电针联合丰富康复训练调控MAG和OMgp蛋白对脑缺血神经再生的影响

发布时间：2018-01-16 13:27

本文关键词：电针联合丰富康复训练调控MAG和OMgp蛋白对脑缺血神经再生的影响　出处：《安徽中医药大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:观察电针、丰富康复训练和电针联合丰富康复训练对不同时间点脑缺血再灌注损伤大鼠神经功能和神经再生抑制因子MAG、OMgp蛋白表达的影响。从调控中枢神经再生抑制因素的角度出发,探讨电针联合丰富康复训练对缺血性脑卒中神经功能恢复的作用机制,为针康法治疗缺血性脑卒中提供新的实验依据。方法:1.取SPF级Sprague-Dawley大鼠120只,随机抽取假手术组24只大鼠,其余大鼠采用右侧颈外动脉线栓法制备局灶性大脑中动脉阻塞再灌注模型,随机将造模成功的大鼠分为模型组、电针组(电针治疗)、康复组(丰富康复训练治疗)、针康组(电针联合丰富康复训练治疗),每组24只。各组分为脑缺血后3d、7d、14d三个亚组,一组8只。电针疗法设疏密波5~100次/s,20min/天。丰富康复训练疗法即丰富环境加上平衡木、转棒、网屏、滚筒训练,每项每天10min,丰富环境每3天换一次。2.运用神经功能康复评定法对每组进行神经缺损体征和感觉、运动功能评分;利用HE(苏木素和伊红染色法)染色技术观察脑缺血后大鼠海马CA1区病理形态的变化;应用免疫组织化学技术分析比较各组大鼠海马CA1区MAG、OMgp阳性细胞的表达情况;通过Western Blot检测技术观察MAG、OMgp的蛋白表达变化。结果:1.HE染色光镜下可见假手术组海马CA1区神经细胞染色均匀,形态结构完整,模型组神经细胞水肿明显,排列稀疏,电针组和康复组海马CA1区可见神经细胞肿胀减轻,针康组肿胀的神经细胞明显减少,神经细胞形态结构趋于正常。2.神经功能和感觉、运动功能的评分:在进行电针治疗、平衡木训练、转棒训练、网屏训练后对各组进行评分比较分析发现,假手术组神经功能正常。治疗前,比较模型组和三个治疗组的评分,差异无统计学意义(P0.05)。治疗3d后,三个治疗组的评分与模型组比较,均有降低,但无统计学意义(p0.05),三个治疗组之间比较,差异亦无统计学意义(p0.05);治疗7d、14d后,三个治疗组的评分与模型组比较,呈降低趋势,电针治疗组和丰富康复训练治疗组评分明显降低,差异具有统计学意义(p0.05),电针联合丰富康复训练治疗组评分显著降低,差异具有统计学意义(p0.01)。三个治疗组评分比较,电针治疗组和丰富康复训练治疗组评分比较差异无统计学意义(p0.05),7d时,电针联合丰富康复训练治疗组评分比电针治疗组和丰富康复训练治疗组低,差异有统计学意义(p0.05);14d时,电针联合丰富康复训练治疗组比电针治疗组和丰富康复训练治疗组评分明显降低,差异具有统计学意义(p0.01)。3.mag、omgp蛋白的表达:脑缺血后第3d、7d、14d对大鼠海马ca1区进行免疫组织化学染色和westernblot技术检测,通过统计分析可知,假手术组缺血侧海马ca1区中mag、omgp蛋白呈基础表达,其余四组缺血侧海马ca1区mag、omgp蛋白均有较强程度表达,模型组mag、omgp蛋白在海马ca1区中的表达高于假手术组(p0.01)。术后第3d,四组mag、omgp蛋白表达均呈增强趋势,但与模型组比较,电针治疗组、丰富康复训练治疗组和电针联合丰富康复训练治疗组的蛋白表达均无统计学意义(p0.05);术后第7d,四组mag、omgp蛋白表达达到高峰,第14d有下降趋势,但仍高于术后第3d。术后7d、14d与模型组相比,电针治疗组、丰富康复训练治疗组mag、omgp蛋白表达有所降低(p0.05),针康治疗组表达降低明显,差异有统计学意义(p0.01)。三个治疗组mag、omgp蛋白表达相比较,电针治疗组和丰富康复训练治疗组之间蛋白表达比较,差异无统计学意义(p0.05);7d时,电针联合丰富康复训练治疗组与电针治疗组、丰富康复训练治疗组比较,mag、omgp蛋白表达降低,差异具有统计学意义(p0.05),14d时,电针联合丰富康复训练治疗组与电针治疗组和丰富康复训练治疗组mag、omgp蛋白表达相比,差异具有统计学意义(p0.01)。结论:1.脑缺血损伤后,电针治疗、丰富康复训练和针康疗法均具有改善缺血侧海马病理性损害和促进神经功能缺损恢复的作用,且针康治疗比单纯的电针治疗和康复训练治疗更有效;2.针康法能够抑制神经再生抑制因子MAG、OMgp蛋白的表达,从而促进神经功能的恢复。
[Abstract]:Objective: To observe the effects of electroacupuncture combined with rehabilitation training, rich and rich in rehabilitation training on cerebral ischemia reperfusion injury at different time points in rat nerve function and nerve regeneration inhibitory factor MAG and OMgp protein expression. From the regulation of central nerve regeneration inhibitory factors point of view, to explore the rich mechanism of electro acupuncture combined with rehabilitation training on the recovery of stroke ischemic nerve function, to provide new experimental evidence for acupuncture and rehabilitation therapy on ischemic stroke. Methods: 1. SPF 120 Sprague-Dawley rats, randomly selected 24 rats in the sham operation group, the rest rats using the right external carotid artery occlusion of middle cerebral artery occlusion reperfusion model randomly, model rats were divided into model group, electroacupuncture group (electroacupuncture), rehabilitation group (rich rehabilitation treatment), treatment group (electro acupuncture combined with rehabilitation training in the treatment of the rich), 24 rats in each group. Each component For 3D after cerebral ischemia, 7d, 14d three groups, a group of 8 rats. Electroacupuncture dilatational wave 5~100 /s set, 20min/ days. The rich rich environment rehabilitation training therapy plus the balance beam, rotating rod, roller screen, training, every day 10min, rich environment for every 3 days of nerve defect signs and feeling for each neurological rehabilitation evaluation method of a.2. application, the motor function score; using HE (hematoxylin and eosin staining) staining of pathological changes in hippocampal CA1 region of rats after focal cerebral ischemia; analysis and comparison of MAG in hippocampal CA1 region of rats by using immunohistochemical technique, the expression of OMgp positive cells; Blot through Western detection technique to observe the changes of the expression of OMgp MAG protein. Results: 1.HE staining under light microscope, sham operation group of nerve cells in hippocampal CA1 region of uniform dyeing, structure integrity, model group and edema of nerve cells, sparsate, electroacupuncture Group and rehabilitation group in hippocampus CA1 area visible nerve cell swelling, swelling