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研究双苯氟嗪对Fas配体分子表达抑制的基因转录机制的影响。通过四血管阻断法建立大鼠全脑缺血再灌注损伤模型,所有实验动物缺血15 min,再灌注72 h。实验大鼠随机分为4组:假手术组、缺血再灌注组、双苯氟嗪组及环孢素A组。药物干预于再灌注后2 h内给药,每日1次,连续3 d。双苯氟嗪按20 mg·kg-1灌胃给药,环孢素A 10 mg·kg-1腹腔注射。应用蛋白质印迹和电泳迁移率改变分析技术检测海马CAI区Fas配体(FasL)、转录因子NFATc、I-κB-α、phospho-I-κB-α蛋白表达以及测定FasL分子启动子远端及FasL分子启动子近端NFATFasL-DNA结合活性。结果表明,双苯氟嗪明显降低FasL、NFATc的蛋白表达并显著减少FasL分子启动子远端及FasL分子启动子近端NFAT结合位点的NFAT-DNA结合活性。各组之间I-κB-α蛋白表达无显著区别。未观察到各组phospho-I-κB-α蛋白表达。由此可见,双苯氟嗪通过降低转录因子NFATc的FasL分子的基因转录功能,从而抑制FasL分子的基因表达。
To study the effect of dipfluzine on the gene transcription mechanism of Fas ligand molecule expression inhibition. The model of global cerebral ischemia-reperfusion injury was established by four-vessel occlusion. All the experimental animals were ischemia for 15 min and reperfused for 72 h. The rats were randomly divided into 4 groups: sham operation group, ischemia reperfusion group, dipfluzine group and cyclosporine A group. Drug intervention was given within 2 h after reperfusion, once daily for 3 days. Dipfluzine 20 mg · kg-1 intragastric administration, cyclosporin A 10 mg · kg-1 intraperitoneal injection. Western blotting and electrophoretic mobility shift assay were used to detect FasL, NFATc, I-κB-α and phospho-I-κB-αprotein expression in hippocampal CAI region, and to detect FasL promoter and FasL Molecular Promoter Proximal NFATFasL-DNA Binding Activity. The results showed that dipfluzine significantly decreased the expression of FasL and NFATc and significantly decreased the NFAT-DNA binding activity of the FasL promoter and the proximal NFAT binding site of FasL promoter. There was no significant difference in the expression of I-κB-α between the groups. No phospho-I-κB-α protein expression was observed in all groups. Thus, dipfluzine inhibits the gene expression of FasL molecules by reducing the gene transcriptional activity of the FasL molecule of the transcription factor NFATc.