The Mechanism Of Reelin Involved In Actin Regulation In Oxidative Stress Neurons And Its Role In Electroacupuncture

Objective:At present,electroacupuncture therapy is widely used in the treatment of stroke patients,and achieved good curative effect.But the mechanism of electroacupuncture is not very clear.Reelin is an extracellular matrix serine protease.Previous studies have shown that the expression of Reelin protein increases after cerebral ischemia reperfusion injury in adult rats,and promotes the growth of dendritic cells in cortical ischemic region,but the mechanism is not clear.Actin cytoskeleton plays a key role in the growth and development of dendritic and dendritic spines.Then,can electric acupuncture promote the expression of Reelin protein?If EA can promote the expression of Reelin protein,is the further mechanism involved in the regulation of actin cytoskeleton by Reelin protein?This article focuses on the effect of Electroacupuncture on the expression of Reelin protein after cerebral ischemia reperfusion injury in rats and the mechanism of Reelin protein involved in regulating the actin cytoskeleton after oxidative stress.Methods:Rats were randomly divided into four groups:sham-MCAO,MCAO,MCAO+sham-EAIschmia-Sham EA group and MCAO+EA group.At 30 minutes before ischemia-reperfusion model?MCAO?and reperfusion,“philtrum-philtra“?DU26?and"pulp"?RN24?were selected for electroacupuncture treatment.Tissue protein was extracted seven days after MCAO.The expression of Reelin protein in each experimental group was detected by Western blotting and the expression of activated RhoA was detected by Rho Pull-Down assay and immunoblotting.The rats were randomly divided into four groups:sham-MCAO,MCAO,MCAO+sham-EA and MCAO+EAgroups.Using water maze,the learning and memory abilities of rats were measured.Cultured primary neurons of cerebral cortex in the neonatal rats within 24 hours were treated with different concentrations of H₂O₂ and Recombinant M ouse Reelin,and the cells were extracted after 24h.The expression levels of activated RhoA and phosphorylated cofilin were detected by Rho Pull-Down assay and Western blotting.The changes of F-actin and dendritic length and branch in cortical neurons after oxidative stress and Recombinant Mouse Reelin treatment were observed by immunofluorescence.Results:1.Compared with the control group,the learning and memory ability of MCAO group decreased significantly.However,compared with MCAO+shamMCAO,the learning and memory ability of MCAO+EA group was enganced after EA treatment.Electroacupuncture teatment can improve the learning and memory ability of MCAO rats.2.After cerebral ischemia-reperfusion injury,the expression of Reelin protein and the activation level of RhoA protein in cerebral cortex of MCAO group were increased.Giving electroacupuncture treatment,the expression of Reelin protein in the cerebral cortex was increased and the expression of activated RhoA protein was decreased.3.Oxidative stress stimulation can increase the level of RhoA-GTP and decrease the cofilin phosphorylation in primary cortical neurons.The exogenous recombinant protein—Reelin can induce a decrease in RhoA-GTP protein level and elevated levels of cofilin phosphorylation after oxidative stress injury.4.Oxidative stress stimulation can lead to decrease dendritic length and number of dendritic branches of the cerebral cortical neurons and decrease expression of F-actin.The Reelin can stimulate the growth of dendritic in cortical neurons and the polymerization of actin after oxidative stress injury.5.After oxidative stress stimulation,Wortmannin—a PI3K inhibitor can inhibit the decrease of RhoA-GTP level and the increase of cofilin phosphorylation level induced by Reelin protein.Conclusion:1.After cerebral ischemia-reperfusion injury,electroacupuncture treatment can increase the expression of Reelin and decrease the level of activated RhoA.2.After oxidative stress stimulation,Reelin activates PI3K,which inhibits the activation of RhoA on the one hand.On the other hand,it stimulates cofilin phosphorylation and promotes the polymerization of actin.