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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 investigate the effect of hypoxia inducible factor-1α (HIF-1α) expression in the hippocampus on the Notch signaling pathway in seizure rats after developmental status epilepticus (SE). Methods: 168 SD rats were randomly divided into control group (NS, n = 56), model group (SE, n = 56) and intervention group (2ME2, n = 56) at 21 d after birth. SE group was intraperitoneally injected with 10 mg / ml pentazotin (PTZ, 80 mg / kg) solution to make status epilepticus model. NS group was given intraperitoneal injection of equal dose of normal saline. 2ME2 group was injected intraperitoneally immediately after establishment of status epilepticus model 2 - Methoxyestradiol (2ME2, 15 mg / kg) solution. The rats were sacrificed at 1, 4, 6, 8, 12 h and 24 h after successful model establishment, respectively. The contents of HIF-1α and Notch-1 protein were detected by Western Blot. h were perfused and the brain was harvested. The number of HIF-1α and Notch-1 positive cells in hippocampus dentate gyrus (DG) area was detected by immunohistochemistry. Results: The expression of HIF-1α and Notch-1 protein in NS group was stable. In SE group, the expression of HIF-1α and Notch-1 protein were peaked at 1 h and reached the peak at 8 h, then decreased gradually. (P <0.05). The expression of HIF-1α and Notch-1 in HIF-1α was significantly lower than that in SE group after intervention with 2ME2, a specific inhibitor of HIF-1α, the difference was statistically significant There was statistical significance (P <0.05), of which 6 h reached the lowest value. At this time point, the positive cells of HIF-1α and Notch-1 in SE group were significantly higher than those in NS group and 2ME2 group (P <0.05). CONCLUSION: HIF-1α may be partly involved in the activation and regulation of Notch signaling pathway in developing rats with status epilepticus.