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目的研究反复惊厥新生大鼠大脑皮质中自噬相关基因微管相关蛋白1轻链3(LC3)的表达及Cathepsin B抑制剂(CBI)对其表达的干预作用。方法 6日龄SD大鼠随机分为3组,每组30只。惊厥组新生大鼠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组惊厥前腹腔注射CBI(总量2μL),采用同样方法吸入三氟乙醚诱导惊厥。3组大鼠于最后一次惊厥后分别按1.5 h、3.0 h、6.0 h、24.0 h和出生35 d(P35)取脑,采用Western blot法分别检测3组大鼠大脑皮质中LC3的表达。结果惊厥组(1.5 h、3.0 h、6.0 h、24.0 h)LC3的表达明显高于对照组同一时间点(Pa<0.05)。CBI组LC3于1.5 h、3.0 h、6.0 h、24.0 h的表达明显低于惊厥组同一时间点(Pa<0.05)。P35时间点3组间LC3水平比较差异无统计学意义。结论 LC3蛋白水平明显上调,表明新生大鼠惊厥后急性期自噬途径被激活,CBI参与了自噬途径的调控。
Objective To investigate the expression of autophagy-related gene 1 light chain 3 (LC3) and the effect of Cathepsin B inhibitor (CBI) on the expression of autophagy in the cerebral cortex of neonatal rats with recurrent seizures. Methods Six-day-old SD rats were randomly divided into three groups of 30 rats. The neonatal rats of convulsion group were infused with trifluoroethyl ether for 5 times to induce seizures every time for 30 min, for 9 consecutive days. In the control group, the same operation was performed but no trifluoroethane was inhaled. The CBI group was injected intraperitoneally with 2 μL ), Using the same method of inhalation of trifluoroethyl ether convulsions. The rats in the three groups were sacrificed at 1.5 h, 3.0 h, 6.0 h, 24.0 h and 35 d after birth (P35) respectively. The expression of LC3 in the cerebral cortex was detected by Western blot. Results The expression of LC3 in the seizure group (1.5 h, 3.0 h, 6.0 h, 24.0 h) was significantly higher than that of the control group at the same time point (Pa <0.05). The expression of LC3 in CBI group at 1.5 h, 3.0 h, 6.0 h and 24.0 h was significantly lower than that at the same time point in the seizure group (P <0.05). There were no significant differences in LC3 levels between the three groups at P35 time point. Conclusion LC3 protein levels were significantly increased, indicating that neonatal rats after acute seizures autophagy pathway is activated, CBI involved in the regulation of autophagy pathway.