补骨脂素加长波紫外线照射对白血病K562细胞杀伤作用.doc

补骨脂素加长波紫外线照射对白血病K562细胞杀伤作用 　 作者：张德杰，黄世林，陈楠楠，向阳，杨佩满，赵瑾瑶 【关键词】 补骨脂素 ［摘要］ 目的：探讨补骨脂的提纯物补骨脂素（psoralen）加长波紫外线（ultravioletA light）照射光化学疗法，亦称PUVA（psoralen plus ultravioletA light）疗法，对慢性粒细胞白血病红白病变细胞株K562细胞的杀伤作用。方法：以传代培养的K562细胞为模型，采用四甲基偶氮唑盐比色法（monotetrazolium test, MTT）测定单纯补骨脂素、单纯紫外线照射及PUVA疗法对白血病细胞的杀伤作用；采用流式细胞仪对其进行DNA含量分析，并用早期凋亡试剂盒AnnexinⅤFITC和碘化丙啶（propidine iodide, PI）双染法测定单纯补骨脂素对白血病细胞的作用；采用电镜技术对其进行形态学观察。结果：单纯补骨脂素或单纯紫外线照射对K562细胞均有杀伤作用，PUVA比单纯补骨脂素或单纯紫外线照射对K562细胞的杀伤作用有明显增强（P＜0.05)。PUVA处理后的白血病细胞通过流式细胞仪可以检测到亚二倍体凋亡峰。电镜下可见：细胞体积变小，胞浆浓缩，胞核染色质聚集或边集，核小体碎裂，大部分细胞呈典型的凋亡形态。结论：中药补骨脂提纯物补骨脂素对人白血病细胞K562具有杀伤作用。补骨脂素具有光敏活性，结合360 nm波长紫外线照射治疗对K562细胞的杀伤作用有明显增强。紫外线照射时间以10 min为最佳；补骨脂素适宜的终浓度为40 μg/ml。PUVA对人白血病细胞K562杀伤作用的机制主要是诱导细胞凋亡。 　　［关键词］ 补骨脂素; 光化学疗法; 白血病, 髓样, 慢性; K562细胞系; 细胞凋亡 　　ABSTRACT Objective: To observe the effects of psoralen plus ultravioletA light (PUVA) on K562 cells and the relative mechanism. Methods: The effects of psoralen, ultravioletA light and PUVA on K562 cells were assayed by monotetrazolium test (MTT). DNA content was analyzed by flow cytometry (FCM). The apoptotic rates of K562 cells treated with 40 and 80 μg/ml psoralen for 24 and 48 hours were assayed by AnnexinVFITC/PI reagent kit on FCM respectively. The ultrastructures of apoptotic cells were observed by a transmission electron microscope (TEM). Results: Either single psoralen therapy or single ultravioletA irradiation had inhibiting effect on K562 cells. The inhibiting effect of PUVA on K562 cells was stronger than that of the single psoralen therapy or single ultravioletA light irradiation (P＜0.05). Apoptotic peak (AP) was detected by FCM. TEM test showed that K562 cells treated with PUVA were smaller, having condensed cell nucleus, assembled chromatin, disintegrated nucleus body and the majority of the cells appeared to be apoptotic conformation. Conclusion: Psoralen has inhibiting effect on K562 cells, and the effect of PUVA is more significant. It is suggested that 10 min irradiation and 40 μg/ml terminal concentrati