RNA Epigenetic Study Of Anti-epileptic Mechanism Of Valproate Sodium

ObjectiveTo explore the potential mechanism of RNA epigenetic regulation in the treatment of temporal lobe epilepsy with sodium valproate,and provide new clues for the precise treatment of epilepsy.Methods1.A rat model of temporal lobe epilepsy was induced by intraperitoneal injection of lithium chloride-pilocarpine.2.The iTRAQ proteomic analysis of the brain tissue of the epilepsy group and the normal group of rats was performed to find proteins with different expression.3.The spontaneous seizures(SRS)of rats were observed and counted,and the spatial learning and memory ability of rats was detected by water maze.4.Nissl staining and silver staining were used to observe hippocampal neuronal damage and mossy fiber sprouting,respectively.5.Western blot was used to detect the expression levels of methyltransferase,demethylase and differential proteins in brain tissue.6.The m~6A modified expression of differential protein mRNA in brain tissue was detected by MeRIP-qPCR.Results1.The iTRAQ proteomic analysis of brain tissue in the epilepsy group and the normal group was used to find proteins with differential expression.2.VPA can significantly reduce the spontaneous onset of epileptic rats,and can improve the learning and memory ability of epileptic rats.3.Nissl staining can be seen:VPA can significantly improve the cell loss and structural destruction of hippocampal CA1 and CA3 areas in rats with epilepsy.Silver staining found that VPA can cause mossy fiber sprouting in hippocampus of epileptic rats.The increase is significantly inhibited,resulting in a decrease in abnormal synaptic growth.4.In the epileptic group,the Cofilin-1 protein was elevated in the case of normal mRNA expression.VPA treatment can reduce the expression of Cofilin-1 protein.5.There was m~6A hypermethylation modification in the 515 site of Cofilin-1 mRNA in the brain tissue of the epileptic group,but the modification was significantly reduced after VPA treatment.6.There was no significant change in the expression of METTL3 and METTL14 in the methyltransferase complex in each group.The demethylase FTO in the epilepsy group was significantly lower than that in the normal group,and increased after VPA treatment.Conclusions1.Methylation of m~6A in the 515 locus of Cofilin-1 mRNA in the brain tissue of rats with temporal lobe epilepsy can affect the translation process of mRNA and increase the protein expression of Cofilin-1,which is involved in synaptic plasticity and participates in abnormal nerves.The formation of a loop.2.VPA can be involved in the regulation of epigenetic modification of RNA through FTO and exert anti-epileptic effects.