摘要
目的研究Nurr1基因过表达在6-羟基多巴胺(6-OHDA)选择性诱导多巴胺(DA)能神经元特异性损伤中的作用。方法①不同浓度6-OHDA作用不同时间,比较细胞形态;②细胞经6-OHDA作用后,经流式细胞术(FCM)检测细胞周期分布比例;③不同浓度6-OHDA作用不同时间,经AnnexinV/PI双染后,运用共聚焦显微镜观察细胞凋亡情况,获得毒素诱导凋亡的最适剂量(75μmol/L)和时间(12 h),运用FCM检测两株细胞早期凋亡比例。结果①经6-OHDA处理后,倒置相差显微镜观察,结果显示SK-N-SH/Nurr1细胞形态损伤早于SK-N-SH细胞,且损伤程度明显;②细胞周期结果显示,6-OHDA诱导SK-N-SH细胞G2/M期细胞比例下降,而SK-N-SH/Nurr1细胞S期细胞比例下降;③经AnnexinV/PI双染联合FCM检测早期凋亡比例结果显示,SK-N-SH/Nurr1细胞凋亡比例明显高于SK-N-SH细胞(P<0.05)。结论外源性Nurr1基因过表达促进了6-OHDA诱导的SK-N-SH/Nurr1细胞凋亡,Nurr1可能与SK-N-SH/Nurr1细胞对6-OHDA损伤敏感性增强有关。
Objective To compare the different damage mechanism between overexpressed extrinsic SK-N-SH/Nurr1 of Nurr1 and SK-N-SH induced by 6-OHDA and to explore the effect of overexpressed Nurr1 gene on selective cell apoptosis of dopamine （DA） ergic neurons induced by 6-OHDA. Methods ①Cells morphology were observed at different concentrations of 6-OHDA and different time. ②Cell cycle analysis was performed using flow cytometry （FCM） after treated by 6-OHDA. ③Cellular apoptosis treated by 6-OHDA at different time were observed by confocal microscope after Annexin V/PI staining to obtain the optimal dosage （75μmol/L） and time （ 12 h） to detect the ratio of early apoptosis of two strains with FCM. Results ① Under phase-contrast microscope, cell ratio of SK-N-SH/Nurr1 cells G2/M phase and SK-N-SH/Nurr1 S phase reduced after treated by 6-OHDA. ② Cell cycle results showed a decrease in S phase of cells in SK-N- SH/Nurr1 cells and G2/M phase in SK-N-SH cells. ③AnnexinV/PI double staining with FCM showed the cellular apoptosis ratio in SK-N- SH/Nurr1 cells were significantly higher than that of SK-N-SH cells （P 〈 0. 05 ）. Conclusions Overexpressed extrinsic Nurr1 could enhance apoptosis of SK-N-SH/Nurr1 cells. Nurr1 gene might be related to increased sensitivity of SK-N-SH/Nurr1 to 6-OHDA.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2006年第7期955-958,共4页
Chinese Journal of Gerontology
基金
北京市"科技新星"计划项目(2003A026)
北京市神经损伤修复再生研究重点实验室开放课题
北京市优秀人才培养专项经费资助项目
首都医学发展基金资助项目
