戊四氮诱导发育鼠癫痫持续状态后海马区HIF-1α的表达及其与神经干细胞增殖的关系  

Expression of hypoxia-inducible factor-1 alpha and correlation between HIF-1α level and proliferation of neural stem cells in the hippocampus of developing rats after pentylenetetrazol- induced status epilepticus

作　　者：周保健[1] 袁宝强[2] 

机构地区：[1]江苏省泰兴市人民医院儿科,225400 [2]徐州医学院附属医院儿科,徐州221000

出　　处：《中华神经医学杂志》2012年第4期347-352,共6页Chinese Journal of Neuromedicine

基　　金：江苏省高校自然科学基金（07KJB320120）

摘　　要：目的研究低氧诱导因子-1α（HIF-1α及nestin蛋白在戊四氮（PTZ）诱导发育期大鼠癫痫持续状态（SE）后海马区的表达变化，探讨HIF-1α与神经干细胞州SCs）增殖的关系。方法出生后7、14、21、28dSD大鼠各192只，按随机数字表法分为模型组（96只）和对照组（96只），腹腔注射10g／L戊四氮（PTZ）溶液制作SE模型，对照组注射等剂量生理盐水，造模后6h、12h、1d、2d、3d、7d采用RT．PCR、免疫组化分别检测各组大鼠海马区HIF．1dmRNA和蛋白的表达：出生后21d大鼠48只按随机数字表法分为模型组（24只）和干预组（24只），其中干预组大鼠右侧海马齿状区微量注射1μL HIF-1α反义寡聚核苷酸（ASODN），左侧注射等体积生理盐水，6h后2组大鼠均造模。造模后2d采用RT．PCR检测大鼠海马区HIF-1α mRNA的表达．造模后7d、14d免疫组化检测海马区HIF-1α、nesfin的表达。结果与对照组（未检测到）比较，各日龄大鼠SE后海马区HIF．1amRNA的表达明显升高，表达趋势一致。SE后6h时HIF-1α mRNA的表达随日龄增大而下降，差异有统计学意义（P〈0．05）。各日龄大鼠SE后各时间点海马区H1F-1α阳性细胞分布的部位随时间变化而不同；与模型组和干预组注射生理盐水侧比较，SE后2d干预组注射ASODN侧HIF-1α mRNA的表达明显降低，SE后7d、14d海马区nestin阳性细胞计数较少，差异均有统计学意义（P〈0．05）。结论发育期大鼠SE后海马区HIF-1α mRNA及其蛋白的表达增强．与日龄有关；SE后HIF-1α对NSCs增殖可能存在调控作用。Objective To explore the expressions of hypoxia-inducible factor-1 alpha （HIF-1α and nestin in the hippocampus of developing rats after pentylenetetrazol （PTZ）-induced status epilepticus （SE）, and the correlation between HIF-1α level and proliferation of neural stem cells （NSCs）. Methods A total of 768 SD rats, including rats of postnatal 7 days （P7, n=192）, rats of postnatal 14 days （P14, n=192）, rats of postnatal 21 days （P21, n=192） and rats of postnatal 28 days （P28, n=192）, were equally randomized into vehicle group （n=96） and control group （n=96）, respectively; SE rat models of the vehicle group were induced by intraperitoneal injection of 1% PTZ （80 mg/kg）; and rats of the control group were received injection of normal saline （NS）; 6 and 12 h, and 1, 2, 3 and 7 d after the inducement, the mRNA and protein expressions of HIF-1α in the hippocampus of rats in these 8 groups were examined by RT-PCR and immunohistochemistry, respectively. Another 48 21-d- old SD rats were equally randomized into vehicle I group （n=24） and intervention group （n=24）; and rats in the intervention group received injection of HIF-1αantisense oligodeoxynucleotides （ASODN） into the right hippocampus and NS into the left hippocampus; 6 h after the injection, these 48 rats were induced into SE models by intraperitoneal injection of 1% PTZ （80 mg/kg）; 2 d after the inducement, the mRNA expression of HIF-1α in the hippocampus was examined by RT-PCR; and 7 and 14 d after the inducement, the protein expressions of HIF-1α and nestin in the hippocampus were examined by immunohistochemistry. Results The mRNA expression of HIF-1α in the hippocampus of all the rats in the vehicle group after SE increased significantly with similar trend. Six h after SE, the HIF-1α mRNA expression significantly decreased with time prolongation （P〈0.05）. Major HIF-1α positive cells were located in different areas at different time after SE. In intervention experiment, the HIF-1αmRNA exp

关 键 词：发育鼠 癫痫持续状态 低氧诱导因子-1α NESTIN 

分 类 号：R742.1[医药卫生—神经病学与精神病学]