| Objective:(1) To observe the dynamic change of behavior and cognitive function in epilepsy rats induced by Lithium-Pilocarpine (LI-PILO); (2) To investigate the dynamic changes of H2S and CBS levels in rat hippocampus and the potential role of H2S/H2S in epileptogenesis. Methods:Male SD rats were randomly divided into four groups:normal control group(NC group), epilepsy group(EP group), EP+HA (Hydroxylamine, HA) group(HA group), EP+NaHS (Sodium hydrosulfide, NaHS) group(NaHS group). The epilepsy rat model was induced by intraperitoneally administration of LI-PILO. NaHS(exogenous donor of H2S,14μmol/kg) and HA(CBS inhibitor,12.5mg/kg) was injected intraperitoneally respectively 25 minutes before PILO administration. The ability of spacial learning and memory of epilepsy rats was tested by Morris water maze. The level of H2S in plasma was detected by sensitive sulphur electrode method. The expression of CBS mRNA, c-fos mRNA and CREB mRNA was observed by Real time PCR. The expressions ofβ-actin and CBS proteins were examined by western blotting. Results:①The success rate of epilepsy rats model with acute attack induced by Lithium-Pilocarpine is 91.7% and spontaneity seizure would attack in the model rats during chronic phase. It is an ideal kind of model used to explore the formation mechanism of the temporal epilepsy.②Compared with NC group, the values of H2S concentrations in the plasma, the expression of CBS mRNA and protein in hippocampus of EP rats were up-regulated (P< 0.05) in acute attack phase (2h and 24h), conversely, in 7d and 30d, the level of H2S/CBS were down-regulated (P<0.05).③NaHS, an exogenous H2S donor, causes the production of H2S/CBS. H2S/CBS level in NaHS group was higher than EP and HA group at 2h,4h,7d and 30d(P<0.05).④Epilepsy delitescence in NaHS group was longer than EP and HA group. However, there was no significant difference(p>0.05). The times of spontaneity attack of NaHS group was less than EP and HA group in chronic phase(P<0.05). There was no significant difference between EP group and HA group(P> 0.05).⑤Water maze test showed that the spatial learning and memory ability of EP rats decreased. In EP rats, the escape latency and searching distance to find the platform were significantly prolonged or extend (compared with NC group, P<0.05). The times of passing plateform, mean exploring time and percentage of swimming distance in original platform quadrant were decreased significantly(compared with NC group, P<0.05). NaHS can improve the ability of learning and memory, all indices of spatial learning and memory had significant differences (compared with EP and HA group, P<0.05).⑥The expression of c-fos mRNA were up-regulated in EP group, HA group, NaHS group at 2h,24h and 7d (Compared with NC group, P<0.05) and down-regulated at 30d time(P<0.05). The expression of c-fos mRNA down-regulated in NaHS group at 2h (Compared with EP group, P<0.05).⑦Compared with NC group, the expression of CREB mRNA of EP rats were up-regulated in acute period (2h and 24h, P<0.05) and down-regulated in chronic period (7d and 30d, P<0.05). The expression of CREB mRNA in NaHS group was higher than EP group and HA group at 24h,7d and 30d (P<0.05). There were no statistical significance at 7d and 30d(compared with NC group, P>0.05). Conclusion:The expression of H2S and CBS increased in acute attack phase of EP rats and decreased in chronic spontaneity of the seizure, which illustrate that H2S/CBS may participate in the attack of seizures disease. Epilepsy disease may impair the ability of learning and memory in rats. Exogenous H2S donor NaHS can up-regulate the expression level of H2S/CBS, relieve epilepsy attack and improve the learning and memory ability of EP rats. The expression of the c-fos mRNA and CREB mRNA in hippocampus of EP rats were up-regulated in acute period and down-regulated in chronic phase. Exogenous In summary, NaHS, an exogenous H2S donor, can increase the expression of H2S/CBS, which is associated with the increased expression of the c-fos mRNA and CREB mRNA in chronic phase of EP rats. |
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