the patients, and reducing the ischemic stroke. The rate of disability and death rate of stroke.

【学位授予单位】：湖北中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R246.6

【参考文献】