生酮饮食对新生期反复惊厥大鼠远期神经行为损伤及相关分子表达的影响
发布时间：2018-09-13 06:38
【摘要】：目的探讨生酮饮食对三氟乙醚诱导的新生期反复惊厥大鼠远期神经行为损伤及相关分子表达的影响。方法日龄8天(用P8表示,下同)的Sprague-Dawley(SD)大鼠24只,按照随机数字表法分为对照组(NS组)和惊厥组(RS组),每组各12只。RS组大鼠吸入三氟乙醚诱导惊厥发作,反复诱导惊厥持续30分钟,每日一次,相同方法连续诱导8天;NS组除不吸入三氟乙醚其余操作相同。于P21依据是否给予生酮饮食(KD)干预,每组大鼠按照随机数字表法再分为两组,即未惊厥普通饮食组(NS+ND组)、未惊厥生酮饮食组(NS+KD组)、惊厥普通饮食组(RS+ND组)、惊厥生酮饮食组(RS+KD组),每组各6只。NS+ND组和RS+ND组给予普通饮食,NS+KD组和RS+KD组给予生酮饮食,饮食干预10天。于KD实施当天、7天、10天每组大鼠尾静脉采血监测血糖及血酮值。自P9起隔天测量并记录每组大鼠的体重。于P27、P31分别进行两次神经行为学测试,包括平面翻正、悬崖回避、前肢悬吊、负向趋地反应实验。P31行旷场实验。于P32大鼠断头取海马及大脑皮质,采用免疫印迹法(western blot)检测各组大鼠海马及大脑皮质中Cathepsin E、ZIP7、MT3、CLU蛋白的表达。结果1、对体重的影响:生酮饮食干预前,RS组P17~P20各时间点体重较NS组对应时间点明显降低(P0.05)。生酮饮食干预期间(P21~P31),四组大鼠的体重变化存在总体性差异(P0.05),进一步组间两两比较,NS+KD组P25~P31各时间点体重较NS+ND组对应时间点明显降低(P0.05),RS+KD组P23~P31各时间点体重较RS+ND组对应时间点明显降低(P0.05)。2、神经行为学测试:(1)平面翻正实验:与NS+ND组相比,RS+ND组(P27、P31)平面反正时间明显缩短(P0.05);与RS+ND组相比,RS+KD组(P27、P31)平面翻正时间明显延长(P0.05)。(2)悬崖回避实验:与NS+ND组相比,RS+ND组(P27、P31)悬崖回避时间明显延长(P0.05);与RS+ND组相比,RS+KD组(P27、P31)悬崖回避时间明显缩短(P0.05)。(3)前肢悬吊实验:与NS+ND组相比,RS+ND组(P27、P31)前肢悬吊时间明显缩短(P0.05);与RS+ND组相比,RS+KD组(P27、P31)前肢悬吊时间明显延长(P0.05)。(4)负向趋地反应实验:与NS+ND组相比,RS+ND组(P27、P31)负向趋地反应时间明显延长(P0.05);与RS+ND组相比,RS+KD组(P27、P31)负向趋地反应时间明显缩短(P0.05)。3、旷场实验:与NS+ND组相比,RS+ND组延迟时间明显延长(P0.05)、开场得分明显降低(P0.05)、修饰次数明显增多(P0.05);与RS+ND组相比,RS+KD组延迟时间明显缩短(P0.05)、开场得分明显增加(P0.05)、修饰次数明显减少(P0.05)。4、免疫印迹:(1)与NS+ND组相比,RS+ND组海马及大脑皮质Cathepsin E、MT3、CLU的表达明显增高(P0.05);与RS+ND组相比,RS+KD组Cathepsin E、MT3、CLU的表达明显降低(P0.05)。(2)与NS+ND组相比,RS+ND组海马及大脑皮质ZIP7的表达明显降低(P0.05);与RS+ND组相比,RS+KD组ZIP7的表达明显增高(P0.05)。结论1、三氟乙醚诱导新生期大鼠反复长程惊厥可导致生长发育、神经行为及情感认知的损伤,并使海马及大脑皮质远期Cathepsin E、MT3、CLU表达上调,ZIP7表达下调。2、惊厥后给予生酮饮食干预,能改善大鼠的神经行为及认知损伤,并且这种保护作用可能与下调Cathepsin E、MT3、CLU的表达,上调ZIP7的表达有关。3、本研究提示,Cathepsin E、锌离子转运/脂质代谢信号通路可能参与生酮饮食对新生期反复惊厥性脑损伤的保护作用。4、生酮饮食会造成体重降低,但体重下降与神经行为损伤之间没有必然联系。
[Abstract]:Objective To investigate the effect of ketogenic diet on long-term neurobehavioral injury and related molecular expression in neonatal rats with recurrent seizures induced by trifluoroether.Methods Twenty-four Sprague-Dawley (SD) rats aged 8 days were divided into control group (NS group) and convulsion group (RS group) according to the random number table, 12 rats in each group were inhaled. Trifluoroether induced seizures, repeated induction of convulsions for 30 minutes, once a day, the same method for 8 consecutive days; NS group in addition to the non-inhalation of trifluoroether, the rest of the operation is the same. In P21 according to whether to give ketogenic diet (KD) intervention, each group of rats were divided into two groups according to random number table, that is, non-convulsive ordinary diet group (NS + ND group), no convulsion. NS+ND group and RS+ND group were given normal diet, NS+KD group and RS+KD group were given ketogenic diet for 10 days. Blood glucose and serum ketone values were monitored by tail vein of rats in each group on the day of KD implementation, 7 days and 10 days from P9 to the next day. Weight of rats in each group was measured and recorded. Two neurobehavioral tests were performed on P27 and P31, including plane righting, cliff avoidance, forelimb suspension, and negative geotaxis. Results 1. Effect on body weight: Before ketogenic diet intervention, the weight of P17-P20 in RS group was significantly lower than that of NS group at corresponding time points (P Compared with NS+ND group, the body weight of P23-P31 in RS+KD group was significantly decreased (P 0.05). Compared with RS+ND group, the weight of P23-P31 in RS+KD group was significantly decreased (P 0.05). 