| ObjectiveThe study establish the model of chronic temporal lobe epileptic rats induced by lithium-pilocarpine. Research the effect of new anti-epileptic drug pregabalin on the expression of bcl-2and bax in hippocampus of chronic temporal lobe epileptic rats. To explore the anti-epileptic pharmacology mechanism of pregabalin and its anti-apoptotic effect on hippocampal neurons of rats with temporal lobe epilepsy(TLE).MethodsThirty male Sprague-Dawley (SD) rats were randomlly divided into normal saline(NS) control group, model group and pregabalin treatment group (n=10) according to body weight. Establish the model of chronic temporal lobe epileptic rats induced by lithium-pilocarpine. When the rats reached seizure V grade1hour, sulfate atropine and diazepam were given in intraperitoneally to relieve seizures. The control group was injected equal NS instead of lithium chloride and pilocarpine. To observe three weeks, then the rats in pregabalin treatment group received intraperitoneal injection of pregabalin (40mg/kg) once daily for three weeks. While the control group and TLE model group were injected equal NS intrapreitoneally. The expression of bcl-2and bax in hippocampus of rats was detected by immunohistochemical technique and western blot.ResulTs1. Lithium-pilocarpine induced rats epilepsy model are established. The changes in behavior experienced three phases, which were acute, silent and chronic. The rate of successful slatus epilepticus (SE) model was85.2%and mortality was25.9%. All the surviving SE rats were induced to spontaneous epilepsy and recurrence of symptoms two weeks after SE. The rate of successful chronic model was74.1%. After the treatment of pregabalin, the seizure of rats alleviated gradually.2. Bax and Bcl-2in the hippocampus was detected in normal saline control group, model group and pregabalin treatment group respectively using immunohistochemistry technique. Bax and Bcl-2positive cells located cytoplasm and nuclear membrane, shouwing brown. The results showed:1) Bax was slightly expressed in normal saline control group rats. Compared with normal saline control group, Bax positive neurons in TLE model group increased significantly(P<0.05). Pregabalin treatment can down-regualated the expression of Bax significantly compared to TLE model group rats, but still higher than normal saline control group rats(P<0.05).2) Bcl-2was slightly expressed in normal saline control group rats. In model group the Bcl-2positive neurons in hippocampus increased significantly(P<0.05). Pregabalin treatment up-regulated the expression of bcl-2significantly compared to TLE model group rats(P<0.05).3. Bax and Bcl-2in the hippocampus was detected in normal saline control group, model group and pregabalin treatment group respectively using western blot experiments. The results showed:1) Bax was slightly expressed in normal saline control group rats. Compared with normal saline control group, the expression of Bax in the hippocampus of rats with TLE increased significantly (P<0.05). Pregabalin treatment down-regulated the expression of Bax compared to TLE model group rats, but still higher than normal saline control group rats(P<0.05).2) Bcl-2was slightly expressed in normal saline control group rats. Compared with normal saline control group, the expression of Bcl-2in the hippocampus of rats with TLE increased significantly (P<0.05). Pregabalin treatment up-regulated the expression of Bcl-2significantly compared to TLE model group rats (P<0.05).Conclusion1The chronic temporal lobe epileptic rat model induced by lithium-pilocarpine was an ideal animal model to simulate human TLE, and it could be used for the reseach.2The expression of apoptotic reglulatory genes Bax and Bcl-2increased in temporal lobe epileptic chronic stage, explain Bax participate the neuron apoptotic mechanism in the hippocampus of TLE rats, thus promote neuton cells apoptosis.3Pregabalin has the effect of alleviating the number and dgree of seizures, prompt pregabalin can cure epilepsy.4Pregabalin can down-rugulate the expression of Bax and up-regulate the expression of Bcl-2in the hippocampus of chronic temporal lobe epileptic rats. So pregabalin has the effect of inhibiting cell apoptosis and protecting neurons. |
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