目的 探讨癫痫伴抑郁模型大鼠海马突触相关蛋白突触素(synaptophysin,SYN)、突触后致密蛋白 95(postsynaptic density protein 95,PSD95)及生长相关蛋白 43(growth-associated protein 43,GAP43)的表达情况.方法 选取3月龄雌性清洁级SD大鼠进行实验,采用氯化锂-匹罗卡品建立癫痫大鼠模型,造模成功的癫痫模型大鼠根据是否伴发抑郁分为癫痫伴抑郁组(10只)和癫痫组(10只),另取体质量相匹配的10只大鼠作为抑郁组,10只作为正常组,其中抑郁组大鼠采用慢性不可预见性温和应激刺激结合孤养法建立抑郁模型.采用体质量、糖水偏爱实验和旷场实验评价大鼠的抑郁行为,免疫组织化学染色及Western blot检测大鼠海马组织SYN、PSD95、GAP43蛋白的表达.采用SPSS 17.0软件进行数据统计分析,行为学结果使用重复测量方差分析,蛋白表达数据多组间比较采用单因素方差分析,进一步两两比较采用LSD检验.结果 4组大鼠体质量比较,时间和组别的交互作用显著(F=7.33,P＜0.01),在第8天和第29天,癫痫伴抑郁组与抑郁组大鼠体质量均低于癫痫组(均P＜0.05),癫痫伴抑郁组大鼠体质量在第29天较第1天低(P＜0.05);4组大鼠糖水偏爱率比较,时间和组别的交互作用显著(F=2.67,P＜0.05),癫痫伴抑郁组大鼠在第15和29天糖水偏爱率较第1天低(均P＜0.05);旷场实验结果显示,垂直站立次数(F=2.74)与水平运动格子数(F=1.76)的交互效应作用均不显著(均P＞0.05),但时间主效应和组别主效应均显著(垂直站立次数:F时间=4.35,P＜0.05,F组间=25.64,P＜0.01;水平运动格子数:F时间=12.75,P＜0.01,F组间=21.37,P＜0.01).免疫组化结果显示,4组大鼠突触蛋白SYN、PSD95和GAP43阳性表达细胞数比较均差异有统计学意义(F=93.85,58.66,98.84,均P＜0.05).癫痫组 SYN[(11.73±4.30)个]、PSD95[(24.47± 7.58)个]、GAP43[(9.40±3.50)个]阳性细胞表达数均低于正常组[(51.00±15.39)个、(55.60± 13.17)个、(29.53±4.05)个](均 P＜0.05).癫痫伴抑郁组 SYN[(5.80±3.53)个]、PSD95[(12.87± 4.03)个]、GAP43[(5.33±3.50)个]阳性表达细胞数均低于抑郁组[(11.33±3.22)个、(48.13± 12.69)个、(15.47±5.21)个](均 P＜0.05).Western blot 结果显示,4 组大鼠突触蛋白 SYN、PSD95、GAP43的表达均差异有统计学意义(F=13.19,9.38,16.80,均P＜0.05).癫痫组的SYN、PSD95和GAP43表达水平均低于正常组(均P＜0.05).癫痫伴抑郁组SYN、PSD95和GAP43表达水平均低于癫痫组和抑郁组(均P＜0.05).结论 癫痫伴抑郁大鼠海马SYN、PSD95、GAP43蛋白的低表达可能与其发病机制有关.
Expression of synapse-related proteins in hippocampus of rats with epilepsy accompanied by depression
Objective To investigate the expression of hippocampal synapse-related proteins inclu-ding synaptophysin(SYN),postsynaptic density protein 95(PSD95)and growth-associated protein 43(GAP43)in rats with epilepsy accompanied by depression.Methods The 3-month-old female clean grade SD rats were selected for the experiment.Lithium chloride pilocarpine was used to establish an epileptic rat model.Rats with successful epilepsy models were divided into epileptic depressive group(EWD group)and epileptic group with 10 in each group based on whether they were accompanied by depression.Furthermore,ten rats with matched body mass were taken as the depressive group and 10 were taken as control group.As for the depressive group rats,chronic unpredictable mild stress stimulation combined with orphanage was a-dopted to establish a model of depression.The depressive behaviors of rats were evaluated by body mass,su-crose preference test and open field test.Immunohistochemical staining and Western blot were used to detect the expression of SYN,PSD95 and GAP43 proteins in rat hippocampal tissue.SPSS 17.0 software was used for data statistical analysis,repeated measurement ANOVA was used for behavioral results,one-way ANOVA was used for inter group comparison of protein expression data,and LSD test was used for further pairwise comparison.Results As for the body mass,there was significant interaction effect between the time and group among the 4 groups(F=7.33,P＜0.01).On the 8th day and the 29th day,the body weight of rats in the EWD group and the depressive group were lower than those in the epilepsy group(all P＜0.05).The body weight of rats in the EWD group on the 29th day was lower than that on the first day(P＜0.05).As for the sucrose preference rates,there was significant interaction effect between the time and group among the 4 groups(F=2.67,P＜0.05).The sucrose preference rate of EWD group on the 15th and 29th day were lower than that on the first day(both P＜0.05).The results of the open field test showed that the interaction effects of the number of vertical standing times(F=2.74)and the number of horizontal movement lattices(F=1.76)both were not significant(both P＞0.05),but both the time effect and group effect were signifi-cant(vertical standing times:Ftime=4.35,P＜0.05,Fgroup=25.64,P＜0.01;horizontal movement lattices:Ftime=12.75,P＜0.01,Fgroup=21.37,P＜0.01).The immunohistochemical results showed that there was a statistically significant difference in the number of positive cells expressing synaptic proteins SYN,PSD95 and GAP43 among the four groups of rats(F=93.85,58.66,98.84,all P＜0.05).The numbers of positive cells of SYN(11.73±4.30),PSD95(24.47±7.58)and GAP43(9.40±3.50)in the epilepsy group were lower than those in the control group((51.00±15.39),(55.60±13.17)and(29.53±4.05))(all P＜0.05).The numbers of positive cells of SYN(5.80±3.53),PSD95(12.87±4.03)and GAP43(5.33± 3.50)in the EWD group were lower than those in the depressive group((11.33±3.22),(48.13±12.69)and(15.47±5.21))(all P＜0.05).Western blot results showed that there were statistically significant differences in the expression of synaptic proteins SYN,PSD95 and GAP43 among the four groups of rats(F=13.19,9.38,16.80,all P＜0.05).The expression levels of SYN,PSD95 and GAP43 in the epilepsy group were lower than those in the control group(all P＜0.05).The expression levels of SYN,PSD95 and GAP43 in the EWD group were lower than those in the epilepsy group and the depressive group(all P＜0.05).Conclusion The low expression of SYN,PSD95 and GAP43 proteins in the hippocampus of rats with epilepsy accompanied by depression may be related to their pathogenesis.
NETL
NSTL
万方数据
维普
