生酮饮食对新生期反复惊厥大鼠远期神经行为及大脑皮层内锌离子流入蛋白7表达的影响

Effects of ketogenic diet on neurobehavioral damages and expression of ZIP7 in cerebral cortex of rat following recurrent neonatal seizures

目的 观察生酮饮食（ketogenic diet,KD）对三氟乙醚所诱导的新生期大鼠反复惊厥后远期神经行为损伤的影响及其大脑皮层内锌离子流入蛋白7（ZIP7）表达的变化.方法 24只8日龄SD大鼠,随机分为4组,分别为空白对照组（NS＋ND组）、对照生酮饮食组（NS＋KD组）、单纯惊厥组（RS＋ND组）、惊厥生酮饮食组（RS＋KD组）,每组n=6.于出生后31d进行前肢悬吊实验和旷场实验.于生后32 d各组大鼠断头取大脑皮层组织,采用免疫印迹技术（Western blot）检测ZIP7的表达.结果 （1）前肢悬吊实验中,RS＋ND组前肢悬吊时间[（19.17±2.48）s]较NS＋ND组[（32.67±2.42）s]明显缩短（P＜0.05）,RS＋KD组前肢悬吊时间[（26.25±2.87）s]较RS＋ND组明显延长（P＜0.05）.（2）旷场实验中,与NS＋ND组[（2.00±0.63）次]相比,RS＋ND组清洁次数[（4.00±0.63）次]明显增多（P＜0.05）;与RS＋ND组比较,RS＋KD组清洁次数[（2.17±0.75）次]明显减少（P＜0.05）.（3） Western blot结果显示,与NS＋ND组相比,RS＋ND组皮层ZIP7表达显著降低（P＜0.05）;与RS＋ND组相比,RS＋KD组皮层ZIP7表达明显升高（P＜0.05）.结论 新生期反复惊厥导致神经行为损伤,生酮饮食可能通过上调大脑皮层中ZIP7的表达改善神经行为。

Objective To explore the intervention effect of ketogenic diet （KD） on neurobehavioral damages after flurothyl-induced neonatal recurrent seizures in rats and on the expression of ZIP7.Methods Postnatal day 8 SD rats （n=24） were divided into four groups randomly：normal group （NS＋ND group）,non-seizure and ketogenic diet group （NS＋KD group）,seizure and normal diet group （RS＋ND group）,seizure and ketogenic diet group （RS＋KD group）,n=6 in eacb group.At postnatal day 31,the grip-strength test and open field test were monitored.At postnatal day 32,rats were sacrificed and the expression of ZIP7 protein level in cerebral cortex was detected with Western blot.Results （1） The grip-strength test：compared with NS＋ND group （（32.67±2.42） s）,the time needed to hold on glass bar in RS＋ND group （（19.17±2.48） s） was shorter significantly （P〈0.05）.Compared with RS＋ND group,the time needed to hold on glass bar in RS＋KD group （（26.25±2.87） s） was significantly longer （P〈0.05）.（2） Open field test：compared with NS＋ ND group （（2.00± 0.63） times）,the times of grooming in RS＋ND group （（4.00±0.63） times） were more （P〈0.05）.Compared with RS＋ND group,the times of grooming in the RS＋KD group （（2.17±0.75） times） were fewer （P〈0.05）.（3）Western blot：compared with NS ＋ND group,the level of ZIP7 of the RS＋ND group in cerebral cortex were lower （P〈0.05）.Compared with RS＋ND group,the level of ZIP7 of the RS＋KD group in cerebral cortex were higher （P〈0.05）.Conclusion Neonatal recurrent seizures may damage neurobehavior,and the neuroprotective effects of ketogenic diet may be associated with the increasing of ZIP7 in cerebral cortex.