蛛网膜下腔注射布比卡因对大鼠脊髓GABA-AR alpha1亚基的影响

发布时间：2019-04-13 11:28

【摘要】：目的 观察蛛网膜下腔注射布比卡因对大鼠脊髓腰膨大γ-氨基丁酸A受体alphal(GABA-ARα1)亚基表达的影响,探讨布比卡因椎管内注射产生镇静作用的机制。方法雄性SD大鼠32只,体重260±20g,随机分为4组:空白对照组(C组),盐水对照组(NC组),0.5%布比卡因10min组(B10组),0.5%布比卡因30min组(B30组),每组8只。C组不做任何处理,取材同下组;其余三组建立大鼠蛛网膜下腔置管模型后,注射生理盐水或0.5%布比卡因20ul,于相应时点断颈椎处死取脊髓腰膨大段,分别进行尼氏染色、免疫荧光染色,观察脊髓神经细胞形态、GABA-ARα1蛋白在脊髓的分布及检测脊髓Ⅱ板层平均光密度的变化;用western-blot检测GABA-ARα1亚基蛋白在总蛋白、细胞浆蛋白及细胞膜蛋白中表达的变化。结果 1.尼氏染色结果,镜下可见各组神经元细胞排列整齐,未见神经元形态改变;2.免疫荧光结果显示,GABA-ARα1蛋白主要分布于脊髓灰质各板层,高倍镜下可见细胞浆及细胞膜上均有该蛋白存在,以Ⅱ板层分布最为密集,各组之间脊髓Ⅱ板层GABA-ARα1平均光密度值无显著改变(P0.05);3.western-blot检测结果显示,各组GABA-ARα1亚基在总蛋白中相对表达量无显著差异(P0.05);各组细胞浆蛋白中GABA-ARα1亚基表达量:B10组、B30组与C组及NC组相比均显著下降(p0.01),且B10组下降更明显;各组细胞膜蛋白中GABAARα1亚基表达量:B10组、B30组、与C组及NC组相比均显著上调(p0.01),且B10组上调更明显;结论 1.蛛网膜下腔注射0.5%布比卡因使GABA-ARα1亚基蛋白从细胞浆转运至细胞膜,上调细胞膜该受体的表达数量,这可能是椎管阻滞引起镇静作用的机制之一。2.蛛网膜下腔注射0.5%布比卡因在作用10min时对GABA-ARα1亚基蛋白细胞膜表达的影响最大。
[Abstract]:Aim to observe the effect of bupivacaine injection into subarachnoid space on the expression of 纬-aminobutyric acid A receptor alphal (GABA-AR 伪 1) subunit in spinal cord of rats, and to explore the mechanism of sedative effect induced by bupivacaine intraspinal injection. Methods 32 male SD rats, weighing 260 卤20g, were randomly divided into 4 groups: blank control group (C group), saline control group (NC group), 0.5% bupivacaine 10min group (B10 group) and 0.5% bupivacaine 30min group (B30 group). There were 8 rats in each group. Group C did not do any treatment, and the material was taken from the same group as the lower group. The other three groups were injected with normal saline or 0.5% bupivacaine 20ul.After establishing the subarachnoid catheterization model in the other three groups, the spinal cord lumbar enlargement segments were killed at the corresponding time point and stained by Nissl's staining and immunofluorescence staining, respectively. The morphology of spinal nerve cells, the distribution of GABA-AR 伪 1 protein in spinal cord and the change of average optical density in lamina 鈪，

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