新生期反复惊厥大鼠神经行为改变及溶酶体酶抑制剂的干预作用  

Recurrent neonatal seizure induced behavioral changes and the intervention by cathepsin B inhibitor

作　　者：闫建珍[1] 倪宏[1] 张乐玲[1] 

机构地区：[1]苏州大学附属儿童医院儿研所,苏州215003

出　　处：《中华行为医学与脑科学杂志》2011年第3期214-217,共4页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：国家自然科学基金（30870808）;江苏省自然科学基金（BK2007509,BK2010233）;江苏省高校自然科学基金（07KJB320103）

摘　　要：目的探讨溶酶体酶抑制剂（CBI）在三氟乙醚致新生期大鼠反复惊厥性脑损伤中的干预作用及其信号通路。方法90只生后6d（P6）的健康SD大鼠随机分为3组（n=30）：对照（CONT）组、反复惊厥（RS）组及CBI组，RS组每日吸入三氟乙醚诱导惊厥发作5次，每次间隔30min，连续9d；CONT组同样操作但不吸入三氟乙醚；CBI组惊厥前腹腔注射CBI（总量2μl），同样方法吸入三氟乙醚诱导惊厥，方法同RS组。分别于生后不同时间段检测体格、神经发育以及认知情感能力，并于末次惊厥后1．5h、3h、6h、24h及第35天五个时间点（n=6）取大脑皮层组织，采用免疫印迹技术检测自噬相关蛋白cathepsin B的表达。结果P14时Rs组体质量（27．28±1．60）k，低于CONT组（33．45±1．57）kg，差异具有显著性（P〈0．0001）。神经发育指标显示悬崖回避时间，日龄P12时RS组历时（2．10±1．45）s，延长于CBI组（1．05±0．32）s，差异有显著性（P〈0．05）。旷场行为测试：RS组水平运动（20．00±13．08）次，少于CONT组（57．83±33．22）次，差异有显著性护〈0．05）；RS组垂直运动（2．50±2．43）次，少于CONT组和CBI组[分别为（22．17±10．74）次，（9．33±5．39）次]，差异有显著性（P〈0．01）。Western blot法检测，RS组在1．5h、3h、6h、24h各时间点CathepsinB的表达明显高于CONT组（P〈0．05）；CBI组于1．5h、3h、6h、24h各时间点cathepsin B的表达明显低于RS组妒〈0．05）。P35时间点3组之间差异无显著性（P〉0．05）。结论CBI能够通过自噬信号途径抑制发育期惊厥所致神经行为损伤。Objective To explore the intervention effect of cathepsin B inhibitor （CBI） and its signaling pathways after flurothyl inducing brain injury in rats with recurrent seizures. Methods 6-day-old （P6） SD rats were randomly divided into： recurrent neonatal seizure group （RS group, n = 30）, CBI-treated seizure group （CBI group, n = 30） and the control group（ CONT group）. Rats in RS group were subjected to 5 seizures with flurothyl during the first 14 days of life. In CBI group, CBI was injected each day before seizures were induced. Then examined development indexes such as the physical growth, neural reflex, neural behavior and cognitive emotional com- petence. Western blot was employed to determine Cathepsin B expression at different time points （1.5h,3h,6h, 24h and P35） after the last convulsion. Results The weights of rats in RS group（ （27.28 ± 1.6） kg） were light- er than CONT group（ （33.45 ± 1.57） kg）. They had significant difference （P〈0.01） at the age of p14. The time of cliff avoidance in RS group（ （2.10 ± 1.45）s） was longer than CBI group（ （ 1.05 ±0.32）s）. It showed a statistically significant （P 〈 0.05） in pl 2. In the open-field behavior test ： the activities of RS group （20.00 ± 13.08 ） were markedly reduced than CONT group （57.83 ± 33.22） in the horizontal movements, the RS group （2.50 ± 2.43） were significantly decreased than the CONT group and CBI group（ （22.17 ± 10.74）, （9.33 ±5.39） ） in the vertical motions （P 〈 0.05 ）. Cathepsin-B expression in RS group （ 1.5h,3h, 6h and 24h）was significantly higher than that at the same time in CONT group（P〈 0.05 ）. Cathepsin-B expression of CBI group was signifieantly decreased compared with that in RS group （P〈 0.05 ） at the time point （ 1.5h ,3h,6h and 24h）. There were no significant differences among three groups at P35 （P 〉 0.05 ）. Conclusions CBI can improve brain injury and regulate the expression of abnormal molecules.

关 键 词：惊厥 新生期 生长发育 旷场实验 溶酶体酶抑制剂 

分 类 号：R722.1[医药卫生—儿科]