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目的:以人原髓细胞白血病细胞HL-60为例,观察左旋-二脱水伊地醇双甲磺酸酯(DMDAI-L)对HL-60细胞的诱导凋亡作用,初步探讨其作用机制。方法:通过酶解可见光底物DEVD-p NA测定给药前后HL-60细胞内的caspase-3酶活性;膜电位依赖性结合的荧光探针JC-1标记法观察给药前后细胞线粒体膜电位(△Ψm)的荧光变化。结果:DMDAI-L 1.25,2.5,5,10和20μg·ml-1剂量组作用24 h后,HL-60细胞内的caspase-3酶活性显著上升(P<0.05);在DMDAI-L作用下,随着浓度增加及作用时间延长,HL-60细胞内线粒体膜电位明显降低。结论:DMDAI-L诱导HL-60细胞凋亡过程中,其作用机制可能与在一定剂量范围及作用时间内,激活或调节细胞内的caspase-3酶活性、降低细胞线粒体膜电位有关。
OBJECTIVE: To investigate the apoptosis-inducing effect of DMDAI-L on HL-60 cells using human promyelocytic leukemia HL-60 as an example, and to explore its mechanism. Methods: The activity of caspase-3 in HL-60 cells before and after administration was determined by enzymatic digestion of visible light substrate DEVD-p NA. The membrane potential-dependent binding of JC-1 fluorescent probe was used to observe the mitochondrial membrane potential (△ Ψm) fluorescence changes. Results: The caspase-3 activity in HL-60 cells increased significantly (P <0.05) after DMDAI-L 1.25,2.5,5,10 and 20μg · ml-1 for 24 h. Under the action of DMDAI-L The mitochondrial membrane potential in HL-60 cells was significantly decreased with the increase of concentration and time. CONCLUSION: The mechanism of DMDAI-L inducing HL-60 cell apoptosis may be related to activating or regulating the activity of caspase-3 and decreasing the mitochondrial membrane potential in a dose and time-dependent manner.