| Background Seizures can cause secondary brain injury,and hippocampus structure is easy to injury.Bax is promoting apoptosis proteins,Bcl-2is inhibiting apoptosis proteins,and the two are the index of brain injury in cell apoptosis.Epidemiological and clinical studies both have identified the long-term seizures in development stage can damage learning,memory and cognitive function.At present medication is the mainly treatment of epilepsy.lt was generally believed that traditional AEDs on the influence of cognitive function is larger,but for the new AEDs,still know less the influence of cognitive function.LTG is a new type of antiepileptic drugs.the protection mechanism to the damage of the seizure and the influence of cognitive function in patients with the study of the scholars in recent years has been the hot issues.Objective Through observe the epileptic seizures and the changes of learning and memory of epileptic rat models intervented with LTG.and detect the expression of Bax and Bcl-2protein in the hippocampus.to analyze the behavior,learning and memery and the protect on of LTG whether related with Bcl-2,Bax in rats with recurrent epilepsy,and analyze the behavior, learning and memery and the expression of Bax and Bcl-2protein whether related with the dose of LTG,to search the base for LTG dose selection in clinical application.Methods One hundred six-weeks old healthy SD rats were randomly divided into the normal control group,model group,LTG low dose group,LTG medium dose group, and LTG high dose group,20cases in eath group.All the rats except those in normal group were intraperitoneal injection35mg/kg of PTZ per day,and the behavior change was observed for30min.Normal control group were intraperitoneal injection the same times.If continuously the sum above degree3add up to3times and it would be called successful kindle,and dose of normal control with the same method until the kindling criterion was reached(About2weeks).According to the largest recommended dose of LTG(600mg/d) of1/30,1/20,1/10into low, medium and high dose group.The rats in normal control group and the model group were given the same normal control instead. LTG and NS made up with the concentration.The rats was given the medicine volume for lmL/100g,once a day the rats except those in normal control group and PTZ group were intragastric administration dose of the above2weeks.From the31st,the Morris water maze was performed in models for5d.The time each group of rats which were tested in the Morris daily.Then start memory and judgment which were performed at the fifth day.After the Morris experiment,the pathology of the hippocampal were observed by HE,the expressions of Bax and Bcl-2Protein were determined by immunohistochemistry method.The data were analyzed by SPSS17.0,which were expressed as mean±SD,and the number of multiple pairwise comparisons were compared with One-Way ANOVA,and P<0.05was considered as statistical significance.Results1. The rats in normal control group were not caused seizures;In model group,the epileptic group induced by PTZ had seizure, latency of seizures was the shortest (1.70±0.79) min, and the duration of seizures was the longest (6.38±2.20) min.the3medicine administration groups had seizure too,but the level of seizure were decreased.Among them.in LTG high dose group, latency of seizures was the longest(6.81±1.86) min,and duration of seizures was the shortest(2.16±1.32) min.The difference between the normal control group and the model group in latency and duration of seizures had the statistically meaning(P<0.05).Compared with the PTZ group,the latentperiod of seizure were prolonged and the level of seizure were decreased in the group LTG drug positive group,and the difference had the statistically meaning(P<0.05).2. Escape incubation period of normal control group,model group LTG Low dose group,LTG medium dose group and LTG high dose group in the first day were respectively (25.15±1.39) s,(60.08±1.17) s,(54.50±0.93) s,(37.93±1.23) s,(36.38±0.83) s.Escape incubation period of every group in the second day were respectively (20.58±0.86) s,(61.22±1.64) s,(58.59±0.91) s,(30.70±0.95) s,(30.63±0.95) s. Escape incubation period of every group in the third day were respectively (15.45±0.61),(59.59±0.63) s,(57.31±1.06) s,(30.47±0.57) s,(25.16±1.71) s.Escape incubation period of every group in the fourth day were respectively (12.64±0.86) s,(55.62±0.40) s,(56.57±1.61) s,(27.69±0.82) s,(20.59±0.95) s.Compared with the normal control group, escape incubation period in the model group was significantly longer;Compared with the normal control group,escape incubation period of LTG Low dose group,LTG medium dose group and LTG high dose group were significantly shorter.Space exploration experiment result showed:the time staying in the platform quadrant in the model group was (26.01±2.21) s,and that in the normal control group is(48.77±3.40) s,the difference between the model group and the normal control group were significant;The rats time of staying within a platform quadrant in the time of120s,the time staying in the platform quadrant in LTG Low dose group,LTG medium dose group and LTG high dose group are (27.85±1.05) s,(37.96±0.97) s,(39.82±1.34) s.Compared with the model group,in LTG Low dose group,LTG medium dose group and LTG high dose group significantly increased.3. The results of HE staining in hippocampus indicated that hippocampal CA1pyramidal cells in the small area,CA3area and DG areas pyramidal cells in model group,LTG Low dose group,medium dose group and high dose group were arranged in loose granular cell disorder,The hippocampus cells swelling.cell borderline unclear, the nucleus and cytoplasmic hyperchromatic,the chromatin edge set into large cubes, cells degenerate vacuole-like denaturation,cell membrane retracted,and part of cell could see the apoptotic body and in model group were more severe than LTG Low dose group,medium dose group and high dose group,the necrosis of the points cells could be found. In normal control group,small pyramidal cells in Hippocampal CA1area,pyramidal cells in CA3and the granulosa cells in DG were arranged in neatly. Cells in the hippocampus was complete,borderline clear,the nucleus was round,oval.the chromatin in nuclear widely distributed,nucleoli was clear,only a little nerve cells degenerate vacuole-like denaturation,the chromatin edge set,the size increases and cell swelling assumed the circular.4. The expressions of Bax protein and Bcl-2protein in normal control group were low,and the expressions of Bax and Bcl-2protein for positive were respectively (0.16±0.39),(0.26±0.03).the expression of Bax and Bcl-2protein for positive in the model group were respectively (0.35±0.05,(0.27±0.03).Compared with the model group,the number of the positive neurons of Bcl-2protein in the LTG drug positive group increased,the number of the positive neurons of Bax protein decreased; the expression of Bax in LTG Low dose group,LTG medium dose group and LTG high dose group respectively were (0.29±0.45),(0.24±0.04),(0.20±0.05).the expression of Bcl-2in LTG Low dose group,LTG medium dose group and LTG high dose group were respectively (0.33±0.28),(0.36±0.04),(0.38±0.02).Compared with the model group,LTG Low dose group,LTG medium dose group and LTG high dose group the difference were statistically significant(P<0.05);LTG Low dose group,medium dose group and high dose group the differences of the number of positive neurons in LTG low dose group,LTG medium dose group and LTG high dose group were statistically significant (P<0.05).Conclusions1. In seizures,the changes of behavior and learning and memery are probably related to the expressions of Bcl-2/Bax in the hippocampus in epileptic rats.2. LTG may increase the expression of Bcl-2protein and decrease the expression of Bax protein of the hippocampal neurons.LTG can protect hippocampal neurons in the epileptic rats induced by PTZ and decrease the apoptosis of the hippocampal neurons after the seizure attacks.3. In certain dose rang,there did not appear adverse reaction,and with the dose of LTG increasing,the effect of LTG against apoptosis of hippocampal neurons of epileptic rats induced by PTZ enhances. |
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