目的研究新生期大鼠反复惊厥后海马中Beclin-1急性期的表达及3-甲基腺嘌呤f3-MA)的干预作用。方法生后6d的SD大鼠108只按照随机数字表法分成3组,每组36只,惊厥组每日吸入三氟乙醚诱导惊厥发作5次,每次间隔30min,连续9d;对照组同样操作但不吸人三氟乙醚;3-MA组惊厥前腹腔注射3-MA(总量2μL),然后以同样方法吸入三氟乙醚诱导惊厥,方法同惊厥组。3组大鼠于最后一次惊厥后1.5h、3h、6h、12h、24h、48h断头取脑,每组每个时间点分别为6只大鼠,采用Western blot分别检测3组大鼠大脑海马中Beclin-1的表达。结果与对照组同一时间点相比,惊厥组海马组织中Beclin-1的表达明显增多,差异均有统计学意义(P〈0.05)。与惊厥组同一时间点相比,3-MA组Beclin-1的表达明显减少,差异均有统计学意义(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。
Objective To study the role of autophagy/lysosomal pathway in developmental seizure-induced apoptosis of the hippocampal cells, we investigated the dynamic expression of Beclin-1 in the hippocampus and the intervention effect of autophagy/lysosome inhibitor 3-methyladenine (3-MA) on Beclin-1 expression following recurrent seizures in neonatal rats. Methods One hundred and eight SD rats aged 6 d were randomly divided into 3 groups: repeated neonatal seizures (RS group), 3-MA-treated seizure (3-MA group) and the control grous(n=36). Rats in the RS group were performed the indraft of flurothyl 5 times daily for a consecutive 9 d. In the 3-MA group, 3-MA was injected every day with a total dosage of 2 μL before seizures were induced. The rats in the 3 groups were sacrificed at 1.5, 3, 6, 12, 24 and 48 h after the last convulsion with 6 rats at each time points; Western blotting was employed to determine the expression of Beclin-1 in the hippocampus at these different time points. Results The expression of Beclin-1 in the RS group was significantly higher than that in the control group at the same time points (P〈0.05). The expression of Beclin-1 at each time point (1.5, 3, 6, 12, 24 and 48 h) in the 3-MA group was significantly decreased compared with that in the RS group (P〈0.05). Conclusion Autophagy/lysosomal pathway are activated immediately after the neonatal seizures and 3-MA involves in the regulation of autophagy/lysosomal pathway by down-regulating the expression of Beclin- 1.