| Objective: To investigate the expression changes of adenosine A1 receptor and ENT1 in brain tissue after inhibitting p38 MAPK signaling pathway in epileptic model rats.Methods: The healthy male SD rats were randomly divided into four groups.There were normal control group(n = 5),epilepsy group(0h,24 h,72h,1W,2W,five time points,n = 5× 5),SB203580 group(n = 5)and DMSO group(n = 5).The model of lithium pilocarpine induced seizures in rats were set up.Latent period of epileptic seizures and the number of epileptic seizures in 1 hour were observed by Racine scoring criteria.HE staining was used to observe the histopathological changes of brain tissue in each group.The expression of adenosine A1 receptor in hippocampus and temporal cortex of rats were detected by immunohistochemistry,immunofluorescence and Western blot.Western blot was used to detect the expression of ENT1 in hippocampus of each group.Results:1.Behavioral assessment results showed: The behavioral history of epileptic seizures in the epilepsy group was 25.89 ± 1.27 min,solvent control group was 25.65 ± 1.56 min,SB203580 group was 37.53 ± 1.73 min,and the number of seizures in the epilepsy group was 7.4 ± 1.07,solvent control group was 7.0 ± 0.83,SB203580 group was 3.0 ± 0.70.There was no significant difference between the first epileptic seizure latency and the number of epileptic seizures in the epilepsy group and the solvent control group(P >0.05).Compared with the above two groups,the latency of seizure was significantly prolonged in the SB203580 group,the number of episodes of seizures decreased within 1 hour,and the difference was statistically significant(P <0.05).2.The results of HE staining showed that the hippocampus had clear structure and no pathological changes in Normal control group;The hippocampus of the epilepsy group showed a large number of neuronal apoptosis and necrosis.SB203580 group hippocampus also had some neuronal degeneration necrosis,but less severe than epilepsy group.3.Western blot to detect the expression of adenosine A1 receptor results showed:normal control group rat hippocampus and new temporal lobe cortex of the expression of adenosine A1 receptors have a basic level.Lithium chloride-pilocarpine epilepsy model in rats,after the success of the epilepsy building 24 h adenosine A1 receptors expression of the increase in the normal group,adenosine A1 receptors expression peak after72 h,compared with normal control group,the difference statistically significant(P <0.05).After the application of p38 MAPK inhibitor SB203580,the expression of adenosine A1 receptor in SB203580 group was significantly lower than that in epilepsy group(72 h),and the difference was statistically significant.(P <0.05)Immunohistochemical results showed that the number of adenosine A1 receptor positive cells in hippocampus of normal control rats was 5.33 + 1.45,epilepsy group was 25.33±3.18,SB203580 group was 12.33±1.45.Compared with the normal control group,the number of positive cells of adenosine A1 receptor in epilepsy group was significantly increased,and the staining was enhanced,the difference was statistically significant(P <0.05);Compared with the epilepsy group,the adenosine A1 receptor in the epilepsy group was significantly higher than that in the normal group.The number of positive cells was decreased,and the coloration was lighter and the difference was statistically significant(P <0.05).Immunofluorescence results showed that the adenosine A1 receptors are expressed in the cell membrane and cytoplasm of neurons.4.The results of Western blot detection of ENT1 expression showed that the expression of ENT1 in hippocampus of SB203580 group(24h)was significantly lower than that of epilepsy group(24h),and the difference was statistically significant(P <0.05).Conclusions: p38 MAPK inhibitor SB203580 can reduce the pathological damage of hippocampal neurons in rats,prolong the latency of seizure in rats and decrease the degree of seizure in rats and the expression of adenosine A1 receptors and ENT1. |
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