中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2011年
9期
935-939
,共5页
惊厥%新生期%Bcl-2%3-MA%海马
驚厥%新生期%Bcl-2%3-MA%海馬
량궐%신생기%Bcl-2%3-MA%해마
Seizure%Neonatal%Bcl-2%3-MA%Hippecampus
目的 探讨大鼠新生期反复惊厥急性期应用自噬抑制剂3-甲基腺嘌呤(3-MA)对惊厥所致神经行为损伤的干预作用及机制。方法 实验在苏州大学衰老与神经疾病实验室进行。日龄6d(P6,下同)的Sprague-Dawley大鼠45只随机(随机数字法)分成三组,每组15只,惊厥组在P6吸入三氟乙醚诱导惊厥发作,持续30 min,连续6d;对照组同样操作但不吸入三氟乙醚;3-MA组惊厥前腹腔注射3 -MA(总量2μL),同样方法吸入三氟乙醚诱导惊厥,方法同惊厥组。三组大鼠于P12进行游泳行为评分测试大鼠神经运动发育,P17进行旷场实验,P43～P49行Morris水迷宫测试大鼠学习记忆功能,于PS0取海马组织,采用免疫印迹技术(Western blot)检测抗凋亡基因Bcl-2及自噬标记蛋白Beclin1的表达。各组行为学指标和蛋白水平采用方差分析进行统计。结果 惊厥后各组大鼠游泳行为测评,各项评分惊厥组明显低于对照组及3-MA组(P＜0.01),旷场实验显示惊厥组延迟时间[(13.33±6.69)8]较对照组[(7.11±2.37)8]和3-MA组[(9.91±4.23)s]显著延长,差异具有统计学意义(F=4.39,P＜0.05);Morris水迷宫测试RS组第4、5天逃避潜伏期较对照组和3-MA组明显延长(P＜0.05)。惊厥组海马Bcl-2表达水平(0.587±0.139)较对照组(0.782±0.083)及3-MA组(0.799±0.163)显著降低,差异具有统计学意义(F=4.71,P＜0.05)。各组Beclin1的表达差异无统计学意义(F =0.27,P＞0.05)。结论 3-MA能显著改善大鼠惊厥后神经行为损伤,并可能与上调海马抗凋亡基因Bcl-2表达有关。
目的 探討大鼠新生期反複驚厥急性期應用自噬抑製劑3-甲基腺嘌呤(3-MA)對驚厥所緻神經行為損傷的榦預作用及機製。方法 實驗在囌州大學衰老與神經疾病實驗室進行。日齡6d(P6,下同)的Sprague-Dawley大鼠45隻隨機(隨機數字法)分成三組,每組15隻,驚厥組在P6吸入三氟乙醚誘導驚厥髮作,持續30 min,連續6d;對照組同樣操作但不吸入三氟乙醚;3-MA組驚厥前腹腔註射3 -MA(總量2μL),同樣方法吸入三氟乙醚誘導驚厥,方法同驚厥組。三組大鼠于P12進行遊泳行為評分測試大鼠神經運動髮育,P17進行曠場實驗,P43～P49行Morris水迷宮測試大鼠學習記憶功能,于PS0取海馬組織,採用免疫印跡技術(Western blot)檢測抗凋亡基因Bcl-2及自噬標記蛋白Beclin1的錶達。各組行為學指標和蛋白水平採用方差分析進行統計。結果 驚厥後各組大鼠遊泳行為測評,各項評分驚厥組明顯低于對照組及3-MA組(P＜0.01),曠場實驗顯示驚厥組延遲時間[(13.33±6.69)8]較對照組[(7.11±2.37)8]和3-MA組[(9.91±4.23)s]顯著延長,差異具有統計學意義(F=4.39,P＜0.05);Morris水迷宮測試RS組第4、5天逃避潛伏期較對照組和3-MA組明顯延長(P＜0.05)。驚厥組海馬Bcl-2錶達水平(0.587±0.139)較對照組(0.782±0.083)及3-MA組(0.799±0.163)顯著降低,差異具有統計學意義(F=4.71,P＜0.05)。各組Beclin1的錶達差異無統計學意義(F =0.27,P＞0.05)。結論 3-MA能顯著改善大鼠驚厥後神經行為損傷,併可能與上調海馬抗凋亡基因Bcl-2錶達有關。
목적 탐토대서신생기반복량궐급성기응용자서억제제3-갑기선표령(3-MA)대량궐소치신경행위손상적간예작용급궤제。방법 실험재소주대학쇠로여신경질병실험실진행。일령6d(P6,하동)적Sprague-Dawley대서45지수궤(수궤수자법)분성삼조,매조15지,량궐조재P6흡입삼불을미유도량궐발작,지속30 min,련속6d;대조조동양조작단불흡입삼불을미;3-MA조량궐전복강주사3 -MA(총량2μL),동양방법흡입삼불을미유도량궐,방법동량궐조。삼조대서우P12진행유영행위평분측시대서신경운동발육,P17진행광장실험,P43～P49행Morris수미궁측시대서학습기억공능,우PS0취해마조직,채용면역인적기술(Western blot)검측항조망기인Bcl-2급자서표기단백Beclin1적표체。각조행위학지표화단백수평채용방차분석진행통계。결과 량궐후각조대서유영행위측평,각항평분량궐조명현저우대조조급3-MA조(P＜0.01),광장실험현시량궐조연지시간[(13.33±6.69)8]교대조조[(7.11±2.37)8]화3-MA조[(9.91±4.23)s]현저연장,차이구유통계학의의(F=4.39,P＜0.05);Morris수미궁측시RS조제4、5천도피잠복기교대조조화3-MA조명현연장(P＜0.05)。량궐조해마Bcl-2표체수평(0.587±0.139)교대조조(0.782±0.083)급3-MA조(0.799±0.163)현저강저,차이구유통계학의의(F=4.71,P＜0.05)。각조Beclin1적표체차이무통계학의의(F =0.27,P＞0.05)。결론 3-MA능현저개선대서량궐후신경행위손상,병가능여상조해마항조망기인Bcl-2표체유관。
