摘要
目的探讨发育期幼鼠反复惊厥后脑内丛生蛋白的表达及溶酶体酶抑制剂E-64 d对其表达的干预作用。方法日龄21 d的SD大鼠随机分为3组:惊厥组(RS组,n=24)、E-64 d干预组(ERS组,n=24)和对照组(CON组,n=19)。于大鼠21日龄开始,RS组大鼠隔日腹腔注射青霉素(5.1×106U.kg-1.d-1),连续6次诱导惊厥发作;ERS组于每次惊厥前腹腔注射4μg E-64 d,采用同样方法腹腔注射青霉素诱导惊厥发作;CON组大鼠予相应剂量的9 g.L-1盐水腹腔注射。于末次惊厥后21 d(出生后51 d),依据Racine标准从RS组及ERS组各随机选取点燃成功的6只大鼠作为本实验的RS组和ERS组。从CON组随机选取6只作为本实验的CON组。处死18只大鼠,并取其大脑海马及皮质,采用免疫印迹技术(Western blot)检测18只大鼠大脑海马、皮质中丛生蛋白的表达。各组蛋白水平应用SPSS 17.0软件进行统计学分析。结果 RS组大鼠海马及皮质丛生蛋白表达水平均较CON组显著增高,差异均有统计学意义(Pa<0.05)。ERS组海马丛生蛋白的表达低于RS组,差异有统计学意义(P<0.05);ERS组皮质丛生蛋白的表达与RS组比较差异无统计学意义(P>0.05)。结论丛生蛋白参与惊厥性脑损伤的过程,E-64 d的作用可能与其下调丛生蛋白的表达有关。
Objective To explore the expression of Clusterin-α in brain and the intervention effect of lysosomal enzyme inhibitor E-64 d on brain damage of developmental rats with recurrent seizures. Methods Sprague-Dawley（SD） rats at the age of 21 days were randomly divided into recurrent prolonged seizure group（RS group,n=24）,E-64 d-treated seizure group（ERS group,n=24）,normal saline control group（CON group,n=19）.At postnatal day 21st,the penicillin（5.1×106 U·kg-1·d-1） was used to induce seizure attack,6 times.Recurrent seizures were induced every other day in 6 consecutive days in the RS group and ERS group.In ERS group,E-64d（4 μg） was injected intraperitoneally every other day before seizure induced.Rats in CON group were injected with equal amount of normal sodium at the same time. At 21d after last time induced seizure（postnatal days 51）,selected randomly each 6 rats that up to the Racine standard from RS group and ERS group as RS and ERS group in the experiment.At the same time,selected randomly 6 rats from the CON group as the CON group in the experiment.The 18 rats had been slaughtered to take the hippocampus and cerebral cortex at 51-day-old.Clusterin-α levels in hippocampus and cerebral cortex were detected by western blot method. All data were analyzed by SPSS 17.0 software. Results The levels of Clusterin-α in hippocampus and cerebral cortex of RS group were increased significantly compared with that of CON group（Pa〈0.05）.The level of Clusterin-α of ERS group in hippocampus was decreased compared with that of RS group（P〈0.05）.There were no significant difference in Clusterin-α expressions between ERS and RS group（P〉0.05）. Conclusions Clusterin-α may be involved in the pathophysiology of the brain damage resulting from recurrent seizure.E-64 d protects the brain by down-regulating the expression of Clusterin-α.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2012年第12期904-906,共3页
Journal of Applied Clinical Pediatrics
基金
国家自然科学基金(30870808)
江苏省自然科学基金(BK2007509
BK2010233)
江苏省卫生厅医学重点人才项目(RC2011113)
关键词
惊厥
发育期
丛生蛋白
E-64d
大鼠
seizure； developmental period； Clusterin-α； E-64 d； rat
