The Protective Effect Of Fasudil On The Irreversible Injury Of Dendritic Spine Caused By Epilepsy

Previous studies had found that patients with epilepsy can cause cognitive dysfunction.According to current treatment methods for epilepsy,there is no well treatment for the impairment of learning and memory function caused by epilepsy.Current research shows that the change in the plasticity of dendritic spines is the basis of learning and memory.Our lab also found that mouse dendritic edema and irreversible damage to dendritic spines would occur after acute seizures in mice.The Rho / Rho kinase signaling pathway is a key pathway that regulates the change in plasticity of dendritic spines.This study intends to use Fasudil,an inhibitor of the key protein Rock2 in the Rho / Rho kinase signaling pathway,to observe its effect on dendritic processes in epilepsy mice.Whether spines and learning and memory ability can relief,provide new treatments for learning and memory ability impairment caused by epilepsy.Objective:Observe the changes of Rho/Rho kinase signaling pathway,dendritic spines,neurons and cognitive function after epilepsy,and explore the protective and therapeutic effects of fasudil on epilepsy.Methods:At first,the effects of KA and Fasudil on neuron action potential and neuron morphology were observed on primary hippocampal neurons.Then,C57BL/6 mice were taken as the experimental subjects,and the mice were divided into two groups: Pre-Fasudil group and Post-Fasudil group.The changes of phosphorylation levels of key proteins in the Rho/Rho kinase signaling pathway were detected by western-blot in C57 mice.Active Rho Kit detects changes in Active RhoA;The density of dendritic spines in mice was observed by golgi-cox method.FJB staining was used to observe neuronal death in mice.Morris water maze,open field and hanging tail experiment were used to detect learning and memory function and anxiety and depression.Results:The frequency of action potential of hippocampal neurons increased,the length of neuronal protuberance decreased,and the number of dendritic spines decreased after treated with KA,which could be reversed by Fasudil.RhoA was activated in a short period of time after KA induced seizures in C57 mice,and phosphorylation levels of Rock2,LIMK and Cofilin were significantly changed,with the most significant change in 1h,and no significant change in protein after 1 week.The number of neuron death increased obviously.The number of dendritic spines in mice decreased irreversibly.Fasudil was given protection in advance,which could significantly reverse the change of phosphorylation level of the Rho/Rho kinase signaling pathway caused by epilepsy,protect mouse neurons,and reverse the irreversible damage of dendritic spines caused by epilepsy.After KA induced epilepsy,Fasudil was given for treatment,and there was no significant change in the key protein of Rho/Rho kinase signal in the hippocampus.The number of FJB positive cells decreased significantly,and the death of neurons was significantly improved.The irreversible damage of dendritic spines in hippocampal neurons induced by epilepsy was also improved.4 weeks after KA induced epilepsy,the behavioral experiment results showed that the learning and memory ability of KA induced epilepsy mice decreased and anxiety and depression increased.Fasudil significantly improved learning and memory impairment and anxiety and depression.Conclusion:Fasudil has protective and therapeutic effects on epilepsy in mice,and has therapeutic effects on irreversible injury of dendritic spines caused by epilepsy,learning and memory dysfunction caused by epilepsy and anxiety and depression,suggesting that Fasudil can be a new therapeutic drug for treating epileptic cognitive impairment.