化学学报 ›› 2006, Vol. 64 ›› Issue (11): 1179-1182. 上一篇    下一篇

研究论文

人宫颈癌细胞(Hela)内的31P核磁共振研究

梁晓东,黄荣清*,骆传环,肖炳坤,赵焱   

  1. (中国人民解放军军事医学科学院放射与辐射医学研究所 北京 100085)
  • 投稿日期:2005-11-16 修回日期:2006-02-27 发布日期:2006-06-15
  • 通讯作者: 黄荣清

Study on Intracellular of Hela Cells by 31P NuclearMagnetic Resonance

LIANG Xiao-Dong, HUANG Rong-Qing*, LUO Chuan-Huan, XIAO Bing-Kun, ZHAO Yan   

  1. (Institute of Radiation Medicine, Academy of Military Medical Sciences,
    The Peoples Liberation Army of China, Beijing 100850)
  • Received:2005-11-16 Revised:2006-02-27 Published:2006-06-15
  • Contact: HUANG Rong-Qing

建立了无损伤性31P NMR研究细胞内物质的实验方法, 并对人宫颈癌细胞(Hela)的31P NMR谱中含磷小分子代谢物的谱峰进行了分析; 细胞内无机磷(Pi)的化学位移对pH非常敏感, 通过测定其化学位移可间接确定细胞内的pH, Hela细胞内Pi峰的化学位移为5.88±0.01 (n=3), 计算得到细胞内 pH值为7.05±0.01; 通过测量Hela细胞的31P NMR谱中ATP的α磷和β磷及γ磷的化学位移差值, 得出Hela细胞内Mg2+与ATP结合的复合物MgATP和整个ATP量的比值, 计算得到Hela细胞内游离Mg2+浓度为(253.3±0.13) mmol/L (n=3), 与其它分析方法相比, 31P NMR测定细胞内游离Mg2+浓度具有对细胞样品无损伤的优点.

关键词: 31P NMR, 人宫颈癌细胞, 化学位移, 游离镁离子

A 31P nuclear magnetic resonance (NMR) method has been established to noninvasively measure intracellular condition of Hela cell, stable 31P NMR spectra of living cells were obtained and intracellular metabolite in Hela of phosphorized small molecule cell was analyzed. The chemical shift of intracellular inorganic phosphate (Pi) ,which is very sensitive to cytosolic pH, can be shifted on the 31P NMR spectrum as pH is changed and cytosolic pH can be derived from chemical shift of Pi peak. Intracellular pH 7.05±0.01 was calculated based on the chemical shift of Pi peak (5.88±0.01) in Hela cells.Based upon a measurement of chemical shift differences between the α and β-phosphoryl resonances in the 31P NMR spectrum of intracellular adenosine triphosphate (ATP) in Hela cell, the fraction of total ATP complexed to Mg2+was estimated, while cytosolic free magnesium ion concentration in Hela cells was measured nondestructively to be 253.3 mmol/L. 31P nuclear magnetic resonance method has noninvasive virtue in measuring intracellular Mg2+ compared with other technique.

Key words: 31P NMR, Hela cell, chemical shift, free magnesium