左乙拉西坦对硝酸甘油致偏头痛大鼠高颈髓谷氨酸能和GABA能中间神经元的影响

发布时间：2018-04-21 14:55

本文选题：偏头痛 + 左乙拉西坦　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:将SD(Sprague Dawley)大鼠给予左乙拉西坦(LEV)灌胃,观察该药对硝酸甘油所致偏头痛大鼠模型在行为表现方面的影响以及高颈髓谷氨酸能和GABA能中间神经元的变化,探讨LEV在偏头痛方面的防治作用,以及可能的作用机制。方法:1.将24只健康雄性SD大鼠普通喂养一周,采用分层随机法,将其分成假模型组(6只)和模型组(18只)。采取皮下注射硝酸甘油(NTG)的方法,造实验性偏头痛大鼠模型,假模型组大鼠给予皮下生理盐水。造模成功后采用随机方法将模型组中大鼠分为模型对照组(6只)、低剂量干预组(6只)及高剂量干预组(6只)。假模型组:生理盐水灌胃,1次/日,连续1周,待末次灌胃后,进行皮下注入生理盐水。模型对照组:生理盐水灌胃,1次/日,连续1周,待末次灌胃后,进行皮下注入硝酸甘油,再次造模。低剂量干预组:左乙拉西坦100 mg/kg灌胃,1次/日,连续1周,待末次灌胃后,进行皮下注入硝酸甘油,再次造模。高剂量干预组:左乙拉西坦200 mg/kg灌胃,1次/日,连续1周,待末次灌胃后,进行皮下注入硝酸甘油,再次造模。2.造模成功后密切观察每只大鼠在行为学方面的改变。每半个小时作为一个观察时间单元,采取持续时间分段计数的方式观察90min内各个分段时间的大鼠挠头、爬笼和甩头的次数。3.复制偏头痛模型3h后,给予腹腔注射麻醉,进行甲醛灌注,取颈髓1-2。用免疫组化方法对谷氨酸(Glu)和γ-氨基丁酸(GABA)染色,然后用光学显微镜观察谷氨酸和GABA免疫阳性细胞的数目,并在同侧脊髓后角部位随机观察5个高倍视野,依次记录阳性细胞数目。结果:1.行为学变化:与假模型组对照,其余各组大鼠的挠头次数、爬笼次数和甩头次数均呈现明显增加(P0.05)。与模型对照组比较,药物干预组大鼠挠头次数、爬笼次数和甩头次数均随着给药剂量的升高而降低。2.谷氨酸和GABA免疫阳性细胞数目:与假模型组比较,其余各组的谷氨酸免疫阳性细胞数目显著增多(P0.05),GABA免疫阳性细胞数目显著减少(P0.05);与模型对照组比较,低、高剂量干预组大鼠颈髓谷氨酸免疫反应阳性细胞数目明显降低(P0.05),GABA免疫阳性细胞数目明显增多(P0.05),且呈剂量依赖性。结论:左乙拉西坦可以改善偏头痛大鼠在行为学方面的变化,能使高颈髓谷氨酸能中间神经元的数目降低,GABA能中间神经元的数目增高,且呈剂量依赖性,其机制可能与调节高颈髓中间神经元“兴奋-抑制”稳态的平衡有关。
[Abstract]:Aim: to observe the effect of levoxetam (Levo) on the behavior of migraine rats induced by nitroglycerin and the changes of glutaminergic and GABA interneurons in the cervical spinal cord of SD(Sprague Dawley rats. To explore the preventive and therapeutic effect of LEV on migraine and its possible mechanism. Method 1: 1. Twenty-four male Sprague-Dawley (SD) rats were fed for one week and were randomly divided into sham model group (n = 6) and model group (n = 18). The experimental migraine rat model was established by subcutaneous injection of nitroglycerin NTG, and the sham model group was given subcutaneous normal saline. The rats in the model group were randomly divided into control group (n = 6), low dose group (n = 6) and high dose group (n = 6). Sham model group: saline was administered once a day for 1 week, and then subcutaneously injected with normal saline after the last gastric perfusion. The model control group was treated with normal saline once a day for 1 week, then subcutaneously injected nitroglycerin after the last intragastric perfusion, and the model was made again. Low dose intervention group: levoethoxetam 100 mg/kg intragastric perfusion once a day for 1 week, after the last gastric perfusion, subcutaneous injection of nitroglycerin, re-model. High dose intervention group: levoethoxetam 200 mg/kg intragastric perfusion once a day for 1 week, after the last gastric perfusion, subcutaneous injection of nitroglycerin, re-model. 2. The behavioral changes of each rat were closely observed after successful modeling. As an observation time unit every half hour, the number of head scratching, cage crawling and head flick in each segment of 90min was observed by using the method of continuous time piecewise counting. After 3 hours of migraine model, intraperitoneal injection of anesthesia, formaldehyde perfusion, cervical spinal cord 1-2. Glutamate and 纬 -aminobutyric acid (GABA) were stained with immunohistochemical method. The number of glutamate and GABA immunoreactive cells was observed by optical microscope, and five high-power visual fields were observed randomly at the posterior horn of ipsilateral spinal cord. The number of positive cells was recorded in turn. The result is 1: 1. Behavioral changes: compared with the sham model group, the number of head scratching, the number of cage climbing and the number of head flick in the other groups were significantly increased (P 0.05). Compared with the model control group, the number of head scratching, the number of cage climbing and the number of head flick in the drug intervention group decreased with the increase of drug dosage. The number of Glutamic acid and GABA immunoreactive cells: compared with the pseudo-model group, the number of GA-immunoreactive cells in the other groups was significantly increased and the number of GABA-immunoreactive cells in the other groups was significantly decreased than that in the model control group, and was lower than that in the model control group. The number of glutamate immunoreactive cells in the cervical spinal cord decreased significantly in the high dose intervention group, and the number of GABA immunoreactive cells in the cervical spinal cord increased significantly in a dose-dependent manner. Conclusion: levoethylacetam can improve the behavioral changes of migraine rats and decrease the number of glutaminergic intermediate neurons in high cervical spinal cord in a dose-dependent manner. The mechanism may be related to regulating the balance of excitation-inhibition homeostasis of high cervical spinal cord interneuron.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R747.2

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