Effects And Mechanisms Of JNK On Oxygen-glucose Deprivation In Isolated Hippocampal Slices By Regulating ENT1

Objective:The specific inhibitor of JNK,SP600125,was used to observe the changes of Neuroexcitability and the expression of ENT1 in an in vitro hippocampal slices glucose deprivation model,so as to further explore the possible mechanism by which JNK regulates the involvement of ENT1 in vitro hippocampal glucose deprivation injury.Methods: Seven days after birth,hippocampal brain slices of SD rats were prepared.The experiments were divided into normal controlgroup,model group and inhibitor group.The normal group was not treated with oxygen glucose deprivation(OGD);the model group was treated with OGD for 30 min;The inhibitor group was treated with OGD for 30 min and added with SP600125 of10?mol/L.Wesern blot was used to detect the expression level of JNK total protein(t-JNK)?phosphorylated JNK(p-JNK)and ENT1 in vitro hippocampal slices of different groups;Whole-cell patch clamp current clamp pattern was used to detect membrane potential and action potential changes of neuronsin the CA1 region of the hippocampus in different groups,and the changes ofexcitatory postsynaptic current of neurons were observed in the voltage clamp pattern.Results:1.Monitoring membrane potential and action potential by patch clamp current clamp mode results:The nerve membrane potential of the control group was in a resting state,in the model group,the absolute value of neuronal membrane potential decreased,which was statistically significant compared with the control group(P < 0.05).In the inhibitor group,the absolute value of neuronal membrane potential increased,which was statistically significant compared with the model group(P < 0.05).The action potential(AP)frequency in the model group was incresed than in control group(P < 0.05),but decreased in the inhibitor group,which was statistically compared with the model group(P < 0.05).2.Monitoring excitatory postsynaptic current by patch clamp voltage clamp mode results:In the model group,the amplitude of excitatory postsynaptic current(eEPSC)increased significantly compared with the control group(P < 0.05);in the inhibitor group,the amplitude of excitatory postsynaptic current(eEPSC)decreased significantly compared with the model group(P < 0.05).3.Western bloting results:the expression of t-JNK,?p-JNK and ENT1 were basic expressed In the control group;in the control group and inhibitor group,the expression of t-JNK was not statistically significant compared with the model group(P > 0.05);in the model group,the expression of p-JNK was increased,which was statistically significant compared with the control group(P < 0.05);after SP600125 inhibition,the expression of p-JNK was decreased,which was statistically significant compared with the model group(P < 0.05);In the model group,the expression of ENT1 was significantly increased,which was statistically significant compared with the control group(P < 0.05);in the inhibitor group,the expression level of ENT1 was significantly lower,which was statistically significant compared with the model group(P < 0.05).Conclusion: The effects of JNK inhibitor(SP600125)on Oxygen-glucose Depriva tion invitro Hippocampal Slices can reduce the expression level of ENT1,and redu ce the neuroexcitotoxicity.