摘要
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 explore the function of fluorescent probe JC-1 in detecting the changes of mitochondrial membrane potential （△ψm） in early apoptotic cells. Methods After 2-ME was used to induce MUTZ-1 cell apoptosis, cells were dyed with fluorescent probe JC-1, and then the changes of △ψm in the early stage of apoptotic cells were analyzed by flow cytometry or detected under fluorescent microscope. Results The control cells with high △ψm are those forming JC-1 aggregates in the inner membrane of mitochondria,thus showing orange-red fluorescence. 2-ME caused decrease of △ψm in MUTZ-1 cells, in which JC-1 maintains monomeric form,thus showing only green fluorescence. The decreases of △ψm were in a time-dependent manner,which were significantly higher than those in control group （P 〈 0.01 ）. With the increase of 2-ME, △ψm was decreased and reached plateau levels at concentration of 4 μmol/L. Conclusions Fluorescent probe JC-1 associated with FCM and fluorescent microscope is an accurate and direct method to detect the changes of △ψm in early apoptotic cells, and is an effective method for evaluating the mechanism of 2-ME-induced apoptosis in MDS （ MUTZ-1 ） cells.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2007年第8期923-926,共4页
Chinese Journal of Laboratory Medicine
基金
国家自然科学基金(3007018)
