PTZ诱导的癫痫急性发作大鼠大脑神经元中F-actin、Calponin3和ROCK2的表达及其意义

doi:10.13481/j.1671-587x.20150218

Abstract

Abstract:

Objective To disscuss the effect of the acute epileptic seizure induced by pentetrazol (PTZ) on the expressions of F-actin,Calponin3,and ROCK2 in the cerebral neurons of the rats,and to illuminate the possible mechanism of preventing the F-actin from abnormal depolymerization and triggering the rearrangement of F-actin.Methods 56 immature rats aged 3 weeks were divided into control (n=6) and epilepsy (n=50) groups. The rats in control group were injected with physiological saline introperitoneally.The rats in epilepsy group were introperitoneally injected with 60 mg·kg^-1 PTZ to establish the acute epileptic seizure models.And 6 rats from successful acute epileptic seizure models were sacrificed at different time points (1,2,3,and 7 d) after modeling.The fluorescence intensity of F-actin in the internal molecular layer of hippocampus of the rats was observed by phalloidine staining labeled by Alex-488; the distributions of Calponin3 and ROCK2 in the cerebral neurons in the pallium and hippocampus of the rats were detected by immunofluorescence method; the relative expression levels of Calponin3,ROCK2,and phosphorylated ROCK2 were analyzed by Western blotting method.Results Compared with control group,the fluorescence intensity of F-actin in the hippocampal internal molecular layer of the rats in epilepsy group was decreased (P<0.05),and the dot-shaped aggregation of F-actin was disappeared 1 d after modeling.The immunofluorescence results showed that Calponin3 dispersed in the cytoplasm of the neurons of the rats in control group;however,it aggregated in the cell cortex of the neurons 7 d after modeling in epilepsy group.ROCK2 was located in a small amount of neuritis of the rats in control group,whereas a great quantity of ROCK2 was found in both of the cell body and neuritis in epilepsy group 7 d after modeling.The Western blotting results showed that the relative expression levels of Calponin3 protein in the epilepsy group were markedly decreased at different time points after modeling compared with control group (P<0.05);however,it was increased gradually with the prolongation of time and closed to the level in control group within 1 week.The relative expression levels of ROCK2 protein in the hippocampus of the rats in epilepsy group began to increase steadily from 3 to 7 d after modeling compared with control group (P<0.05);however,the relative expression levels of phosphorylated ROCK2 protein in epilepsy group were increased sustainably from 1 to 7 d after modeling compared with control group (P<0.05),and were decreased gradually in a time-dependent manner.Conclusion The acute epileptic seizure of the immature rats induced by PTZ not only leads to the abnormal depolymerization of F-actin,but also actives the RhoA/ROCK2 signal pathway simultaneously and up-regulates the expression levels of ROCK2 and Calponin3 proteins.

Key words: epilepsy, pentetrazol, filiment-actin, Calponin3, ROCK2

CLC Number:

R742.1

CHENG Chunxu, LI Shulei, GU Xiaoyun, HUANG Kexin, LI Yanchao, ZHANG Shuyan, YANG Libin. Expressions of F-actin,Calponin3,and ROCK2 in cerebral neurons of rats with acute epileptic seizure induced by PTZ[J].Journal of Jilin University Medicine Edition, 2015, 41(02): 299-303.

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Expressions of F-actin,Calponin3,and ROCK2 in cerebral neurons of rats with acute epileptic seizure induced by PTZ

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