| ObjectiveTo explore the effective of antiepileptic drugs on cognition in rats to guide clinical medication. To explore the relationship of antiepileptic drugs with apoptosis of hippocamp neuron . To explore the relationship of antiepileptic drugs with apoptosis of hippocamp neuron's gene expression.Methods70 adolecence male Sprague-Dawley rats ,weighing(200±20)g, were divided into seven groups randomly : normal control group (NS group), status epileptics group (Pentylenetetrazole, PTZ group), carbamazepine (CBZ) group, valproate sodium (VPA) group, phenytoin(PHT) group, topiramate (TPM)group, lamotrigine(LTG) group.Besides NS group, they were kindling by PTZ,then they were treated by antiepileptic drugs (AEDS). Three weeks later,seven groups'cognition, reaction state,behavior change and total failure rate were tested by Morris water maze. Then we check if the AEDs result in hippocamp neuron apoptosis by three methods:first,we use terminal deoxynucleo tidyl transferase (TdT) dUTP nick end labeling(TUNEL) staining method detect DNA fragment.we observe apoptosis hippocamp neuron by light microscope,we select 5 high times(1×400) field of vision and count positive and negative cell number,then we calculate the average value of positive cell rate [positive cell expression percentage =positive cell number/positive cell number + negative cell number]×100%].Second,we use flow cytometry (FCM) analyze whether AEDs cause Bcl-2 and Bax expression different in hippocamp neurons.Bcl-2 order: SC-7382, Mouse anti-Bcl-2(C-2) cloned strain,Lot:a 1805;Bax order:SC-7480,Mouse anti-Bax(B-9) cell line,Lot:l 1004.Third,we use semi-quantitative reverse transcription p olymerase chain reaction (sqRT-PCR) technology analyze the expression of bcl-2 and bax by Gel-Pro Analyzer Version 3.0. Statistical Analysis: we do statistical analysis by SPSS 11.0,data=average±standard deviation ( x±s) .we use X2 test, ANOVA(F test),q test(New-man-Keuls),χ2 rate analysis check.α=0.05,P<0.05,was consider the difference is significant.Results1 The result of Cognition: The reaction state of TPM and PHT groups are lower than other groups,some appear dull and slow. The total failure rate of NS,PHT and CBZ group is higher than PTZ,VPA and LTG groups in 4 days(P<0.05);The total failure rate of TPM group is higher than other 6 groups in 4 days(P<0.01). There is 1(1/10) rat of PTZ and TPM group and there are 2(2/10) rats of LTG group died in course. The enhancement of performance record of PTZ group, CBZ group,VPA group,LTG group has been elevated quickly(P<0.05,P<0.01),and is better than NS group. The enhancement of performance record of PHT group has been elevated slowly(P<0.05),but no difference compare with NS group. The enhancement of performance record of TPM group has been elevated slowly(P>0.05),and is worse than NS group. The time of TPM group is prolonged than other groups in searching for platform quadrant (P<0.01). The time of TPM group is shorter than CBZ group(P<0.05)and LTG group(P<0.01)in stay at platform quadrant .No difference in other groups .The average of TPM group is longer than other groups in every time (P<0.05). The time of TPM group is longer than LTG groups in 1st day(P<0.05). The time of TPM group is longer than VPA and LTG groups in 2nd day (P<0.05). The time of TPM group is longer than PTZ and LTG groups in 3rd day (P<0.05). The time of TPM group is longer than CBZ, VPA and LTG groups in 4th day (P<0.05). The total time of LTG group is shorter than other 6 groups in six times (P<0.05, P<0.01). The total time of TPM group is longer than other groups in six times (P<0.01). The total time of PHT group is longer than other 5 groups in six times (P<0.05, P<0.01). The total time of TPM group is longer than CBZ, VPA, PTZ group (P<0.05) and LTG group (P<0.01) in 4 days. The time of TPM group is prolonged than other groups in searching for platform quadrant (P<0.01). The time of TPM group is shorter than CBZ group (P<0.01) and LTG group (P<0.01) in stay at platform quadrant.2 The result of TUNEL: In hippocamp CA1,TUNEL positive cell rate as follows:NS group is 2.79%,PTZ group is 3.72%,CBZ group is 5.70%,VPA group is 3.56%,no significant difference between 4 groups(X2=3.59, P>0.05);PHT group is 33.68%,TPM group is 35.79%,LTG group is 34.02%,there are no significant difference between 3 gruops (X2=0.32, P>0.05). there are significant difference between NS,PTZ,CBZ and VPA groups compare with PHT,TPM and LTG groups (X2=329, P<0.005). In hippocamp CA3,TUNEL positive cell rate as follows:NS group is 2.31%,PTZgroup is 2.97%,CBZ group is 3.66%,VPA group is 2.61%,no significant difference between 4 groups(X2=7.70, P>0.05);PHT group is 25.2%,TPM group is 30.16%,LTG