人早幼粒白血病细胞HL-60胞内pH的^(31)P核磁共振研究

Study on Intracellular pH of HL-60 Cells by ^(31)P Nuclear Magnetic Resonance

建立了无损伤性测定HL 60细胞胞内pH的31 P NMR方法。细胞内无机磷 (Pi)的化学位移对pH非常敏感 ,随pH变化而变化 ,通过测定其化学位移进而能间接确定细胞内的pH。HL 60细胞的31 P核磁共振谱由Pi、ATP等的共振峰组成。根据Pi 峰的化学位移对HL 60细胞内pH进行了测定。HL 60细胞内Pi 峰的化学位移为 5 .78± 0 .0 1 ,计算得到细胞内pH值为 6.1 6±0 .0 1。31 P核磁共振能在测定细胞胞内pH的同时 ,观测到细胞内多种含磷小分子代谢物 。

A 31P nuclear magnetic resonance (NMR) method has been established to noninvasively measure intracellular pH of human promyelocyte leukemia cell line HL-60. The chemical shift of intracellular inorganic phosphate (P i),which is very sensitive to cytosolic pH,shifts its position on the 31P-NMR spectrum as pH changed. Cytosolic pH can be derived from chemical shift of P i peak. Intracellular pH 6.16±0.01 was calculated based on the chemical shift of P i peak (5.78±0.01) in HL-60 cells. 31P-NMR, which can measure the intracellular pH as well as determine many kinds of phosphorus-containing metabolites, has been applied as a valuable method for the study of intracellular pH and metabolism of tumor cells in physiological condition by noninvasively monitoring phosphorus-containing compounds involved in cellular metabolism in intact actively tumor cells.