The Research Of Expression And Significance Of Neuropeptides Ghrelin And Nesfatin-1 In Kainic Acid Kindling Rats

Objective The kainic acid was injected to the amygdala of the rats and epilepsy animal model was established, we observed the expression level of neuropeptides. Ghrelin and Nesfatin-1 between the untreated and treated groups in these rats and approached the basic expression purpose, to set up the simpler, sensitive way on epilepsy diagnose and therapy assessment and to systematically study their role in nosogenesis of Epilepsy.Methods 96 male Sprague-Dawley rats were divided into NS groups and VPA groups random after seizured. Each group was divided into seven subgroups 3h, 6h, 12h, 24h, 3d, 7d and 14d, while set up blank group (without NS or VPA) and sham operation group (injected NS instead of kainic acid).Every group has 6 rats. By stereotactic operation, the kainic acid 1.0μl was injected to the amygdala of the rats of NS groups and VPA groups. We try to observe the changes of the behavior, the EEG in epileptic rats. The morphology of rats` sample was detected with Hematoxylin and Eosin (HE) coloration, Nissl coloration. The RIA, ELISA and immunohistochemistry were used to detect the expression levels of Ghrelin and Nesfatin-1 in the serum and hypothalamus of kainic acid kindling rats.Results 1. By stereotactic operation, the kainic acid was injected to the amygdala of 84 rats` brain and the epilepsy animal model was established. As result, 82 rats was induced out epilepsy, the success rate was 97.62%. By HE staining and Nissl staining induced morphological changes of neurons in epileptic rats: acute onset of irregular arrangement of neurons, astrocytes edema, degeneration; attack resting almost total absence of neurons, glial scar formation; Onset of chronic neurons have disappeared, there neuronal phagocytosis.2. The changes of Ghrelin and Nesfatin - 1 of serum in Kainic acid kindling Rats: RIA showed that between the control group and the sham operation control group was not statistical difference (p> 0.05);Compared with the control group, the expression of Ghrelin in the VPA group and the NS group were lower (p <0.05), while the NS group decreased more significantly; the VPA group compared with the NS group at different time, Ghrelin expression level of the VPA group at the corresponding time is significantly higher (p < 0.05), but still lower than the control group. ELISA showed that between the control group and the sham operation control group was not statistical difference (p > 0.05);Compared with the control group, the expression of Nesfatin-1 in the VPA group and the NS group were increased (p < 0.05), while the NS group increased more significantly; the VPA group compared with the NS group at different time, Nesfatin– 1 expression level of the VPA group at the corresponding time is significantly lower (p < 0.05), but still higher than the control group .3. The changes of Ghrelin and Nesfatin - 1 of hypothalamus in Kainic acid kindling Rats: immunohistochemistry results show that between the control group and the sham operation control group was not statistical difference (p> 0.05);Compared with the control group, the expression of Ghrelin in the VPA group and the NS group were lower (p <0.05), the VPA group compared with the NS group at different time, Ghrelin expression level of the VPA group at the corresponding time is significantly higher, (p < 0.05), especially the 24h, 7d and 14d groups were statistically significant(p < 0.05), but still lower than the control group. The expression of Nesfatin-1 in the VPA group and the NS group were increased (p < 0.05), the VPA group compared with the NS group at different time, Nesfatin-1 expression level of the VPA group at the corresponding time is significantly lower (p < 0.05), and whole the groups were statistically significant(p < 0.05),but still higher than the control group .Conclusions 1.Kainic acid kindling model with amygdala injection has the advantages in reduced the cost of experiments.2. Desacylated-Ghrlin and Nesfatin-1 in the serum could reflect the neuropeptides variance level in hypothalamus.3. The level of Ghrelin and Nesfatin-1 between the untreated and treated groups in the kainic acid kindling rats has Significant changes.The exactitude detection of expression level of desacylated-Ghrlin and Nesfatin-1 in the serum could be the simpler, sensitive and noninvasive way on epilepsy diagnose and therapy assessment.