摘要
目的探讨2-甲氧基雌二醇（2ME2）对全脑缺血大鼠缺氧诱导因子-1α（HIF-1α）及凋亡相关基因的作用。方法将成年雄性SD大鼠168只按随机数字表法分为全脑缺血组（n=84）和全脑缺血2ME2干预组（n=841．然后按再灌注时间不同又分为6h、12h、24h、48h、72h、5d和7d7个亚组。采用改良的Pulsineli4-VO法制作全脑缺血大鼠模型。应用Nissl染色进行海马神经元计数，免疫组化及RT。PCR技术进行HIF-1α、caspase-3蛋白及RTP801mRNA表达检测。结果全脑缺血2ME2干预组48h至7d亚组神经元计数分别为37．09±3．52、26．93±3．10、22．22±3．091、6．98±3．07．与全脑缺血组相应时间点亚组相比明显增加，差异均有统计学意义（P〈0．05）。全脑缺血2ME2干预组48h至7d亚组HIF-1α、caspase-3蛋白表达阳性细胞计数分别为11．47±1．98、20．27±2．07、3．12±0．89、1．07±0．83和12．39±1．67、20．65±2．01、15．61±1．26、6．57±1．12，与全脑缺血组相应时阃点亚组相比明显减少，差异均有统计学意义（P〈0．05）。全脑缺血2ME2干预组12h至5d亚组RTP801mRNA表达吸光度比值分别为0．750±0．078、1．008±0．090、0．717±0．072、0．431±0．047、0．231±0．028，与全脑缺血组相应时间点亚组相比明显减低，差异均有统计学意义（P〈0．05）。结论在全脑缺血急性期，2ME2抑制HIF-1α及凋亡相关基因RTP801、caspase-3的表达，具有-定的神经保护作用。
Objective To investigate the effect of 2-methoxyestradiol （2ME2） on the expressions of hypoxia-inducible factor 1α and apoptosis-related genes （RTP801 and caspase-3） in the hippocampus of rats following global cerebral ischemia. Methods A total of 168 adult male SD rats were randomized into 2 groups, global cerebral ischemia group （GI group, n=84） and global cerebral ischemia＋2ME2 treatment group （GI＋2ME2 group, n=84）. In GI and GI＋2ME2 groups, 4 vessel occlusion （4-VO） global ischemia was induced, and the rats were sacrificed at 6, 12, 24, 48, 96 h, and 5 and 7 days after the reperfusion. Nissl staining was used for quantitative analysis of the hippocampal neurons, and immunohistochemistry and RT-PCR were performed to detect the expressions of HIF-1α protein, caspase-3 protein and RTP801 mRNA. Results At 48, 96 h and 5 and 7 days after the reperfusion, the numbers ofhippocampal neurons in GI＋2ME2 group were 37.09±3.52, 26.93±3.10, 22.22±3.091, and 6.98±3.07, respectively, significantly higher than those in GI group （P〈0.05）. 2ME2 significantly suppressed the expression levels of HIF-1α and reduced the numbers of the cells positive for HIF-1α protein to 11.47±1.9, 20.27±2.07, 3.12±0.89, 1.07±0.83 at these time points （P〈0.05）. The expressions of caspase-3 protein were decreased significantly in GI＋2ME2 group, with the numbers of positive cells of 12.39±1.67, 20.65±2.01, 15.61±1.26, and 6.57±1.12 at the time points. The absorbance of RTP801 mRNA expression at the time points from 12 h to 5 days in GI＋2ME2 group was 0.750± 0.078, 1.008±0.090, 0.717±0.072, 0.431 ±0.047, and 0.231 ±0.028, respectively, significantly lower than that in GI group （P〈0.05）. Conclusion 2ME2 offers brain protection in rats with global cerebral ischemia and suppresses the elevation in the expressions of hypoxia-inducible factor in, RTP801 and caspase-3.
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2009年第9期884-888,共5页
Chinese Journal of Neuromedicine
基金
基金项目：山东大学科委汁划专项（0510821
