摘要
目的探讨不同剂量拉莫三嗪对戊四氮所致癫痫大鼠的行为学、海马组织形态学及Caspase-3和热休克蛋白70(HSP70)表达的影响。方法选取清洁级健康6周龄雄性Sprague-Dawley大鼠100只,随机分为正常对照组、模型组和拉莫三嗪低、中、高剂量组,每组20只。正常对照组大鼠腹腔注射生理盐水(3.5 m L·kg-1),其余各组大鼠均腹腔注射戊四氮(35 mg·kg-1)并观察其行为学变化。以连续出现3次Ⅳ级及其以上发作作为点燃标准。全部点燃后,模型组和拉莫三嗪低、中、高剂量组大鼠给予戊四氮35 mg·kg-1每日1次腹腔注射维持点燃,拉莫三嗪低、中、高剂量大鼠组分别给予拉莫三嗪20、30、60 mg·kg-1·d-1灌胃,连续2周。在末次灌胃24 h内,对各组大鼠进行心脏灌注固定取海马组织,采用免疫组织化学法检测海马CA3区Caspase-3和HSP70蛋白表达。结果除正常对照组外,其余各组大鼠均有痫性发作。正常对照组大鼠海马结构完整,其他组大鼠海马神经元体积缩小,与周围组织有脱离,细胞核深染,有核分裂及核溶解,具有凋亡特征,多数细胞质呈空泡状,海马区神经元细胞排列散乱,无层次感,尼氏体减少,部分神经元丢失,细胞密度小。与正常对照组比较,模型组大鼠海马CA3区Caspase-3和HSP70阳性细胞数增多(P<0.01);拉莫三嗪低、中、高剂量组大鼠海马CA3区Caspase-3阳性细胞数均少于模型组,HSP70阳性细胞数均多于模型组(P<0.05);拉莫三嗪低、中、高剂量组大鼠海马CA3区Caspase-3阳性细胞数随着剂量的增加而减少,HSP70阳性细胞数随着剂量的增加而增加,拉莫三嗪低、中、高剂量组大鼠海马CA3区Caspase-3、HSP70阳性细胞数两两比较差异均有统计学意义(P<0.05)。与正常对照组比较,模型组大鼠海马CA3区Caspase-3及HSP70蛋白表达均增加(P<0.05)。拉莫三嗪低、中、高剂量组Caspase-3蛋白表达低于模型组(P<0.05),HSP70蛋白表达高于模型组(P<0.05)。拉莫三嗪低、中、高剂量组大鼠海马CA3区Caspase-3蛋白表达随着剂量的增加而降低,HSP70蛋白表达随着剂量的增加而增加,拉莫三嗪低、中、高剂量组大鼠海马CA3区Caspase-3、HSP70蛋白表达两两比较差异均有统计学意义(P<0.05)。结论拉莫三嗪对戊四氮所致癫痫大鼠的癫痫发作有保护作用,可以减轻大鼠癫痫发作的严重程度,其机制可能是通过降低海马组织中Caspase-3表达、增加HSP70表达来抑制癫痫过程中的神经元凋亡。
Objective To explore the effect of different dose of lamotrigine on ethology,hippocampus histomorphology,the expression of Caspase-3 and heat shock protein 70(HSP70)in hippocampus of epilepsy rats induced by pentetrazole.Methods One hundred clean healthy 6 weeks old male Sprague-Dawley rats were selected and randomly divided into normal control group,model group,low-dose lamotrigine group,medium-dose lamotrigine group and high-dose lamotrigine group,with 20 rats in each group.The rats in normal control group were intraperitoneally injected saline(3.5 mL·kg-1);the rats in the other group were intraperitoneally injected pentrolone(35 mg·kg-1)to establish the epilepsy model and then the change of ethology of rats were observed.Three times consecutive episodes(grade IV and above)were used as kindling criteria.The rats in model group,low-dose lamotrigine group,medium-dose lamotrigine group and high-dose lamotrigine group were intraperitoneally injected pentrolone(35 mg·kg-1)to sustain kindling At the same time,the rats in low-dose lamotrigine group,medium-dose lamotrigine group and high-dose lamotrigine group were given lamotrigine 20,30,60 mg·kg-1·d-1 respectively by intragastric administration for two weeks.Within 24 h after the last intragastric administration,the hippocampus specimens were taken from all rats by heart perfusion.The expression of Caspase-3 and HSP70 were detected by immunohistochemistry.Results All rats had seizures except normal control group.The hippocampus formation of rats in normal control group was normal.In other four groups,the volume of neurons in hippocampus of rats were small,and the neurons were separated from surrounding tissue;the hyperchromatic nucleus,karyokinesis and karyolysis were observed and most of kytoplasm appeared vacuolus and disorder of arrangement;the nissl bodies were diminished,some neurons were lost,and the cell density was small.The number of Caspase-3 and HSP70 positive cells in CA3 zone of