中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2011年
3期
255-258
,共4页
惊厥%Beclin-1%Cathepsin B%Bcl-2%自噬%CBI
驚厥%Beclin-1%Cathepsin B%Bcl-2%自噬%CBI
량궐%Beclin-1%Cathepsin B%Bcl-2%자서%CBI
Seizure%Beclin-1%Cathepsin B%Bcl-2%autophagy%CBI
目的 探讨新生期大鼠反复惊厥后海马自噬、凋亡相关蛋白Beclin-1,Cathepsin B和Bcl-2的表达及溶酶体蛋白酶抑制剂CBI对表达的干预作用.方法 实验在苏州大学神经科学研究所进行.日龄6 d的Sprague-Dawley大鼠90只随机(随机数字法)分为3组,每组30只:对照组、反复惊厥组及CBI组.反复惊厥组(RS)每日吸入三氟乙醚诱导惊厥发作5次,每次间隔30 min,连续9 d;对照组同样操作但不吸入三氟乙醚;CBI组于惊厥前腹腔注射cathepsin B(2 μL,0.5μg/μL),同样方法吸入三氟乙醚诱导惊厥发作.各组分别于末次惊厥后1.5 h,3 h,6 h,24 h和生后35 d 5个时间点[(n=6/(每点,每组)]取海马组织,采用免疫印迹技术检测自噬相关蛋白Beclin-1,Cathepsin B和Bcl-2的表达,采用方差分析进行统计.结果 对照组和CBI组同一时间点Beclin-1,Cathepsin B和Bcl-2的表达差异无统计学意义,惊厥组(1.5 h,3 h,6 h,24 h)海马组织中Beclin-1和Cathepsin B的表达明显高于对照组和CBI组同一时间点的Beclin-1表达(F值分别为Beclin-1:7.44,6.58,5.43,9.69,P＜0.05;Cathepsin B:4.62,2.02,3.49,4.86);而惊厥组(1.5 h,3 h,6 h,24 h)Bcl-2的表达则明显低于对照组和CBI组同一时间点Bcl-2的表达(F值分别为4.34,3.79,4.88,4.42,P＜0.05).结论 发育期大鼠反复惊厥后海马神经元自噬信号通路激活,同时抗凋亡基因Bcl-2蛋白水平显著下调,表明自噬信号途径与凋亡途径共同激活,协调参与发育期大鼠惊厥急性期兴奋毒性脑损伤的病理生理机制.
目的 探討新生期大鼠反複驚厥後海馬自噬、凋亡相關蛋白Beclin-1,Cathepsin B和Bcl-2的錶達及溶酶體蛋白酶抑製劑CBI對錶達的榦預作用.方法 實驗在囌州大學神經科學研究所進行.日齡6 d的Sprague-Dawley大鼠90隻隨機(隨機數字法)分為3組,每組30隻:對照組、反複驚厥組及CBI組.反複驚厥組(RS)每日吸入三氟乙醚誘導驚厥髮作5次,每次間隔30 min,連續9 d;對照組同樣操作但不吸入三氟乙醚;CBI組于驚厥前腹腔註射cathepsin B(2 μL,0.5μg/μL),同樣方法吸入三氟乙醚誘導驚厥髮作.各組分彆于末次驚厥後1.5 h,3 h,6 h,24 h和生後35 d 5箇時間點[(n=6/(每點,每組)]取海馬組織,採用免疫印跡技術檢測自噬相關蛋白Beclin-1,Cathepsin B和Bcl-2的錶達,採用方差分析進行統計.結果 對照組和CBI組同一時間點Beclin-1,Cathepsin B和Bcl-2的錶達差異無統計學意義,驚厥組(1.5 h,3 h,6 h,24 h)海馬組織中Beclin-1和Cathepsin B的錶達明顯高于對照組和CBI組同一時間點的Beclin-1錶達(F值分彆為Beclin-1:7.44,6.58,5.43,9.69,P＜0.05;Cathepsin B:4.62,2.02,3.49,4.86);而驚厥組(1.5 h,3 h,6 h,24 h)Bcl-2的錶達則明顯低于對照組和CBI組同一時間點Bcl-2的錶達(F值分彆為4.34,3.79,4.88,4.42,P＜0.05).結論 髮育期大鼠反複驚厥後海馬神經元自噬信號通路激活,同時抗凋亡基因Bcl-2蛋白水平顯著下調,錶明自噬信號途徑與凋亡途徑共同激活,協調參與髮育期大鼠驚厥急性期興奮毒性腦損傷的病理生理機製.
