中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2010年
6期
583-586
,共4页
惊厥%新生期%微管相关蛋白1轻链3%免疫印迹%自噬%3-MA%海马%大鼠
驚厥%新生期%微管相關蛋白1輕鏈3%免疫印跡%自噬%3-MA%海馬%大鼠
량궐%신생기%미관상관단백1경련3%면역인적%자서%3-MA%해마%대서
Seizure%neonatal%LC-3%Western blot%Autophagy%3- MA%Hippocampus%Rat
目的 研究新生期大鼠反复惊厥后海马中自噬标记蛋白微管相关蛋白1轻链3(LC3)的表达及3-MA对表达的干预作用.方法 实验在苏州大学衰老与神经疾病实验室进行.日龄6 d的sprague-Dawley大鼠72只随机(随机数字法)分成三组,每组24只,惊厥组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;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.三组大鼠于最后一次惊厥后分别按1.5 h,3 h,6 h,24 h后取海马组织,采用蛋白质免疫印迹技术(western blot)分别检测三组大鼠大脑海马中LC-3的表达.各组LC3蛋白水平采用方差分析进行比较.结果 对照组和3-MA组同一时间点LC3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LC-3的表达明显高于对照组和3-MA组同一时间点LC3表达(F值分别为4.70,5.28,8.51,5.89,P<0.05).结论 惊厥后自噬途径被激活,3-MA参与了自噬途径的调控.
目的 研究新生期大鼠反複驚厥後海馬中自噬標記蛋白微管相關蛋白1輕鏈3(LC3)的錶達及3-MA對錶達的榦預作用.方法 實驗在囌州大學衰老與神經疾病實驗室進行.日齡6 d的sprague-Dawley大鼠72隻隨機(隨機數字法)分成三組,每組24隻,驚厥組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;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.三組大鼠于最後一次驚厥後分彆按1.5 h,3 h,6 h,24 h後取海馬組織,採用蛋白質免疫印跡技術(western blot)分彆檢測三組大鼠大腦海馬中LC-3的錶達.各組LC3蛋白水平採用方差分析進行比較.結果 對照組和3-MA組同一時間點LC3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LC-3的錶達明顯高于對照組和3-MA組同一時間點LC3錶達(F值分彆為4.70,5.28,8.51,5.89,P<0.05).結論 驚厥後自噬途徑被激活,3-MA參與瞭自噬途徑的調控.
목적 연구신생기대서반복량궐후해마중자서표기단백미관상관단백1경련3(LC3)적표체급3-MA대표체적간예작용.방법 실험재소주대학쇠로여신경질병실험실진행.일령6 d적sprague-Dawley대서72지수궤(수궤수자법)분성삼조,매조24지,량궐조매일흡입삼불을미유도량궐발작5차,매차간격30 min,련속9 d;대조조동양조작단불흡입삼불을미;3-MA조량궐전복강주사3-MA(총량2μL),동양방법흡입삼불을미유도량궐,방법동량궐조.삼조대서우최후일차량궐후분별안1.5 h,3 h,6 h,24 h후취해마조직,채용단백질면역인적기술(western blot)분별검측삼조대서대뇌해마중LC-3적표체.각조LC3단백수평채용방차분석진행비교.결과 대조조화3-MA조동일시간점LC3표체차이무통계학의의,량궐조(1.5 h,3 h,6 h,24 h)해마조직중LC-3적표체명현고우대조조화3-MA조동일시간점LC3표체(F치분별위4.70,5.28,8.51,5.89,P<0.05).결론 량궐후자서도경피격활,3-MA삼여료자서도경적조공.
Objective To explore the repetitive expressions of autophagy marker protein-rnicrotubule-associ-ated protein 1 light chain 3 (LC3) in hippocampus in newborn rats with recurrent seizure and the influence of 3-methyladeine (3-MA) on LC2 expressions. Method Seventy-two 6-day-old SD rats were randomly (random nam-ber) divided into the recurrent neonatal seizure group (RS group, n = 24), the 3-MA-treated seizure group (3-MA group, n = 24) and control group (n = 24). Rats in RS group were subjected to 55 attacks of seizure induced by flurothyl in 9 successive days from the 6th postnatal day (P6). In 3-MA group, 2 μL of 3-MA was injected every day till seizure induced. Western blot analysis was used to determine LC3 protein level in hippocampus at different intervals of 1.5 h,3 h,6 h and 24 h after the last convulsion. The LC3 protein level was analyzed with Dunnett test after ANOVA. Results LC3 protein levels in RS group at the different intervals were significantly higher than those in the control group and in 3-MA group (F =4.70,5.28,8.51 and 5.89, respectively, P <0.05), and there were no significant differences in LC3 protein level between 3-MA group and control group at those intervals (P > 0.05). Conclusions The autophagy/lysosomal pathway is immediately activated after recurrent seizure evidenced by the elevated expressions of LC3 in hippocampus. The 3-MA is involved in the regulation of autophagy/ lysosomal pathway by down-regulating the expressions of LC3.
