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目的探讨荧光探针JC-1在检测细胞凋亡早期线粒体膜电位(△Ψm)中的应用。方法2-甲氧基雌二醇(2-ME)诱导骨髓增生异常综合征细胞株 MUTZ-1细胞凋亡,利用新型荧光探针 JC-1标记 MUTZ-1细胞,在 FACS Calibur 流式细胞仪(FCM)上检测细胞内 JC-1荧光强度的变化;在荧光显微镜下观察和计数细胞内 JC-1荧光颜色的改变。结果对照组 MUTZ-1细胞线粒体△Ψm维持较高,JC-1在线粒体内形成聚合物,发出橘红色荧光;用药组 MUTZ-1细胞线粒体△Ψm下降,JC-1聚合物分解成单体,发出绿色荧光。MUTZ-I 细胞线粒体△Ψm的下降随着时间延长而逐渐增强,与对照组相比差异有统计学意义(P<0.01);随着药物浓度的增加,细胞线粒体△Ψm下降增强,在4 μmol/L时线粒体△Ψm下降达到最大值,以后线粒体ΔΨm下降趋势减缓。结论荧光探针 JC-1结合 FCM和荧光显微镜可以准确、直观地检测凋亡早期线粒体ΔΨm的变化,为2-ME 诱导 MUTZ-1细胞凋亡机制的研究提供了重要手段。
Objective To investigate the application of fluorescent probe JC-1 in the detection of early mitochondrial membrane potential (△ Ψm). Methods The apoptosis of myelodysplastic syndrome cell line MUTZ-1 was induced by 2-methoxyestradiol (2-ME). MUTZ-1 cells were labeled with a new fluorescent probe JC-1 and expressed in FACS Calibur flow cytometry (FCM) was used to detect the change of intracellular JC-1 fluorescence intensity. The change of intracellular JC-1 fluorescence color was observed under a fluorescence microscope. RESULTS: The mitochondrial △ Ψm of MUTZ-1 cells maintained high in the control group. JC-1 formed a polymer in the mitochondria and emitted orange-red fluorescence. The mitochondrial △ Ψm of MUTZ-1 cells decreased and the JC-1 polymer decomposed into monomers, Glowing green fluorescence. The decrease of mitochondrial △ Ψm in MUTZ-1 cells gradually increased with time, and the difference was statistically significant compared with the control group (P <0.01). With the increase of drug concentration, the mitochondrial △ ψm decreased, / L, the mitochondrial △ Ψm decreased to a maximum value, mitochondrial ΔΨm decreased after the decline. Conclusion The fluorescent probe JC-1 combined with FCM and fluorescence microscope can accurately and directly detect the change of mitochondrial ΔΨm during the early apoptosis, providing an important means for the study of the mechanism of 2-ME in inducing the apoptosis of MUTZ-1 cells.