목적 탐토신생기대서반복량궐후해마자서、조망상관단백Beclin-1,Cathepsin B화Bcl-2적표체급용매체단백매억제제CBI대표체적간예작용.방법 실험재소주대학신경과학연구소진행.일령6 d적Sprague-Dawley대서90지수궤(수궤수자법)분위3조,매조30지:대조조、반복량궐조급CBI조.반복량궐조(RS)매일흡입삼불을미유도량궐발작5차,매차간격30 min,련속9 d;대조조동양조작단불흡입삼불을미;CBI조우량궐전복강주사cathepsin B(2 μL,0.5μg/μL),동양방법흡입삼불을미유도량궐발작.각조분별우말차량궐후1.5 h,3 h,6 h,24 h화생후35 d 5개시간점[(n=6/(매점,매조)]취해마조직,채용면역인적기술검측자서상관단백Beclin-1,Cathepsin B화Bcl-2적표체,채용방차분석진행통계.결과 대조조화CBI조동일시간점Beclin-1,Cathepsin B화Bcl-2적표체차이무통계학의의,량궐조(1.5 h,3 h,6 h,24 h)해마조직중Beclin-1화Cathepsin B적표체명현고우대조조화CBI조동일시간점적Beclin-1표체(F치분별위Beclin-1:7.44,6.58,5.43,9.69,P＜0.05;Cathepsin B:4.62,2.02,3.49,4.86);이량궐조(1.5 h,3 h,6 h,24 h)Bcl-2적표체칙명현저우대조조화CBI조동일시간점Bcl-2적표체(F치분별위4.34,3.79,4.88,4.42,P＜0.05).결론 발육기대서반복량궐후해마신경원자서신호통로격활,동시항조망기인Bcl-2단백수평현저하조,표명자서신호도경여조망도경공동격활,협조삼여발육기대서량궐급성기흥강독성뇌손상적병리생리궤제.
Objective To explore the dynamic expressions of autophagia and apoptosis associated protein Beclin-1, Cathepsin B and Bcl-2 in hippocampus and the intervention efficacy of cathepsin-B inhibitor (CBI) after recurrent neonatal seizure. Method Ninty 6-day-old SD rats were randomly (random number) divided into the recurrent neonatal seizure group (RS group, n = 30), CBI-treated seizure group (CBI group, n = 30) and the control group (n = 30). Rats in RS group were subjected to 55 attacks of seizure induced by using flurothyl during the consecutive 9 days beginning on the 6 th postnatal day (P6). In CBI group, CBI (2 μL, 0.5 μg/μL) was administered every day before seizures induced. Western blot was employed to determine the protein level at different intervals (1.5 h,3 h,6 h,24 h) after the last convulsion.Results There were higher expressions of Beclin-1 and Cathepsin B, and lower expressions of Bcl-2 expression in RS group(1. 5 h,3 h,6 h and 24 h) than those at the same time in control group (P ＜ 0.05). Beclin1 and Cathepsin B expressions were lower while Bcl-2 expressions were higher in CBI group at the intervals of 1.5 h,3 h,6 h and 24 h compared with those in RS group (P ＜ 0. 05). Conciusions Autophagic and apoptotic pathways were activated immediately after recurrent neonatal seizures as indicated by expression changes of Beclin-1, Cathepsin B and Bcl-2 in hippocampus, which suggests a synergistic effect of the two pathways in the pathophysiology of the long-term brain damage of neonateε resulted from the adverse effects of recurrent neonatal seizures.
