microRNA-34b及caspase-9在托吡酯治疗颞叶癫痫大鼠海马组织中的表达及其意义

Expression and significance of microRNA-34b and caspase-9 in the hippocampus of rats with temporal lobe epilepsy treated with topiramat

[目的]探讨microRNA-34b及caspase-9在托吡酯(TPM)治疗颞叶癫痫大鼠海马组织中的表达及其意义.[方法]取雄性SD大鼠随机分为正常对照组、模型对照组及TPM低、高剂量组,腹腔注射给予氯化锂-匹罗卡品诱发大鼠癫痫发作,正常对照组和模型对照组灌胃给予生理盐水,TPM低、高剂量组分别灌胃给予TPM 40,80 mg/kg.取大脑及海马组织,行HE染色观察病理改变;釆用基因芯片分析法和实时定量RT-PCR法观察各组大鼠海马组织中microRNA-34b的表达情况;利用Western-blot蛋白免疫印迹法及免疫组织化学染色法检测各组大鼠海马组织中caspase-9水平.[结果]与正常对照组比较,模型对照组microRNA-34b及caspase-9的表达水平均显著升高(P<0.01);与模型对照组比较,TPM低、高剂量组microRNA-34b及caspase-9的表达水平均显著降低(P<0.05,P<0.01),且TPM高剂量组降低更为明显(P<0.01).[结论] TPM可下调癫痫大鼠海马microRNA-34b及caspase-9表达.

OBJECTIVE To explore the expression and significance of microRNA-34b and caspase-9 in the hippocampus of rats with temporal lobe epilepsy treated with topiramate(TPM). METHODS Male SD rats were randomly divided into normal control group, model control group and low and high dose TPM group. Lithium chloride-pilocarpine was given to induce epilepsy by intraperitoneal injection in rats. The rats in the normal and model control group were given saline by gavage, while the rats in the low and high dose TPM group were given TPM 40 and 80 mg/kg by gavage respectively. The cerebrum and the hippocampus were taken out, and pathological changes were observed by HE staining. Microarray analysis and real time quantitative RT-PCR were used to observe the expression of microRNA-34b in the hippocampus of each group. Western blot and immunohistochemical staining were used to detect the level of caspase-9 in the hippocampus of rats in each group. RESULTS As compared with the normal control group, the expressions of microRNA-34b and caspase-9 protein in the model control group were significantly increased(P<0.01). The expressions of microRNA-34b and caspase-9 in the low and high dose TPM groups were significantly reduced as compared with the model control group(P<0.05,P<0.01), in addition, the high dose TPM group decreased more significantly(P<0.01). CONCLUSION TPM could down-regulate the expressions of hippocampal microRNA-34b and caspase-9 in epileptic rats.