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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 To investigate the pro-apoptotic effects of adenosine triphosphate (ATP) on human leukemia cell line U937. Methods ATP (0.23g/L) was used to treat human leukemia cell line U937 (human histocytic lym phom a). (1) The cells were counted daily with an anticytometer and the inhibition rate was calculated. (2) Detection by flow cytometry Effects of ATP on cell cycle change and apoptosis of U937 cells; (3) Hematoxylin-eosin staining and electron microscopy for the formation of apoptotic bodies; (4) Aniline black staining for the detection of apoptotic bodies. Results (1) ATP inhibited the proliferation of U937 cells significantly. The inhibition rate of proliferation after 48 hours of dosing was more than 43%. (2) The cell cycle of U937 changed after ATP treatment and the number of cells in G1 phase increased significantly. The hypodiploid peak-apoptotic peak appeared at the G1 phase and the number of apoptotic cells was 3.4% at 24 hours after ATP treatment. The number of apoptotic cells increased by 22.7% at 48 hours; (3) ATP-treated U937 The cells first concentrate in the nuclear chromatin into a half-moon shape close to the nuclear membrane, gradually move out of the nuclear membrane, and then enter the cytoplasm and then move to the outside of the plasma membrane, close to the plasma membrane and then gradually leave the cell body, into the cell matrix, Become a free apoptotic body. The apoptotic body has a double nuclear membrane and endoplasmic reticulum, and mitochondria and hyperchromasia; (4) apoptotic bodies do not stain melanin. Conclusions (1) ATP has a significant inhibitory effect on the proliferation of U937 cells; (2) ATP induces apoptosis in U937 cells; (3) ATP blocks U937 cells in G1 phase, and G1 phase shows apoptosis and apoptosis. The body (4) apoptotic body is a living structure; (5) ATP is the apoptosis of U937 cells?