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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 investigate the expression of autophagy marker Beclin-1 in cerebral cortex of rats with recurrent seizure and the effect of lysosome inhibitor E-64d on its expression. Methods Sprague Dawley (SD) rats 21 days old were randomly divided into three groups of 24 rats. Rats in the seizure group were intraperitoneally injected with penicillin (4.8 × 106 U · kg-1 · d-1) every other day for 6 consecutive seizures. The rats in the control group were injected intraperitoneally with the same dose of 9 g · L-1 saline. E-64d Group before each convulsion intraperitoneal injection of 4LE-64d, using the same method of intraperitoneal injection of penicillin convulsion seizures. Three rats were sacrificed at 3h, 12h, 24h after birth and 51d (P51) at the end of the last convulsion. The cerebral cortex was taken and the expression of Beclin-1 in cerebral cortex was detected by Western blot. Application SPSS16.0 software for statistical analysis. Results The expression of Beclin-1 protein at 3h, 12h, 24h in convulsion group was significantly higher than that in the control group at the same time point (Pa <0.05). Beclin-1 protein expression in E-64d group at 3h, 12h, 24h was significantly lower than that in seizure group at the same time point (P <0.05). Beclin-1 protein expression in three groups of P51 was no significant difference (Pa> 0.05). Conclusions The significant increase of Beclin-1 protein in acute phase after convulsions indicates that autophagy pathway is activated and E-64d is involved in the regulation of autophagy pathway.