2. Neurobehavioral test: (1) Plane correction test: Compared with NS+ND group, the planar reverse time of RS+ND group (P 27, P 31) was significantly shortened (P 0.05); Compared with RS+KD group (P 27, P 31), the planar reverse time of RS+ND group (P 27, P 31) was significantly shortened. Positive time was significantly prolonged (P 0.05). (2) Cliff avoidance test: Compared with NS + ND group, RS + ND group (P 27, P 31) cliff avoidance time was significantly prolonged (P 0.05); RS + KD group (P 27, P 31) cliff avoidance time was significantly shortened (P 0.05). (3) Forelimb suspension test: Compared with NS + ND group, RS + ND group (P 27, P 31) forelimb suspension time was significantly shortened (P 0.05); The forelimb suspension time of RS + KD group (P27, P31) was significantly longer than that of NS + ND group (P 0.05). (4) Negative ground reaction test: Compared with NS + ND group, the negative ground reaction time of RS + ND group (P 27, P 31) was significantly longer (P 0.05); Compared with RS + KD group (P 27, P 31), the negative ground reaction time of RS + ND group was significantly shorter (P 0.05). Compared with RS + ND group, RS + KD group had shorter delay time (P 0.05), higher opening score (P 0.05) and fewer modification times (P 0.05). Immunoblotting showed: (1) Compared with NS + ND group, RS + ND group had shorter cathepsin E, MT3 and CLU in hippocampus and cerebral cortex. The expression of ZIP7 in hippocampus and cerebral cortex of RS + ND group was significantly lower than that of RS + ND group (P 0.05). (2) Compared with NS + ND group, the expression of ZIP7 in hippocampus and cerebral cortex of RS + ND group was significantly lower (P 0.05). Compared with RS + ND group, the expression of ZIP7 in RS + KD group was significantly higher (P 0.05). Conclusion 1. Trifluoroether induced repeated long-term shock in neonatal rats. Convulsion can lead to growth and development, neurobehavioral and emotional cognitive impairment, and long-term Cathepsin E, MT3, CLU expression in the hippocampus and cerebral cortex up-regulated, ZIP7 expression down-regulated. 2. Ketogenic diet intervention after convulsion can improve neurobehavioral and cognitive impairment in rats, and this protective effect may be related to down-regulated Cathepsin E, MT3, CLU expression, up-regulated. This study suggests that cathepsin E, zinc ion transport/lipid metabolism signaling pathway may be involved in the protective effect of ketogenic diet on recurrent convulsive brain injury in neonates. 4. Ketogenic diet may cause weight loss, but there is no correlation between weight loss and neurobehavioral injury.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R720.597
本文编号：2240374
[Abstract]:Objective To investigate the effect of ketogenic diet on long-term neurobehavioral injury and related molecular expression in neonatal rats with recurrent seizures induced by trifluoroether.Methods Twenty-four Sprague-Dawley (SD) rats aged 8 days were divided into control group (NS group) and convulsion group (RS group) according to the random number table, 12 rats in each group were inhaled. Trifluoroether induced seizures, repeated induction of convulsions for 30 minutes, once a day, the same method for 8 consecutive days; NS group in addition to the non-inhalation of trifluoroether, the rest of the operation is the same. In P21 according to whether to give ketogenic diet (KD) intervention, each group of rats were divided into two groups according to random number table, that is, non-convulsive ordinary diet group (NS + ND group), no convulsion. NS+ND group and RS+ND group were given normal diet, NS+KD group and RS+KD group were given ketogenic diet for 10 days. Blood