生酮飲食對新生期反復(fù)驚厥大鼠遠(yuǎn)期神經(jīng)行為損傷及相關(guān)分子表達(dá)的影響
發(fā)布時間：2018-09-13 06:38
【摘要】：目的探討生酮飲食對三氟乙醚誘導(dǎo)的新生期反復(fù)驚厥大鼠遠(yuǎn)期神經(jīng)行為損傷及相關(guān)分子表達(dá)的影響。方法日齡8天(用P8表示,下同)的Sprague-Dawley(SD)大鼠24只,按照隨機(jī)數(shù)字表法分為對照組(NS組)和驚厥組(RS組),每組各12只。RS組大鼠吸入三氟乙醚誘導(dǎo)驚厥發(fā)作,反復(fù)誘導(dǎo)驚厥持續(xù)30分鐘,每日一次,相同方法連續(xù)誘導(dǎo)8天;NS組除不吸入三氟乙醚其余操作相同。于P21依據(jù)是否給予生酮飲食(KD)干預(yù),每組大鼠按照隨機(jī)數(shù)字表法再分為兩組,即未驚厥普通飲食組(NS+ND組)、未驚厥生酮飲食組(NS+KD組)、驚厥普通飲食組(RS+ND組)、驚厥生酮飲食組(RS+KD組),每組各6只。NS+ND組和RS+ND組給予普通飲食,NS+KD組和RS+KD組給予生酮飲食,飲食干預(yù)10天。于KD實施當(dāng)天、7天、10天每組大鼠尾靜脈采血監(jiān)測血糖及血酮值。自P9起隔天測量并記錄每組大鼠的體重。于P27、P31分別進(jìn)行兩次神經(jīng)行為學(xué)測試,包括平面翻正、懸崖回避、前肢懸吊、負(fù)向趨地反應(yīng)實驗。P31行曠場實驗。于P32大鼠斷頭取海馬及大腦皮質(zhì),采用免疫印跡法(western blot)檢測各組大鼠海馬及大腦皮質(zhì)中Cathepsin E、ZIP7、MT3、CLU蛋白的表達(dá)。結(jié)果1、對體重的影響:生酮飲食干預(yù)前,RS組P17~P20各時間點體重較NS組對應(yīng)時間點明顯降低(P0.05)。生酮飲食干預(yù)期間(P21~P31),四組大鼠的體重變化存在總體性差異(P0.05),進(jìn)一步組間兩兩比較,NS+KD組P25~P31各時間點體重較NS+ND組對應(yīng)時間點明顯降低(P0.05),RS+KD組P23~P31各時間點體重較RS+ND組對應(yīng)時間點明顯降低(P0.05)。2、神經(jīng)行為學(xué)測試:(1)平面翻正實驗:與NS+ND組相比,RS+ND組(P27、P31)平面反正時間明顯縮短(P0.05);與RS+ND組相比,RS+KD組(P27、P31)平面翻正時間明顯延長(P0.05)。(2)懸崖回避實驗:與NS+ND組相比,RS+ND組(P27、P31)懸崖回避時間明顯延長(P0.05);與RS+ND組相比,RS+KD組(P27、P31)懸崖回避時間明顯縮短(P0.05)。(3)前肢懸吊實驗:與NS+ND組相比,RS+ND組(P27、P31)前肢懸吊時間明顯縮短(P0.05);與RS+ND組相比,RS+KD組(P27、P31)前肢懸吊時間明顯延長(P0.05)。(4)負(fù)向趨地反應(yīng)實驗:與NS+ND組相比,RS+ND組(P27、P31)負(fù)向趨地反應(yīng)時間明顯延長(P0.05);與RS+ND組相比,RS+KD組(P27、P31)負(fù)向趨地反應(yīng)時間明顯縮短(P0.05)。3、曠場實驗:與NS+ND組相比,RS+ND組延遲時間明顯延長(P0.05)、開場得分明顯降低(P0.05)、修飾次數(shù)明顯增多(P0.05);與RS+ND組相比,RS+KD組延遲時間明顯縮短(P0.05)、開場得分明顯增加(P0.05)、修飾次數(shù)明顯減少(P0.05)。4、免疫印跡:(1)與NS+ND組相比,RS+ND組海馬及大腦皮質(zhì)Cathepsin E、MT3、CLU的表達(dá)明顯增高(P0.05);與RS+ND組相比,RS+KD組Cathepsin E、MT3、CLU的表達(dá)明顯降低(P0.05)。(2)與NS+ND組相比,RS+ND組海馬及大腦皮質(zhì)ZIP7的表達(dá)明顯降低(P0.05);與RS+ND組相比,RS+KD組ZIP7的表達(dá)明顯增高(P0.05)。結(jié)論1、三氟乙醚誘導(dǎo)新生期大鼠反復(fù)長程驚厥可導(dǎo)致生長發(fā)育、神經(jīng)行為及情感認(rèn)知的損傷,并使海馬及大腦皮質(zhì)遠(yuǎn)期Cathepsin E、MT3、CLU表達(dá)上調(diào),ZIP7表達(dá)下調(diào)。2、驚厥后給予生酮飲食干預(yù),能改善大鼠的神經(jīng)行為及認(rèn)知損傷,并且這種保護(hù)作用可能與下調(diào)Cathepsin E、MT3、CLU的表達(dá),上調(diào)ZIP7的表達(dá)有關(guān)。3、本研究提示,Cathepsin E、鋅離子轉(zhuǎn)運(yùn)/脂質(zhì)代謝信號通路可能參與生酮飲食對新生期反復(fù)驚厥性腦損傷的保護(hù)作用。4、生酮飲食會造成體重降低,但體重下降與神經(jīng)行為損傷之間沒有必然聯(lián)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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R720.597
本文編號：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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R720.597
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相關(guān)期刊論文 前1條
1 秦炯;;重視對兒童癲vN共患病的認(rèn)識與研究[J];中華兒科雜志;2007年08期
，本文編號：2240374
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