hippocampus of rats in model group was significantly more than that in the normal control group(P<0.01).The number of Caspase-3 positive cells in CA3 zone of hippocampus of rats in low-dose lamotrigine group,medium-dose lamotrigine group and high-dose lamotrigine group was less than that in the model group(P<0.05),but the number of HSP70 positive cells in CA3 zone of hippocampus of rats in low-dose lamotrigine group,medium-dose lamotrigine group and high-dose lamotrigine group was more than that in the model group(P<0.05).There was statistic difference in the number of Caspase-3 and HSP70 positive cells in CA3 zone of hippocampus of rats among low-dose lamotrigine group,medium-dose lamotrigine group and high-dose lamotrigine group(P<0.05);the number of Caspase-3 positive cells in CA3 zone of hippocampus of rats decreased with the increase of the dose of lamotrigine,the number of HSP70 positive cells in CA3 zone of hippocampus of rats increased with the increase of the dose of lamotrigine.The expression of Caspase-3 and HSP70 protein in CA3 zone of hippocampus of rats in model group was significantly higher than that in the normal control group.The expression of Caspase-3 protein in CA3 zone of hippocampus of rats in low-dose lamotrigine group,medium-dose lamotrigine group and high-dose lamotrigine group was significantly lower than that in the model group(P<0.05),but the expression of HSP70 protein was significantly higher than that in the model group(P<0.05).There was statistic difference in the expression of Caspase-3 and HSP70 protein in CA3 zone of hippocampus of rats among low-dose lamotrigine group,medium-dose lamotrigine group and high-dose lamotrigine group(P<0.05);the expression of Caspase-3 protein in CA3 zone of hippocampus of rats decreased with the increase of the dose of lamotrigine,the expression of HSP70 protein in CA3 zone of hippocampus of rats increased with the increase of the dose of lamotrigine;there was statistic difference in the expression of Caspase-3 protein and HSP70 protein in CA3 zone of hippocampus of rats among low-dose lamotrigine group,medium-dose lamotrigine group and high-dose lamotrigine group(P<0.05).Conclusion Lamotrigine can protect hippocampal neurons in epileptic rats induced by pentetrazole and decrease the degree of epileptic seizure,which may be achieved through inhibiting the apoptosis of neurons by increasing the expression of HSP70 protein and decreasing the expression of Caspase-3 protein in hippocampal tissues.
作者
高静
石景鹤
王家勤
张迎辉
GAO Jing;SHI Jing-he;WANG Jia-qin;ZHANG Ying-hui(Department of Pediatrics,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Department of Pediatrics,Zhengzhou Children′s Hospital,Zhengzhou 450053,Henan Province,China;Department of Children Rehabilitation,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan Province,China)
出处
《新乡医学院学报》
CAS
2018年第4期272-276,共5页
Journal of Xinxiang Medical University
基金
河南省医学科技攻关计划项目(编号:201003077)
关键词
拉莫三嗪
癫痫
大鼠
CASPASE-3
热休克蛋白70
lamotrigine
epilepsy
rat
containing cysteine aspartic acid protease-3
heat shock protein 70
