The Study On Anticonvulsant Effects Of Gentiopicroside And Its Mechanism Involved In NR2B/CaMKⅡ/CREB On Pilocarpine Induced Epileptic Seizures In Mice

OBJECTIVE: Epilepsy is defined as a common and major brain disorder,and one-third of individuals still experience seizures that do not respond to medication.Gentiopicroside(Gent)was found to possess N-methyl-d-aspartate(NMDA)receptor modulatory effect.In this study,we investigate the anticonvulsant and neuroprotective effects of Gent against Pilocarpine(Pilo)Induced epileptic seizures.MATERIALS AND METHODS: Thirty minutes prior to the Pilocarpine(Pilo)injection,the mice were administrated with Gent once.We observed seizure activity and electroencephalography(EEG)after the Pilo injection,and mice were killed for Nissl and Fluoro-jade B(FJB)staining at 72 h after convulsion or SE onset.The expression of the Bax,Bcl-2,caspase-3,NR2 B,calcium/calmodulin-dependent protein kinase Ⅱ(CaMKⅡ)and c AMP response element-binding protein(CREB)was also evaluated in the hippocampus region of the brain by using western blot and q-PCR at 72 h after convulsion or SE onset.RESULTS: In this study,we found that pretreatment with Gent attenuated Pilo-induced convuulsions and mortality in mice as well neurodegeneration in the cells.Analysis of EEG recordings revealed that Gent significantly reduced polyspike and epileptiform discharges.Besides,western blot and q-PCR analysis showed that Gent significantly reversed NR2 B over-expression,decreased CaMKⅡ and CREB respectively.Moreover,Pilo decreased Bcl-2,increased Bax expression and finally the activation of caspase-3.Gent pretreatment reversed all these deleterious effects induced by Pilo.CONCLUSION: Collectively,the findings of this study indicated that Gent exerts anticonvulsant and neuroprotective effects via activation of NR2B/CaMKⅡ/CREB pathway and suggest a potential therapeutic role in epilepsy.