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NF—κB在永生化神经前体细胞凋亡中的作用
  • ISSN号:0254-1416
  • 期刊名称:《中华麻醉学杂志》
  • 时间:0
  • 分类:R971.1[医药卫生—药品;医药卫生—药学] R733.71[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:[1]华中科技大学同济医学院附属同济医院麻醉科,武汉市430030
  • 相关基金:国家自然科学基金(30300328)
中文摘要:

目的探讨NF-κB在永生化神经前体细胞凋亡中的作用。方法永生化神经前体细胞接种于6孔板,随机分为5组,每组6孔,INPC组不进行基因转染,INPC/CMV组转染表达质粒RcCMV;INPC/050组、INPC/p65组分别转染含050基因和p65基因的表达质粒RcCMV,转染后2d,加入200μg/ml G418筛选12~14d,进行阳性克隆扩大培养3—4周,检测050mRNA、p65mRNA表达、NF-κB活性和细胞凋亡情况。INPC/p50#5组转染含p65基因的表达质粒RcCMV,加入200μg/mlG418筛选12—14d,进行阳性克隆扩大培养3~4周,转染含pSO基因的表达质粒RcCMV,2d后进行上述指标的检测。结果INPC/050组和INPC/p50#5组p50mRNA表达高于其他组(P〈0.05),INPC/p65组和INPC/p50p65组p65mRNA表达、NF-κB活性及细胞凋亡率高于其他组(P〈0.05)。结论NF-κB活性升高可促讲永牛化神绎前体细胞的凋产.

英文摘要:

Objective To investigate the role of NF-κB in apoptosis of immortalized neural progenitor cells ( INPCs). Methods INPCs were cultured in 6-well plates and were randomly divided into 5 groups ( n = 6 each) : group I was not transfected with any plasmid (group INPC); group II was transfected with control plasmid (group INPC/CMV) ; group m was transfeeted with plasmid RcCMV-p50 ( group INPC/p50) ; group IV was transfected with plasmid RcCMV-p65 (group INPC/p65) and groupV was transfected with plasmid RcCMV-p50 and RcCMV-p65 (group INPC/p50p65). Group INPC/CMV (II), INPC/p50 (m) and INPC/p65 (IV) were screened by G418, and the positive clones were then cultured for 3-4 weeks. The transcription of p50 mRNA or p65 mRNA was detected by RT-PCR. The NF--κB activity was measured by luciferase reporter gene assay. The cell apoptosis was measured by mmexin V/PI staining. In group INPC/p50p65 and group INPC/p65, the cultured positive clone was transiently transfected with plasmid RcCMV-p50. Two days after transfection, the same measurement was performed in group INPC/p50p65 and the other groups. Results The expression of p50 mRNA was significantly increased in group INPC/p50 and INPC/p50p65 as compared with the other groups (P 〈 0.05). The expression of p65 mRNA, the NF-κB activity and the apoptotic rate were significantly increased in group INPC/p65 and INPC/p50p65 as compared with the other groups ( P 〈 0.05). Conclusion Enhanced NF-κB activity can increase immortalized neural progenitor cell apoptosis.

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期刊信息
  • 《中华麻醉学杂志》
  • 中国科技核心期刊
  • 主管单位:中国科协
  • 主办单位:中华医学会
  • 主编:
  • 地址:石家庄市和平西路299号
  • 邮编:050021
  • 邮箱:cja@vip.163.com
  • 电话:0311-85989621
  • 国际标准刊号:ISSN:0254-1416
  • 国内统一刊号:ISSN:13-1073/R
  • 邮发代号:18-49
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:38293