group is 30.15%,there are no significant difference between 3 gruops (X2=2.03, P>0.05). there are significant difference between NS,PTZ,CBZ and VPA groups compare with PHT,TPM and LTG groups (X2=253.43, P<0.005). In hippocamp dentate gyrus,TUNEL positive cell rate as follows:NS group is 3.41%,PTZgroup is 4.01%,CBZ group is 4.42%,VPA group is 2.52%,no significant difference between 4 groups(X2=1.683, P>0.05);PHT group is 30.58%,TPM group is 32.98%,LTG group is 29.35%,there are no significant difference between 3 gruops (X2=0.892, P>0.05). there are significant difference between NS,PTZ,CBZ and VPA groups compare with PHT,TPM and LTG groups (X2=287.092, P<0.005).3 The result of flow cytometry: The comparion of Bcl-2 expression has significant difference between follow couples:NS vs PHT (q=4.887, P<0.05),NS vs TPM(q=3.858,P<0.05), NS vs LTG (q=3.734, P<0.05);PHT vs PTZ(q=3.399,P<0.05); there are no significant difference between other gruops (P>0.05).The comparion of Bcl-2 linear expression has significant difference between follow couples:NS vs PHT(q=4.887,P<0.05), NS vs CBZ (q=3.650,P<0.05), NS vs LTG (q=3.724,P<0.05);PHT vs PTZ (q=3.146, P<0.05); there are no significant difference between other gruops (P>0.05). The comparion of Bcl-2 FI expression has significant difference between follow couples:PHT vs PTZ (q=2.542,P<0.05),PHT vs VPA(q=2.657,P<0.05); there are no significant difference between other gruops (P>0.05). The comparion of Bax expression has significant difference between follow couples:TPM vs PTZ (q=3.241,P<0.05),TPM vs CBZ(q=3.477,P<0.05); there are no significant difference between other gruops (P>0.05). There are no significant difference between Bax linear expression (F=0.471,P>0.05). There are no significant difference between Bax FI expression (F=0.516,P>0.05). The comparion of Bcl-2/Bax expression has significant difference between follow couples:NS vs PHT(q=4.464,P<0.05), NS vs CBZ(q=5.625,P<0.05), NS vs LTG(q=3.701,P<0.05);PHT vs PTZ(q=3.395,P<0.05); PHT vs TPM (q=4.259,P<0.05); CBZ vs TPM(q=4.259,P<0.05);there are no significant difference between other gruops (P>0.05). The comparion of Bcl-2(FI)/ Bax(FI) expression has significant difference between follow couples: PHT vs PTZ(q=3.465,P<0.05); PHT vs PTZ(q=3.465,P<0.05); PHT vs VPA (q=5.750, P<0.05);PHT vs TPM(q=5.088,P<0.05); PHT vs LTG (q=3.374,P<0.05); there are no significant difference between other gruops (P>0.05).4 The result of RT-PCR: The bcl-2 expression and bcl-2/β-actin no significant difference (F=0.312,F=0.199,P>0.05);The bax expression no significant difference (F=0.792,P>0.05). The relative value of bax/β-actin have signifcante difference as follow couples(F=1.943,P<0.05),NS vs PHT组(q=3.469, P<0.05),NS vs VPA(q=3.420,P<0.05), there are no significant difference between other couples(P>0.05). There are no significant difference between bcl-2/bax and bax/ bcl-2 (F=0.652,F=1.756,P>0.05) in all groups.Conclusion1 Anti-epileptic drug could affect cognition of PTZ induce epileptic seizure rats, TPM damage function of study,memory,and judgement of rats,PHT damage function of study and judgement of rats;VPA,CBZ and LTG could elevate cognition and reacting state of rats.2 The TUNEL positive rate of hippocamps is high in PHT group,TPM group and LTG group. The TUNEL positive rate of hippocamps is low in NS group,PTZ group, CBZ group and LTG group.3 Epilepsy of PTZ induce epileptic seizure rats treated by anti-epileptic drug(PHT,CBZ,LTG) could increase Bcl-2 expression,the difference of Bcl-2 and Bcl-2/Bax is significant compare with NS group(P<0.05); The difference of Bcl-2 and Bcl-2/Bax is no-significant compare with NS group in other groups(P>0.05). Epilepsy of PTZ induce epileptic seizure rats treated by anti-epileptic drug could down regulation Bax expression,the difference of Bax is no-significant compare with NS group(P>0.05).4 Epilepsy of PTZ induce epileptic seizure rats treated by anti-epileptic drugs could increase bcl-2 expression and bcl-2/β-actin,the difference of bcl-2 and bcl-2/β-actin is no-significant compare with NS group(P>0.05); The difference of VAP group of bax expression is significant compare with NS group (P<0.05);. The difference is no-significant compare with other groups(P>0.05).The relative value of bax/β-actin is low in PHT and VPA group,the difference is significant compare with NS group(P<0.05); The difference is no-significant compare with other groups (P>0.05).The expression of bcl-2 and bax is difference in 7 groups,but most difference has no-significant(P>0.05),the reason maybe as an anti-epileptics drug couldn't induce apoptosis gene express in a big amount.
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