Objective To explore the intervention effect and the underlying molecular mechanism of 3-Methyladenine on behavioral damage of neonatal rat with prolonged seizures. Methods Forty-five 6-dayold SD rats were randomly ( random number) divided into the recurrent prolonged neonatal seizure group ( RS group), the 3-MA-treated seizure group and control group. The volatile agent flurothyl was used to induce 30 min seizure attack. At postnatal day 6 (P6), recurrent seizures were induced once daily for successive 6 days in both RS group and 3-MA group. In 3-MA group, 3-MA (2 μL) was injected daily before seizures induced. Neural-behavior changes were observed with double-blind method including swimming development, open field test and Morris water maze analysis. Bcl-2 and Beclinl protein levels in hippocampus were detected by western blot method at P50. Results The total scores of swimming behavior in RS rats were decreased significantly compared with those of control and 3-MA rats ( control: 7. 44 ±1. 13, RS: 5.06±1.63, 3-MA: 7.33 ±1.08, F=16.19, P＜0. 01) . The start-latency time of open filed behavior in RS rats ( 13. 33 ±6. 69) were increased significantly compared with that of control (7. 11 ±2. 37) and 3-MA rats (9. 91 ±4. 23) (F=4. 39, P＜0. 05). The escape latency was significantly longer in rats of RS group than that of control and 3-MA rats on the 4th and 5th days (P ＜ 0.05). The level of Bcl-2 in hippocampus of RS group (0. 587 +0. 139) were significantly lower than that of control (0. 782 +0. 083) and 3-MA groups (0. 799 + 0. 163) (F =4. 7 1, P ＜ 0. 05 ). There were no significant differences in the level of Beclin1 protein in hippocampus among the three groups ( F =0. 27, P ＞ 0. 05 ). Conclusions Pretreatment with autophagy inhibitor 3-MA in acute phase of neonatal seizures could significantly improve neurobehavioral capacity, which might be associated with the increased in the level of Bcl-2 protein in hippocampus.
