| Objectives:Epilepsy is one of the most common neurological diseases that affect nearly1%of the global majority. Moat patients can control the seizures effectively with existing antiepileptic drugs, but there are still20%to40%of people with epilepsy cannot effectively control, called intractable epilepsy. The temporal lobe epilepsy is the most common type of refractory epilepsy,its pathogenesis has always been the hot spot of the epilepsy research.The glutamate acid is the most important excitability amino acid of the central nervous system and its receptor, N-methyl-D-aspartate (NMDA) plays a very important role in the formation and development of epilepsy, especially in temporal lobe epilepsy. The member of the kinesin family17(KIF17) is a kind of kinesin, which was widely expressed in adult vertebrate nervous system.It can transport the NMDA receptor subunit NR2B to the cell membrane along the microtubule,but whether it is involved in fiber budding and synaptic reconstruction of temporal lobe epilepsy remains unknown.To explore its possible role in temporal lobe epilepsy,we investigate the expression of KIF17in the hippocampus and temporal lobe of pilocarpine—induced seizures rats.And we hope we can provides the experimental basis for the clinical treatment of refractory epilepsy.Methods:The animal model was established by lithium-pilocarpine induction in rats. Totally49adult healthy male Wistar rats were randomly divided into control group (n=7) and experimental group (n=42), the experimental group contained six subgroups according to sacrifice time (n=7)(24h,72h,7d,14d,1month, and2months). We examined the expression and localization of KIF17via Western blot and double-label immunofluorescence respectively.Results:1. Behavioral Observation:the epilepsy model of Lithium-Pilocarpine rats develops in three phases:acute phase, silent period and chronic phase. All the rats were similar in behavior after30minutes of pilocarpine-injected, including salivation, mastication,head bobbing,forelimb clonus,posture loss,et al.Of pilocarpine-injected animals,only those that had been observed for continuous convulsion(stage IV or V of Racine) were considered status epilepticus and could be used for further analysis.2. Western blot:In rat hippocampus, optical density ration of KIF17/β-action in the experimental groups were0.5160±0.1961,0.7418±0.3134,0.8876±0.3192,0.7696±0.2713,0.7422±0.2610and0.7144±0.2708,respectively,which were significant differences from control group(0.4948±0.2025,t=7.051,4.974,7.419,8.795,8.264and6.676,all P<0.05).In rat cortex of temporal lobe, optical density ration of KIF17/β-action in the experimental groups were0.4165±0.1599,0.4736±0.1708,0.6365±0.2694,0.6247±0.2221,0.7997±0.3527,0.6523±0.2845,respectively,which were significant differences from control group(0.3447±0.1441, t=5.722,9.061,4.872,7.808,4.791and4.846,all P<0.05).In rat hippocampus KIF17expression increased significantly in the experimental groups especially7d after the onset of seizure using Western bolt. In rat cortex of temporal lobe KIF17expression increased significantly in the experimental groups especially30d after the onset of seizure using Western bolt.3.Double-labeI immunofluorescence:To further asses the cellular localization of KIF17,double-label immunofluorescent stating with microtubule-associate protein2(MAP2),gilia fibrillary acidic protein (GFAP),vesicular glutamate transportersl (BNPI), glutamic acid decarboxylase(GAD67)respectively.We disclosed that KIF17localized in the neurons, Including excitable neurons and inhibitory neurons,but not in astrocytes. Conclusion:l.The expression of KIF17in the brain of pilocarpine-induced seizures in rats,including hippocampus and the cortex of temporal lobe were both higher than the control respectively.2.KIF17localized in the neurons, including excitable neurons and inhibitory neurons,but not in astrocytes.3.KIF17may play a potential role in the pathogenetic mechanisms of the rat lithium-pilocarpine model of epilepsy. |
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