摘要
目的观察内侧颞叶癫癎(mesial temporal lobe epilepsy,MTLE)患儿及MTLE大鼠海马组织中dy-namin-1和磷酸化dynamin-1蛋白的表达变化,探讨其在MTLE发生发展中的作用。方法 25日龄Sprague-Dawley雄性大鼠随机分为急性期对照组、急性期模型组、潜伏期对照组、潜伏期模型组、慢性期对照组、慢性期模型组,应用氯化锂-匹罗卡品诱导建立MTLE动物模型。同时收集接受神经外科手术、并经病理结果确认存在海马硬化的5例患儿的海马组织,及病理科尸检4例对照组海马组织。提取海马组织蛋白,采用Western blot方法和免疫组织化学技术检测总dynamin-1蛋白及磷酸化dynamin-1蛋白在MTLE患儿及不同时间点MTLE大鼠海马组织中的表达变化。结果 Western blot显示磷酸化dynamin-1蛋白在MTLE大鼠急性期及慢性期表达较同时期对照组显著降低(P<0.05);同时,磷酸化dynamin-1蛋白在MTLE患儿海马组织表达较对照组显著下降(P<0.05)。而dy-namin-1总蛋白在MTLE大鼠各期、MTLE患儿及对照组海马之间表达无明显差异(P>0.05)。免疫组化结果显示磷酸化dynamin-1在对照组大鼠急性期、慢性期、对照组患儿海马神经元胞浆中高表达,而在同时期MTLE大鼠和MTLE患儿海马神经元胞浆中表达下降(P<0.05)。结论磷酸化dynamin-1而不是dynamin-1总蛋白在MTLE患儿及MTLE大鼠出现癎性发作期间表达下调,其表达变化提示dynamin-1可能通过磷化酸/去磷酸化方式参与MTLE的发生发展。
Objective To observe the expression of dynamin-1 and phospho-dynamin-1 in the hippocampus of children and rats with mesial temporal lobe epilepsy （ MTLE ） and to investigate the roles of dynamin-1 and phosphodynamin-1 in the development of MTLE. Methods Male Sprague-Dawley rats （ aged 25 days） were randomly divided into acute control （AC）, acute seizure （AS）, latent control （LC）, latent seizure （LS）, chronic control （CC） and chronic spontaneous seizure （CS） groups. Lithium chloride-pilocarpine was used to induce a rat model of MTLE. The hippocampus samples of 5 children with a pathologically confirmed hippocampal sclerosis who received surgical operation were collected as a human model （ HM ） group, and the hippocampus samples of 4 dead children （ without organic lesion of the hippocampus） were collected by autopsy as a human control （HC） group. The expression of dynamin-1 and phosphodynamin-1 in the hippocampus of children and rats with MTLE was measured by Western blot and immunohistochemistry. Results The Western blot showed that the expression of phospho-dynamin-1 was significantly lower in the AS and CS groups than in the corresponding control groups （AC and CC groups） （P 〈 0.05 ）. The expression of phospho-dynamin-1 was significantly lower in the HM group than in the HC group （ P 〈 0.05 ）. There were no significant differences in the expression of dynamin-1 among the AS, LS and CS groups and between the HM and HC groups （ P 〉 0. 05 ）. The immunohistochemical results showed that phospho-dynamin-1 was highly expressed in the cytoplasm of hippocampal neurons of AC, CC and HC groups, but its expression was significantly reduced in the AS, CS and HM groups （ P 〈 0.05 ）. Conclusions The expression of phospho-dynamin-1, not dynamin-1, is downregnlated in the hippocampus of children and rats with MTLE during seizures, which suggests that the phosphorylation/dephosphorylation of dynamin-1 may be involved in the development of MTLE.
出处
《中国当代儿科杂志》
CAS
CSCD
北大核心
2013年第2期133-137,共5页
Chinese Journal of Contemporary Pediatrics
基金
国家自然科学基金(30901631)
高等学校博士学科点专项科研基金(200901621 10041)
