首都医科大学学报 ›› 2012, Vol. 33 ›› Issue (2): 182-186.doi: 10.3969/j.issn.1006-7795.2012.02.010

• 基础研究 • 上一篇    下一篇

抑制低氧诱导因子-1α对PC12细胞存活的影响

李冉1, 王勇1, 宫晓丽1, 范明2, 王晓民1, 朱玲玲2, 汪璇1   

  1. 1. 首都医科大学基础医学院生理学系,教育部神经变性病重点实验室,北京 100069;2. 军事医学科学院基础医学研究所认知科学研究室,北京 100850
  • 收稿日期:2012-01-04 修回日期:1900-01-01 出版日期:2012-04-21 发布日期:2012-04-21
  • 通讯作者: 汪 璇

Effect of echinomycin on the cell viability of PC12 cells after it inhibits hypoxia inducible factor-1α

LI Ran1, WANG Yong1, GONG Xiao-li1, FAN Ming2, WANG Xiao-min1, ZHU Ling-ling2, WANG Xuan1   

  1. 1. Department of Physiology, School of Basic Medical Science, Capital Medical University, Key Laboratory for Neurodegenerative Disorders of the Ministry of Education, Beijing 100069, China;2. Department of Cognitive Sciences, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China
  • Received:2012-01-04 Revised:1900-01-01 Online:2012-04-21 Published:2012-04-21

摘要: 目的 通过抑制低氧诱导因子-1α转录活性,研究低氧诱导因子-1α在低氧诱导的细胞死亡中的作用。方法 1 将PC12细胞接种于12孔板,在20%常氧和3%低氧环境中培养24 h,在光镜下拍照并应用计数方法记录PC12细胞的生长情况。2 将PC12细胞接种于96孔板,在常氧和低氧环境中培养24 h后,用细胞活力检测剂(a novel tetrazolium compound, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt,MTS)检测PC12细胞的生长情况。3 在常氧和低氧条件下分别用5 nmol/L、50 nmol/L、500 nmol/L的低氧诱导因子-1α抑制剂棘霉素处理PC12细胞24 h和48 h后,用MTS法检测细胞活力。结果 1 细胞计数和MTS法检测细胞活力的结果均显示3%的低氧条件可明显诱导PC12细胞死亡。2 不同浓度的低氧诱导因子-1α抑制剂棘霉素处理PC12细胞24 h之后,低氧和常氧的细胞活力没有显著区别。3 不同浓度的低氧诱导因子-1α抑制剂棘霉素处理PC12细胞48 h之后,低氧不再促进细胞死亡,相反,低氧条件处理的细胞活力明显好于常氧。结论 低氧会促使PC12细胞死亡,棘霉素抑制HIF-1α的转录活性24 h后,低氧对PC12细胞的促死亡作用减弱;48 h后,低氧组的细胞活力还可超过常氧组。这表明,低氧下过量的HIF-1α转录激活对细胞是有害的。

关键词: 低氧, 低氧诱导因子1α, PC12细胞, 棘霉素

Abstract: Objective To clarify the role of hypoxia inducible factor(HIF)-1α in the process of hypoxia-induced cell death through inhibiting its transcription activity. Methods 1 PC12 cells were seeded in the 12-well plate and treated for 24 h in normoxia(20% O2) or hypoxia(3% O2). Then photomicrographs under light microscope were taken and analyzed for situation of cell growth by cell-counting method. 2 PC12 cells were seeded in 96-well plate and treated for 24 h in 20% O2 or 3% O2. A test for cell survival called MTS was further used to detect the cell viability of PC12 cells. 3 PC12 cells were treated with HIF-1α inhibitor-echinomycin for 24 h or 48 h at 5 nmol/L, 50 nmol/L and 500 nmol/L, respectively. Then MTS was used to detect the cell viability. Results 1 3% O2 induced the death of PC12 cells significantly contrary to 20% O2. 2 Echinomycin administration eliminated the cell survival difference between 20% O2 and 3% O2 at 24 h. 3 After a 48 h-treatment with echinomycin, it even reversed the hypoxia-enhanced cell death relative to normoxia. Conclusion Hypoxia could promote the death of PC 12 cells. After we used HIF-1α inhibitor echinomycin for 24 h, the death-promotion role of hypoxia was weakened. It even reversed the hypoxia-enhanced cell death after a 48 h-treatment with echinomycin. All these data suggested that the excessive transcription activation during hypoxia was detrimental to PC 12 cells.

Key words: hypoxia, hypoxia inducible factor-1α, PC12 cells, echinomycin

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