| Objective: 1.To explore the changes of mental behavior and cognitive function in the latent and chronic stages of epileptic rats;2.To investigate the changes of hippocampal expression of synaptic vesicle protein 2A(SV2A),laminin β1 and integrininβ1 in epileptic rats in the latent and chronic phase of epileptic rats.Methods: 1.A total of 199 healthy adult male SD rats were randomly selected as normal control groups,and the remaining 193 rats were used to prepare epilepsy models.A lithium chloride-pirocamine acute epileptic model was prepared by using lithium chloridepirocamine combination to induce rats to enter status epilepticus.3-7 days after the model is established for the incubation period of the epilepsy rat model,that is,it is included in the incubation period,and after 14 days,the rat enters the chronic phase,and the video is continuously monitored for 1 week,and the time and level of each seizure of the rat are recorded,and the rat with spontaneous recurrent seizures is a chronic epileptic rat,that is,it is included in the chronic phase group.2.Six normal rats were selected as the normal control group,and 8 rats were selected as the latency group and chronic phase group in the incubation period and chronic phase of the model,and then behavioral experiments such as elevated cross maze,sugar water preference experiment,mine experiment,and Y maze were carried out.3.Five normal rats were selected as the normal control group,and five rats were selected in the incubation period and chronic phase of the model as the latent group and chronic phase group,and the brain tissues of each group were taken from the brains of each group for HE staining,immunohistochemistry and western blotting experiments,and the changes of SV2 A,laminin β1 and integratin β1 in the hippocampus of the three groups were detected.Results: 1.Model preparation: 193 SD rats were divided into 139 8-10-week-old adult rats and 54 3-4-week-old juvenile rats,and the lithium-piropine epilepsy model was established:(1)A total of 139 adult rats were successfully ignited,118 were successfully ignited,and 21 were unsuccessful;(2)There were 54 juvenile rats,38 were successfully ignited,and 16 rats were unsuccessfully ignited.(3)Total: 156 were successfully ignited,accounting for 80.83%,and 37 were not successfully ignited,accounting for 19.17%;Successful ignition of rat deaths: 156 rats successfully ignited,68 survived,accounting for 43.59,88 died,proportion: 56.41%: of which 71 died in the acute phase,accounting for: 80.68%,the incubation period deaths: 14,accounted for: 15.91%,and 3 died in the chronic phase,accounting for 3.41%;Mortality and survival of adult rats and juvenile rats who were successfully ignited: 73 of the 88 rats that died;Fifteen juvenile rats died.Adult rats had higher mortality rates than juvenile rats,and survival was lower than that of juvenile rats,with a P<0.05.2.Video surveillance results:Among the 68 rats that survived,36 rats did not develop spontaneous epilepsy and did not develop into rats with chronic epilepsy,accounting for 52.94%.32 rats with spontaneous epilepsy became chronic epilepsy,accounting for 47.06%;Among the 32 mice with chronic epilepsy,only 18 had grade III seizures,accounting for 56.25%,and 14 had IV-V seizures,accounting for 43.75%.Each rat with chronic epilepsy had an average of 3.69±3.32 seizures per week,and the average time of each seizure was 37.15±29.15 s.3.Behavioral results : The results of the Y maze spontaneous alternation experiment showed that there were significant differences in the accuracy rate of normal control group,latent phase group and chronic phase group(F=17.654,P=0.000),the accuracy rate of normal control group and the latent phase group(P=0.001)was significantly reduced,and the accuracy rate was significantly reduced between normal control group and chronic phase group(P=0.000),indicating that the spatial memory was impaired between normal control group and the latent period group and chronic phase group,while there was no statistical difference between the latent phase group and the chronic phase group.The results of the elevated cross maze experiment showed that the normal control group,the latent stage group and the chronic phase group entered the open arm(openarmentry,0E)(H=6.165,P=0.046),the time to enter the open arm(openarmtime,0T)(F=5.213,P=0.016),and the time to enter the closed arm(CT)(F=5.452,P=0.013),There was a significant difference in the percentage of time to enter the arm opening time OT(%)(F=6.737,P=0.006),the percentage OE(%)(F=5.688,P=0.012)of the total number of opening times,and the results were not statistically different between the normal control group and the incubation period group,and the OE(P=0.04),OT(P=0.012),OT(%)(P=0.003)and OE(%)(P=)(P=)of the normal control group compared with the chronic phase group 0.003)decreased significantly,and CT(P=0.004)increased significantly,indicating that the chronic phase group showed more obvious anxiety-like symptoms than the normal control group.Compared with the chronic stage,OT(P=0.013)and OT(%)(P=0.015)were significantly reduced in the chronic phase group,indicating that the chronic phase group had more anxiety-like symptoms than the latent stage group.The results of mine experiments showed that there were significant differences between the normal control group,the incubation period group and the chronic phase group in the number of standing times(F=4.712,P=0.022)and the number of entering the center(F=17.225,P=0.000).Compared with the incubation period,the incubation group entered the center less times(P=0.030)and stood less(P=0.001).This indicates that the incubation group showed more anxiety-like emotions than the normal group.Compared with the normal control group and the chronic phase group,the chronic phase group entered the center less often(P=0.000)and stood less often(P=0.008)than the normal control group,indicating that the chronic phase group showed more severe anxiety-like symptoms than the normal control group,and the results were not statistically significant compared with the chronic phase group in the latent phase group and the chronic phase group.The results of sugar water preference experiments showed that there were significant differences in sugar water preference rates between normal control group,latent group and chronic phase group(F=15.01,P=0.000),compared with normal control group and incubation period,the sugar water preference rate in the incubation group was significantly lower than that in normal group(P=0.048),the latent group was more depressed than the normal control group,and the sugar water preference rate in the chronic phase group was significantly lower than that in the normal control group(P=0.000)and the incubation group(P=)compared with the chronic phase group 0.002),indicating that the chronic phase group showed a more depressed state than the normal control group and the latent stage group.4.SV2 A,laminin β1 and integratin β1 were expressed in hippocampal tissues: immunohistochemical results showed that the expression of SV2 A,laminin β1 and integrinin β1 was significantly downregulated in the dentate gyrus region of the hippocampus during the latent stage and rebounded in the chronic phase.The results of western blotting showed that SV2 A,laminin β1 and integratin β1 had significant differences in the normal control group,latent phase group and chronic phase group(F=12.153,P=0.008),(F=31.495,P=0.001)and(F=5.981,P=0.037),and the subsequent pairwise comparison showed that SV2 A,laminin β1 and integratin β1 decreased significantly in the epilepsy latency period,compared with the normal control group and the chronic phase group.P<0.05 indicates that the expression of SV2 A,laminin β1 and integrin-β1 in the hippocampus decreased significantly in the latency period and gradually increased in the chronic phase.Conclusions: 1.Epilepsy rats showed different degrees of depression,anxiety and spatial memory impairment in the chronic phase and the latent period,and the chronic phase was more obvious than the latent period.2.Synaptic vesicle protein 2A,laminin β1 and integratin β1 gradually decreased in the epileptic latent period,and the expression of the three in hippocampal tissues gradually increased in the chronic phase. |
PDF Full Text Request