of the needle Kang group obviously reduced the nerve cells, morphology of nerve cells to normal.2. neural function and sensory, motor function score in electro acupuncture treatment, balance training, rotating bar training, after training on each screen score comparison analysis found that the sham operation group of neural function normal. Before treatment, compared with model group and three treatment groups were no significant difference (P0.05). After 3D treatment, compared with three in the treatment group and model group scores were decreased, but there was no statistical significance (P0.05). The comparison between the three treatment groups, the difference is no statistical significance (P0.05); the treatment of 7D, 14d after the three treatment groups were compared with the model group, EA group decreased, and rich in the rehabilitation group were significantly lower, the difference was statistically significant (P0.05), electro acupuncture combined Hefeng Beverly rehabilitation training treatment group score decreased significantly, the difference was statistically significant (P0.01). Three groups were compared, no significant differences in scores between the treatment group and rich rehabilitation group (P0.05), 7d, electro acupuncture combined with rehabilitation training in the treatment group were rich than treatment group and Enriched Rehabilitative Training the treatment group, the difference was statistically significant (P0.05); 14d, electro acupuncture combined with rehabilitation training in the treatment group than the rich rich treatment group and rehabilitation group were significantly decreased, the difference was statistically significant (P0.01.3.mag), the expression of OMgp protein after cerebral ischemia: 3D, 7d, 14d were detected by immunohistochemistry staining and Westernblot technique on hippocampal CA1, through statistical analysis, the sham operation group of ischemic side in CA1 region of hippocampus MAG, basal expression of OMgp protein is, the other four groups in hippocampus CA1 area of ischemic MAG, OMgp egg White are strong degree of expression of MAG in the model group, the expression of OMgp protein in hippocampal CA1 region in higher than that of sham operation group (P0.01). 3D after operation, four groups of MAG, OMgp protein expression was increased, but compared with the model group, EA group, rich in the rehabilitation group and the electroacupuncture combined with rich the rehabilitation group expression had no statistical significance (P0.05); 7d after operation, four groups of MAG, OMgp protein expression reached the peak, the 14d decreased, but still higher than the 3D. after the first postoperative 7d, 14d compared with the model group, electroacupuncture group, rehabilitation group MAG rich. The expression of OMgp protein decreased (P0.05), acupuncture treatment group expression decreased significantly, the difference was statistically significant (P0.01). The three MAG treatment groups, the expression of OMgp protein in comparison between treatment group and rehabilitation group is rich in protein expression, the difference was not statistically significant (P0.05); 7d, EA With rich rehabilitation training treatment group and electroacupuncture group, rehabilitation group, rich MAG, OMgp protein expression decreased, the difference was statistically significant (P0.05), 14d, electro acupuncture combined with enriched rehabilitation training in the treatment group and EA group and enrich the rehabilitation group MAG, the expression of OMgp protein compared, the difference was statistically significant (P0.01). Conclusion: 1. after cerebral ischemic injury, acupuncture, rehabilitation training and acupuncture therapy rich can improve ischemic hippocampus pathological damage and promote the nerve function recovery effect, and acupuncture treatment is more effective than acupuncture treatment and rehabilitation treatment alone; 2. electro acupuncture can inhibit nerve regeneration inhibitory factor MAG, OMgp protein expression, so as to promote the recovery of nerve function.

【学位授予单位】：安徽中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245

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