钠氢交换蛋白-1特异性抑制剂二甲基氨氯吡咪对HL-60-ADM细胞pHi变.docVIP

钠氢交换蛋白-1特异性抑制剂二甲基氨氯吡咪对HL-60/ADM细胞pHi变 　 作者：常城，孔佩艳，陈幸华，彭贤贵，刘林，刘红，曾东风，梁雪，王庆余 【摘要】 为了探讨钠氢交换蛋白-1（NHE-1）抑制剂二甲基氨氯吡咪（DMA）对HL-60/ADM细胞pHi变化及细胞增殖、凋亡的影响，以敏感HL-60细胞为对照，用不同浓度的DMA干预后，荧光指示剂（BCECF/AM）检测法检测细胞内pHi，微量噻唑蓝法（MTT）检测细胞生长的抑制率，流式细胞术检测细胞周期变化，TUNEL法检测原位细胞凋亡，对比HL-60/ADM细胞与HL-60细胞之间的差异。结果显示： HL-60/ADM细胞内pHi较HL-60细胞显著升高；DMA作用下，HL-60/ADM细胞的pHi降低幅度、生长抑制率、细胞凋亡率均显著高于HL-60细胞；当DMA浓度100-150 μmol/L时，HL-60/ADM细胞的G0/G1期细胞比率增加幅度、S期及G2/M期细胞比率降低幅度均高于HL-60细胞。 结论： NHE-1特异性抑制剂DMA引起HL-60/ADM的pHi降低，可抑制细胞生长并诱导细胞凋亡；且DMA对HL-60/ADM细胞的生长抑制作用显著高于HL-60细胞。 【关键词】 钠/氢交换蛋白-1 　　Effect of the NHE-1-Specific Inhibitor DMA on pHi，Proliferation and Apoptosis of HL-60/ADM Cells In Vitro 　　Abstract The aim of this study was to evaluate the effect of dimethyl amiloride (DMA)，a specific inhibitor of Na+/H+ exchanger-1 (NHE-1)，on intracellular pH value(pHi)，proliferation and apoptosis of HL-60/ADM cells in vitro. After treatment with DMA at different doses，pHi of HL-60 and HL-60/ADM cell lines were determined by using pH-sensitive fluorescence dye BECEF-AM; the rate of growth inhibition of cells was detected with MTT assay; cell cycle was detected by flow cytometric DNA analysis; cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The results showed that pHi in HL-60/ADM cells was higher than that in HL-60 cells. After treatment with DMA at different doses，pHi decreased，the rate of growth inhibition and the rate of apoptotic cells in HL-60/ADM cells were all higher than those in HL-60 cells. Meanwhile，after treatment with DMA during 100 μmol/L to 150 μmol/L，the increase amplitude of G0/G1 phase cells and the decrease amplitude of S＋G2/M cells in HL-60/ADM cells were higher than those in HL-60 cells. It is concluded that by causing intracellular acidification，the NHE-1-specific inhibitor DMA inhibits proliferation of HL-60/ADM cells and induces apoptosis of HL-60/ADM cells，and the degree of this growth inhibition of HL-60/ADM cells is higher than that of HL-