摘要
目的 探讨核因子 κB( NF-κB)在谷氨酸致 PC1 2细胞兴奋毒性中的作用。方法 用突变 IκB质粒 (特异阻断 NF-κB活性 )转染 PC1 2细胞 ,分别用谷氨酸处理转染和未转染的两组 PC1 2细胞 ,间接免疫荧光 ( FITC)标记代表 NF-κB的 P65亚基 ,碘化丙啶 ( PI)标记细胞 DNA指示核区 ,激光扫描共聚焦显微镜 ( LSCM)检测 NF-κB的状态 ,用可反映活细胞的四氮唑盐测定判断细胞生存率。结果 突变 IκB质粒转染的 PC1 2细胞 ,经谷氨酸处理后 ,易位入核的指示 NF-κB的绿色荧光明显少于未转染组 ,细胞生存率明显低于未转染组 ;而未转染组 PC1 2细胞 ,在经谷氨酸处理后核内 NF-κB明显增多。结论 谷氨酸可导致 PC1 2细胞死亡 ,NF-κB活化是 PC1 2细胞对谷氨酸兴奋毒性的保护反应 ,可减轻谷氨酸兴奋毒性损伤。
Objective To investigate the role of NF-κB in glutamate induced excitotoxicity of PC12 cell. Methods A mutant IκBα(S 32,36→A) was transfected into PC12 cells by lipofectamine tranfection and its transient effect was examined 48 hours after transfection. The transfected PC12 and untransfected C12 were cultured with glutamate respectively.The activation of NF-κB was detected by laser scanning confocal microscopy with FITC-tagged p65 antibody and PI-staining DNA. Cell survival was detected with Wst-1 reagent. Results NF-κB/p65 associated fluorescence can be seen in the nuclei of untransfected P cells after glutamate stimulation, no such fluorescence were observed in the nuclei of transfected cells. The cell survivals of untransfected PC12 cells after glutamate stimulation were significantly higher than that of transfected PC12 cells (P<0.01). Conclusions These results demonstrate that glutamate may cause PC12 cells death. The activation of NF-κB may be one of the mechenisms through which PC12 cells avoid glutamate induced excitotoxicity. NF-κB activation may play a neuroprotective role.
出处
《中国神经免疫学和神经病学杂志》
CAS
2003年第3期194-196,203,T002,共5页
Chinese Journal of Neuroimmunology and Neurology
基金
国家自然科学基金资助项目 ( 3 93 3 0 2 10 )
