三磷酸腺苷诱导人白血病细胞U937凋亡的电镜观察

ELECTRON MICROSCOPIC OBSERVATION ON ATP INDUCED U937 CELLS APOPTOSIS

目的 为探讨三磷酸腺苷 (ATP)对人白血病细胞 U937的促凋亡作用。 方法 应用 ATP(0 .2 3g/L)作用于人白血病细胞 U937(human histocytic lym phom a) ,(1)用白细胞计数器每日计数细胞和计算抑制率 ;(2 )用流式细胞分析仪检测 ATP对 U937细胞周期改变和凋亡的影响 ;(3)苏木精 -伊红染色和电子显微镜检测凋亡小体的形成过程 ;(4)用苯胺黑染色检测凋亡小体的生命本性。 结果 (1) ATP对 U937细胞的增殖有明显的阻抑作用 ,加药 48h后增殖的抑制率可达 43%以上 ;(2 ) ATP处理的 U937细胞周期发生改变 ,G1 期细胞数明显增多 ,ATP作用 2 4h时 G1 期前出现亚二倍体峰——凋亡峰 ,凋亡细胞数为 3.4% ,作用 48h时凋亡细胞数增多为2 2 .7% ;(3) ATP处理的 U937细胞首先在核内染色质浓缩成半月形贴近核膜 ,逐渐向核膜外移动 ,进入胞浆内再移向质膜内外面 ,紧贴质膜外面再逐步脱离细胞体 ,进入细胞基质中 ,成为游离的凋亡小体。凋亡小体有双层核膜及内质网和线粒体与深染的染色质 ;(4)凋亡小体拒染黑色素。 结论 (1) ATP对 U937细胞的增殖有明显的阻抑作用 ;(2 ) ATP诱导 U937细胞凋亡 ;(3) ATP阻断 U937细胞于 G1 期 ,G1 期前出现凋亡峰并出现凋亡小体 (4)凋亡小体是有生命的结构 ;(5 ) ATP是 U937细胞凋亡?

Objective\ In order to study the effects of ATP on U937 cells (human histocytic lymphema). Methods Cell culture, flow cytometry, HE staining, Nigrosine staining and electron microscopy were used. Results (1)ATP could inhibit the proliferation of U937 cells with inhibition rate over 43% after 48hour ATP treatment; (2)ATP treated U937 cells numbers increased obviously in G 1 phase, and apoptosis peak appeared before G 1 phase, apoptotic cell number was 3 4% for 24hour, and was 22 7% for 48hour of ATP treated U937 cells;(3) Nuclear chromatin condensed into sperical masses bound cytoplasma membrane formed apoptotic bodies which is shed from membrane surface into intercellular medium;(4) Apoptotic bodies were nigrosine staining negtive. Conclusion ATP inhibited the proliferation and induced the apoptosis of U937 cells (human histocytic lymphoma). It blocked U937 cells in G 1 phase and formed apoptotic bodies. It was indicated that ATP would be a effective promoter of U937 cells apoptosis.