Analysis on the effects of Tianma Gouteng Decoction on miRNA-9 expression in hippocampus of epileptic mice and its protective mechanism against nerve injury
Objective:To investigate the changes of electroencephalogram(EEG),expression level of miRNA-9 in hippocampus of mice and protective mechanism against nerve injury after Tianma Gouteng Decoction intervention in epileptic mice.Methods:Fifty male Kunming mice were randomly divided into control group,model group,Carbamazepine group,TCM high-dose group and TCM low-dose group,with 10 mice in each group.One week before modeling,both control group and model group were given 20 mL·kg1·d-1 normal saline for intragastric administration;Carbamazepine was given 30 mg·kg-1·d-1 Carbamazepine suspension by intragastric administration.Tianma Gouteng Decoction was used to administer 30 mL·kg 1·d-1 and 10 mL·kg-1·d-1 to high dose group and low dose group,respectively.The acute epilepsy model mice was established by pilocarpine hydrochloride.Behavior and EEG were observed in each group.The expression of miRNA-9 in the hippocampus of mice was detected by real-time fluorescence quantitative PCR.Immunohistochemistry was used to detect neuron specific nuclear protein(NeuN)-labeled neuron nuclear antigen,FJB marks damaged neurons and platelet endothelial cell adhesion molecule-1(PECAM-1)-labeled platelet endothelial cell adhesion molecule-1,tight connexin-1(ZO-1)-labeled cytoplasmic tight adhesive protein 1.The level of ZO-1/ZONAB was determined by Western Blot.Results:Compared with the control group,model group mice seizure frequency,duration and seizure level were high(P＜0.01),electroencephalogram in mice sharp wave,spike wave,spine slow wave obviously increased,the expression of the hippocampus of miRNA-9 significantly lowered(P＜0.01);Compared with model group,high dose group and low dose group mice seizure frequency,duration and seizure level were reduced(P＜0.05),mice EEG epileptiform wave decreases,the expression of miRNA-9 was increased in the hippocampus of mice(P＜0.01).Compared with model group,NeuN cells in Carbamazepine group,high-dose group and low-dose group were significantly increased(P＜0.05),and damaged neurons were significantly decreased(P＜0.05).Pecam-1 immune response in hippocampal CA3 region was significantly decreased in high-dose group and low-dose group(P＜0.01).ZO-1 protein level was significantly decreased(P＜0.05),while ZONAB protein level was significantly increased(P＜0.05).Conclusion:Tianma Gouteng Decoction can inhibit neuronal damage induced by epilepsy and has a protective effect on the stability of blood-brain barrier,which may be mediated by ZO-1/ZONAB signaling pathway,so as to play an antiepileptic role.
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