glucose and serum ketone values were monitored by tail vein of rats in each group on the day of KD implementation, 7 days and 10 days from P9 to the next day. Weight of rats in each group was measured and recorded. Two neurobehavioral tests were performed on P27 and P31, including plane righting, cliff avoidance, forelimb suspension, and negative geotaxis. Results 1. Effect on body weight: Before ketogenic diet intervention, the weight of P17-P20 in RS group was significantly lower than that of NS group at corresponding time points (P Compared with NS+ND group, the body weight of P23-P31 in RS+KD group was significantly decreased (P 0.05). Compared with RS+ND group, the weight of P23-P31 in RS+KD group was significantly decreased (P 0.05). 2. Neurobehavioral test: (1) Plane correction test: Compared with NS+ND group, the planar reverse time of RS+ND group (P 27, P 31) was significantly shortened (P 0.05); Compared with RS+KD group (P 27, P 31), the planar reverse time of RS+ND group (P 27, P 31) was significantly shortened. Positive time was significantly prolonged (P 0.05). (2) Cliff avoidance test: Compared with NS + ND group, RS + ND group (P 27, P 31) cliff avoidance time was significantly prolonged (P 0.05); RS + KD group (P 27, P 31) cliff avoidance time was significantly shortened (P 0.05). (3) Forelimb suspension test: Compared with NS + ND group, RS + ND group (P 27, P 31) forelimb suspension time was significantly shortened (P 0.05); The forelimb suspension time of RS + KD group (P27, P31) was significantly longer than that of NS + ND group (P 0.05). (4) Negative ground reaction test: Compared with NS + ND group, the negative ground reaction time of RS + ND group (P 27, P 31) was significantly longer (P 0.05); Compared with RS + KD group (P 27, P 31), the negative ground reaction time of RS + ND group was significantly shorter (P 0.05). Compared with RS + ND group, RS + KD group had shorter delay time (P 0.05), higher opening score (P 0.05) and fewer modification times (P 0.05). Immunoblotting showed: (1) Compared with NS + ND group, RS + ND group had shorter cathepsin E, MT3 and CLU in hippocampus and cerebral cortex. The expression of ZIP7 in hippocampus and cerebral cortex of RS + ND group was significantly lower than that of RS + ND group (P 0.05). (2) Compared with NS + ND group, the expression of ZIP7 in hippocampus and cerebral cortex of RS + ND group was significantly lower (P 0.05). Compared with RS + ND group, the expression of ZIP7 in RS + KD group was significantly higher (P 0.05). Conclusion 1. Trifluoroether induced repeated long-term shock in neonatal rats. Convulsion can lead to growth and development, neurobehavioral and emotional cognitive impairment, and long-term Cathepsin E, MT3, CLU expression in the hippocampus and cerebral cortex up-regulated, ZIP7 expression down-regulated. 2. Ketogenic diet intervention after convulsion can improve neurobehavioral and cognitive impairment in rats, and this protective effect may be related to down-regulated Cathepsin E, MT3, CLU expression, up-regulated. This study suggests that cathepsin E, zinc ion transport/lipid metabolism signaling pathway may be involved in the protective effect of ketogenic diet on recurrent convulsive brain injury in neonates. 4. Ketogenic diet may cause weight loss, but there is no correlation between weight loss and neurobehavioral injury.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R720.597
【参考文献】
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1 秦炯;;重视对儿童癫vN共患病的认识与研究[J];中华儿科杂志;2007年08期
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