新生期大鼠反复惊厥后皮质丛生蛋白的表达及生酮饮食的干预作用  

Expression of cortex clusterin and intervention effect of ketogenic diet on neonatal rats with recurrent seizures

作　　者：田甜[1] 孙奇[1] 赵东敬[1] 徐丹凤[1] 倪宏 

机构地区：[1]苏州大学附属儿童医院神经内科,215003 [2]儿科医学研究所

出　　处：《中华实用儿科临床杂志》2014年第9期694-697,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（81271458）;江苏省医学重点人才基金（RC2011113）

摘　　要：目的探讨新生期大鼠反复惊厥后大脑皮质丛生蛋白（CLU）的表达以及生酮饮食对表达的干预作用。方法日龄8d（P8，下同）的SD大鼠共36只，依据随机数字表法随机分为正常对照组、惊厥组和生酮饮食组，每组各12只。于P9，惊厥组及生酮饮食组大鼠吸入三氟乙醚诱导惊厥发作，反复诱导惊厥持续30min，1次／d，同样方法连续诱导8d；正常对照组同样操作但不吸人三氟乙醚。于P35进行神经行为学评分。于P58每组各随机取6只大鼠共18只，断头取大脑皮质，采用免疫印迹技术（Westernblot）检测各组大鼠大脑皮质CLU的表达。结果神经发育指标结果显示：平面翻正实验中，正常对照组、生酮饮食组平面翻正时间[（1．03±0．54）s，（0．89±0．16）s]号惊厥组[（0．64±0．30）S]比较差异均有统计学意义（P均〈0．05）；负向趋地反应实验中，正常对照组、生酮饮食组负向趋地反应时间[（1．92±0．90）s，（5．17±0．72）s]与惊厥组[（7．33±0．65）s]比较差异均有统计学意义（P均〈0．01）；悬崖回避实验中，正常对照组、生酮饮食组悬崖回避时间[（4．33±2．54）s，（8．75±2．26）s]与惊厥组[（16．58±4．25）s]比较差异均有统计学意义（P均〈0．01）。Westernblot结果显示：惊厥组CLU在大脑皮质的表达（2．24±0．53）与正常对照组（1．44±0．11）及生酮饮食组（1．56±0．24）比较差异均有统计学意义（P〈0．01，P〈0．05）。结论新生大鼠反复惊厥远期大脑皮质CLU水平明显上调，提示其可能参与了惊厥性脑损伤的病理生理机制；而生酮饮食可能通过下调CLU水平参与了惊厥性脑损伤的修复过程。Objective To investigate dynamic expressions of cortex clusterin （CLU） and intervention effect of ketogenic diet （KD） on neonatal rats with recurrent seizures. Methods Thirty-six-8-day postnatal SD rats were ran- domly divided into 3 groups ：normal control group（ NS ＋ ND group,n = 12） , and the recurrent-seizure and normal diet group（ RS ＋ ND group, n = 12）, and the recurrent-seizure and KD group（ RS ＋ KD group, n = 12）. From 9 d, rats in RS ＋ ND group and RS ＋ KD group were subjected to recurrent seizures induced by volatile flurothyl 30 min each day for consecutive 8 days. Rats in NS ＋ ND group were placed into the container for an equal amount of time to their coun- terpart without exposure to flurothyl. Scores on neurological behaviors at 35 days postnatally were examined. CLU protein levels in cerebral cortex were determined by Western blot at 58 days postnatally. Results Neurodevelopmental indica- tors analysis ： in the plane righting experiment, there were significant differences between NS ＋ ND group [ （ 1.03 ± 0. 54） s] ,RS ＋ KD group[ （0.89 ±0.16） s] and RS ＋ ND group [ （0.64 ±0.30） s] about the time of plane righting （ all P 〈 0.05 ） ; in the negative geotaxis reaction experiment, the rats of NS ＋ ND group [ （ 1.92 ± 0.90 ） s ] , and RS ＋ KD group [ （5.17 ±0.72） s] about the time of negative geotaxis reaction were significantly different compared with RS ＋ ND group[ （7.33 ±0.65） s] （all P 〈0. 01 ）. In the cliff avoidance test, there were significant differences be- tween NS ＋ ND group, RS ＋ KD group [ （4.33 ± 2.54） s, （ 8.75 ± 2.26 ） s ] and RS ＋ ND group [ （ 16.58 ± 4.25 ） s 1 about the time of cliff avoidance （ all P 〈0.01 ）. Western blot showed that the expression of CLU in cerebral cortex of the RS ＋ ND group [ （ 2.24 ± 0.53 ） s ] was obviously increased compared with NS ＋ ND group [ （ 1.44 ±0.11 ） s ] （P 〈0.01 ） ,and there also had significant difference between

关 键 词：惊厥 新生大鼠 丛生蛋白 生酮饮食 

分 类 号：R742.1[医药卫生—神经病学与精神病学]