CAJ | 학술논문

目的研究转录因子NFATc及NF-κB在钙神经素介导的脑缺血再灌注损伤中的作用.方法 Westernblotting和EMSA分子生物学技术.结果与对照组相比较,CsA明显减低I/R组Fas配体和NFATc的蛋白表达;对照组、I/R组和CsA处理组I-κB-α蛋白表达无显著区别;未观察到对照组、I/R组和CsA处理组有phosph0-I-κB-α蛋白表达;与对照组相比较,CsA明显减低I/R组Fas配体启动子远端和Fas配体启动子近端NFAT结合位点的NFAT-DNA结合活性(P＜0.01).结论转录因子NFATc参与钙神经素介导的脑缺血再灌注损伤,促进CD95配体分子的转录表达;NF-κB可能未参与钙神经素介导的脑缺血再灌注损伤的作用机制.
목적연구전록인자NFATc급NF-κB재개신경소개도적뇌결혈재관주손상중적작용.방법 Westernblotting화EMSA분자생물학기술.결과여대조조상비교,CsA명현감저I/R조Fas배체화NFATc적단백표체;대조조、I/R조화CsA처리조I-κB-α단백표체무현저구별;미관찰도대조조、I/R조화CsA처리조유phosph0-I-κB-α단백표체;여대조조상비교,CsA명현감저I/R조Fas배체계동자원단화Fas배체계동자근단NFAT결합위점적NFAT-DNA결합활성(P＜0.01).결론전록인자NFATc삼여개신경소개도적뇌결혈재관주손상,촉진CD95배체분자적전록표체;NF-κB가능미삼여개신경소개도적뇌결혈재관주손상적작용궤제.
Aim To study the involvements of nuclear factor of activated T-cells (NFATc) and NFκB in calcineurin-mediated ischemic brain damage in vivo. Methods The rat transient forebrain ischemia conducted through 15 min ischemia followed by 8, 24, and 72 h reperfusion was induced using the fourvessel method. The rats were divided randomly into five groups; sham control group, ischemia/reperfusion (I/R) group, CsA treated groups (for 8, 24, and 72 h reperfusion). Western blotting was performed to detect changes of FasL, NFATc, I-κB-α, and phospho-I-κB-α protein expression, and gel shift assays for NFAT FasL-DNA binding activities. Results Western blotting showed that the expressions of both FasL and NFATc protein were significantly increased in the hippocanpus of rat subjected to transient forebrain ischemia in comparison with those of the sham control group, which were markedly reduced by CsA. The I-κB-α protein showed no changes in all groups, and phospho-I-κB-α protein was not observed in this study. Proximal and distal FasL promoter NFAT sites bind NFAT proteins from the hippocampal neurons subjected to transient forebrain ischemia, and DNA-binding activities increased significantly compared with those of the sham control group. CsA markedly inhibited these changes. Conclusion NFATc may be involved in calcineurin-mediated ischemic brain damage and transcription factor NF-κB may not be involved.