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目的探讨过氧化氢(H2O2)对三氧化二砷(As2O3)诱导人类伯基特氏淋巴瘤细胞凋亡的影响及双染法荧光显微技术在检测细胞凋亡中的作用。方法应用As2O3诱导人类伯基特氏淋巴瘤细胞株BJAB细胞凋亡,采用Hoechst 33342和碘化丙啶(PI)核染料双染法荧光显微镜和透射电镜对细胞死亡进行观察及定量分析。结果BJAB细胞经As2O3处理后,荧光显微镜下可见大多数细胞出现细胞染色质凝集、核边集、核碎裂等凋亡的特征性改变;在低剂量(200 μmol/L)H2O2的影响下,抑制了As2O3诱导的细胞凋亡,降低细胞的死亡率;细胞的死亡方式由凋亡转变为核固缩性坏死,表现为既无凋亡的特征性改变,又非典型的细胞坏死,死亡细胞的细胞核呈固缩状态而非肿胀。结论低浓度(200 μmol/L)H2O2可抑制临床治疗浓度的As2O3诱导的人类伯基特氏淋巴瘤BJAB细胞凋亡;双染法荧光显微镜技术不但可明确区分细胞凋亡及坏死,还可判断早期凋亡细胞和晚期凋亡细胞,并可进行定量分析,是一种良好的细胞凋亡检测方法。
Objective To investigate the effect of hydrogen peroxide (H2O2) on the apoptosis of human Burkitt’s lymphoma cells induced by As2O3 and the effect of double staining fluorescence microscopy in the detection of apoptosis. Methods Apoptosis of human Burkitt ’s lymphoma cell line BJAB was induced by As2O3. Cell death was observed and quantitatively analyzed by Hoechst 33342 and propidium iodide (PI) double staining with fluorescence microscopy and transmission electron microscopy. Results The apoptosis of BJAB cells treated with As2O3 was observed under a fluorescence microscope. Most of the cells showed the characteristic changes of apoptosis such as chromatin condensation, nucleus margins and nuclear fragmentation. Under the influence of low dose of 200 μmol / L H2O2, Inhibit As2O3-induced apoptosis and reduce cell death; cell death from apoptosis to nuclear shrinkage necrosis, showed no characteristic changes in apoptosis, but also typical of cell necrosis, dead cells The nucleus is in a deflated state rather than swollen. Conclusions H2O2 at low concentration (200 μmol / L) can inhibit the apoptosis of BJAB cells induced by As2O3 in human Burkitt’s lymphoma treated with As2O3. The dual-staining fluorescence microscopy can not only distinguish between apoptosis and necrosis, but also determine Early apoptotic cells and late apoptotic cells, and quantitative analysis, is a good method of detection of apoptosis.