摘要
目的探讨反复惊厥发育期大鼠大脑皮质中自噬标记蛋白Beclin-1的表达及溶酶体酶抑制剂E-64d对其表达的干预作用。方法将出生21d的SD大鼠随机分成3组,每组24只。惊厥组大鼠隔日腹腔注射青霉素(4.8×106U·kg-1·d-1),连续6次,诱导惊厥发作;对照组大鼠予同等剂量的9g·L-1盐水腹腔注射;E-64d组于每次惊厥前腹腔注射4μLE-64d,采用同样方法腹腔注射青霉素诱导惊厥发作。于末次惊厥后3h、12h、24h和出生51d(P51)处死3组大鼠,并取其大脑皮质,采用免疫印迹技术分别检测3组大鼠大脑皮质中Beclin-1蛋白的表达。应用SPSS16.0软件进行统计学分析。结果惊厥组大鼠3h、12h、24h Beclin-1蛋白的表达均明显高于对照组同一时间点Beclin-1蛋白的表达(Pa<0.05)。E-64d组3h、12h、24h Beclin-1蛋白的表达明显低于惊厥组同一时间点Beclin-1蛋白的表达(Pa<0.05)。3组P51时Beclin-1蛋白表达比较差异均无统计学意义(Pa>0.05)。结论惊厥后急性期Beclin-1蛋白水平明显上调表明自噬途径被激活,且E-64d参与自噬途径的调控。
Objective To explore the dynamic expression 6f autophagy - associated protein Beclin - 1 in cerebral cortex in developmental period rats with recurrent seizure（RS） and the intervention effect of E - 64d on Beclin - 1 expression. Methods Twenty - one - day - old （ P21 ） SD rats were randomly divided into 3 groups （24 cases per group） . Rats in RS group were injected i. p. with penicillin（4.8×106 U·kg-1·d-1 ）every other day in eleven consecutive days starting from postnatal day 21st to induce seizure;Rats in the control group were injected with equal amount of normal sodium at the same time;In E -64d group,4 μL E -64d was injected each day before seizure was induced. Western blot analysis was used to determine Beclin - 1 levels in cerebral cortex at different time points （ 3 h, 12 h ,24 h and P51 ） after the last convulsion. SPSS 16.0 software was used to analyze the data. Restflts Beclin - 1 levels in RS group at 3 h, 12 h ,24 h were significantly higher than those at the corresponding time points in the control group （P, 〈 0. 05 ）. The protein levels in E -64d group at the time points of 3 h, 12 h,24 h were decreased significantly compared with those in RS group （P〈 0.05 ）. There were no significant differences among 3 groups at P51 （P 〉 0.05）. Conclusions Autophagy/lysosomal pathway are activated immediately after RS as indicated by the elevated expression of Beclin - 1 in cerebral cortex. E - 64d is involved in the regulation of autophagy/lysosomal pathway.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2010年第23期1828-1830,共3页
Journal of Applied Clinical Pediatrics
基金
国家自然科学基金(30870808)
江苏省自然科学基金青年科技创新人才项目(BK2007509)
