CAJ | 학술논문

目的探讨突触外GABAA受体的α5亚基在大鼠岛叶电刺激癫痫模型海马中的表达及意义.方法将96只雄性SD大鼠按随机数字表法分为模型组、假手术组和对照组(n=32),每组按时间点分为两个亚组(n=16).模型组:通过慢性电刺激大鼠岛叶建立点燃模型；假手术组:只植入电极而不进行电刺激；对照组:不给予任何处理.各组按时间点断头取脑,应用尼氏染色检测大鼠海马神经元的情况；应用免疫组织化学对α5亚基相关蛋白进行检测；应用实时荧光定量聚合酶链式反应对海马α5亚基mRNA进行检测.结果尼氏染色检测表明癫痫大鼠与正常大鼠的海马各区神经元数目的差异无统计学意义(P＞0.05).免疫组织化学检测显示对照组、假手术组、癫痫组1d时相关蛋白的表达分别为0.13±0.03、0.15±0.06、0.09±0.02,在7d时分别为0.14±0.03、0.13±0.05、0.06±0.01,实时荧光定量PCR检测发现对照组、假手术组、癫痫组相对表达倍数1d时分别为2.67±0.18、2.52±0.21、0.81±0.12,在7d时分别为2.80±0.19、2.69±0.17、0.47±0.11.通过分析发现点燃成功1d时海马α5亚基mRNA及相关蛋白均表现出明显的下降(P＜0.01),并表现出持续的下降,在点燃7d时已经下降到非常低的水平.结论大鼠岛叶电刺激癫痫模型海马中含α5亚基的突触外GABAA受体表达显著减少,含α5亚基的突触外GABAA受体与岛叶癫痫的发生发作有着密切的联系.
목적 탐토돌촉외GABAA수체적α5아기재대서도협전자격전간모형해마중적표체급의의.방법 장96지웅성SD대서안수궤수자표법분위모형조、가수술조화대조조(n=32),매조안시간점분위량개아조(n=16).모형조:통과만성전자격대서도협건립점연모형；가수술조:지식입전겁이불진행전자격；대조조:불급여임하처리.각조안시간점단두취뇌,응용니씨염색검측대서해마신경원적정황；응용면역조직화학대α5아기상관단백진행검측；응용실시형광정량취합매련식반응대해마α5아기mRNA진행검측.결과 니씨염색검측표명전간대서여정상대서적해마각구신경원수목적차이무통계학의의(P＞0.05).면역조직화학검측현시대조조、가수술조、전간조1d시상관단백적표체분별위0.13±0.03、0.15±0.06、0.09±0.02,재7d시분별위0.14±0.03、0.13±0.05、0.06±0.01,실시형광정량PCR검측발현대조조、가수술조、전간조상대표체배수1d시분별위2.67±0.18、2.52±0.21、0.81±0.12,재7d시분별위2.80±0.19、2.69±0.17、0.47±0.11.통과분석발현점연성공1d시해마α5아기mRNA급상관단백균표현출명현적하강(P＜0.01),병표현출지속적하강,재점연7d시이경하강도비상저적수평.결론 대서도협전자격전간모형해마중함α5아기적돌촉외GABAA수체표체현저감소,함α5아기적돌촉외GABAA수체여도협전간적발생발작유착밀절적련계.
Objective To explore the expression of α5 subunit of extrasynaptic GABAA receptor in insular electrical kindled rats and analyze its significance.Methods A total of 96 male Sprague-Dawley rats were divided randomly into kindled,sham-operated and control groups(n =32).And each group was further divided into 2 sub-groups at different time points.Kindled group:insular electrical kindled via chronic electrical stimulation; Sham-operated group:electrode implantation without electrical stimulation;Control group:no operation at all.The number of hippocampal neurons was detected by Nissl staining; the mRNA of α5 subunit of hippocampus by quantitative-polymerase chain reaction(q-PCR); the expression of α5 subunit of hippocampus by immunohistochemistry.Results No significant difference existed in the number of hippocampal neurons between epileptic and normal rats(P＞0.05).The mRNA levels of α5 subunit of hippocampus in the sham-operated and control groups were much higher than that of the kindled group at Day 1 and 7 post-sacrificing(P＜0.01).And the hippocampal expressions of α5 subunit were much more in the sham-operated and control groups than that of the kindled group at Day 1 and 7 post sacrificing(P＜0.01).Conclusion The hippocampal expression of α5 subunit decreases markedly in insular epilepsy.And α5 subunit may play an important role in the occurrence and development of insular epilepsy.