发育期大鼠癫痫持续状态后海马内HIF-1α表达对Notch信号通路的影响  

Effect of hypoxia-inducible factor-1α expression on the Notch signaling pathway in the hippocampus of developing rats after status epileptics

作　　者：张林[1] 庄亚飞 李腾腾[1] 白连琴 袁宝强[1] 

机构地区：[1]徐州医学院附属医院儿童神经与康复科,徐州221000

出　　处：《神经解剖学杂志》2016年第3期339-344,共6页Chinese Journal of Neuroanatomy

基　　金：江苏省脑病生物信息重点实验室开放课题(Jsbll201)

摘　　要：目的:探究发育期大鼠癫痫持续状态(SE)后海马内低氧诱导因子-1α(HIF-1α)表达对Notch信号通路的影响。方法:出生后21 d SD大鼠168只按随机数字表法分为对照组(NS,n=56)、模型组(SE,n=56)和干预组(2ME2,n=56)。其中SE组腹腔注射10 mg/ml戊四氮(PTZ,80 mg/kg)溶液制作癫痫持续状态模型,NS组腹腔注射等剂量生理盐水,2ME2组在癫痫持续状态模型建立后立即腹腔注射2-甲氧基雌二醇(2ME2,15 mg/kg)溶液。分别于造模成功后1、4、6、8、12 h和24 h处死大鼠取出海马组织,应用Western Blot方法检测HIF-1α、Notch-1蛋白含量;另外将各组造模成功后6 h的大鼠进行灌注并取脑,应用免疫组化染色法检测海马齿状回(DG)区HIF-1α和Notch-1的阳性细胞数。结果:HIF-1α、Notch-1蛋白在NS组均平稳表达;在SE组存在明显的表达时程和表达高峰,1 h时表达即开始增多,8 h达高峰,然后逐渐减少,且各个时间点的表达均明显高于NS组,差异有统计学意义(P<0.05);HIF-1α被特异性抑制剂2ME2干预后,HIF-1α、Notch-1的表达均较SE组明显减少,差异有统计学意义(P<0.05),其中6 h时达最低值。在此时间点,SE组HIF-1α、Notch-1的阳性细胞数均明显高于NS组和2ME2组,差异有统计学意义(P<0.05)。结论:发育期大鼠癫痫持续状态后HIF-1α可能部分参与了Notch信号通路的激活与调控。Objective： To explore the effect of hypoxia-inducible factor-1α（ HIF-1α） expression on the Notch signaling pathway in the hippocampus of developing rats after status epileptics（ SE）. Methods： According to the table of random number,a total of 168 rats at postnatal 21 days were divided into control group（ NS,n = 56）,status epileptics group（ SE,n = 56） and intervention group（ 2ME2,n = 56）. SE models in the SE group were induced by intraperitoneal injection of 10 mg / ml pentetrazol（ PTZ,80 mg / kg） and rats in the NS group were injected with the same dose of saline solution; for 2ME2 group,the rats received intraperitoneal injection of the same dose of 2-methoxyestradiol（ 2ME2,15 mg/kg） immediately after the inducement. The hippocampus tissues were harvested from the rats at 1,4,6,8,12 h and 24 h after the inducement. HIF-1α and Notch-1 protein expressions were measured by Western Blot.Besides,the rats of each group were perfused by paraformaldehyde solution at 6 h after the inducement and then the brain tissues were removed. The HIF-1α and Notch-1 positive cells in the dentate gyrus were detected with immunohistochemistry method. Results： In NS group,the protein of HIF-1α and Notch-1 expressed steady; while in SE group,their expression showed up obvious time-dependent and the peak. They all began to increase at 1 h after inducement and reached the peak level at 8 h,then they all decreased gradually. However,the expression levels of HIF-1α and Notch-1protein in the SE group were significantly higher than that in NS group at each time point,the differences were significant（ P〈0. 05）. Using 2ME2,the HIF-1α specific inhibitor,HIF-1α and Notch-1 protein expression levels were significantly decreased compared with SE group,so the differences were of statistical significance（ P〈0. 05）; especially at 6 h,the expression level fall to the lowest. At this time,HIF-1α and Notch-1 positive cells in the SE group were extremely higher than that in the NS and 2ME2 groups,the

关 键 词：癫痫持续状态 海马 NOTCH信号通路 低氧诱导因子-1Α 大鼠 

分 类 号：R742.1[医药卫生—神经病学与精神病学]