超氧阴离子参与白藜芦醇诱导的人急性早幼粒白血病HL-60细胞凋亡

Superoxide Anions Involved in the Apoptosis of Human Promyelocytic Leukemia HL-60 Cells Induced by Resveratrol

目的:研究超氧阴离子(O2ˉ.)、H2O2、线粒体膜电位(MMP)在白藜芦醇(RSV)诱导人急性早幼粒白血病HL-60细胞凋亡中的作用。方法:以RSV(10.0、50.0、100.0μmol/L)分别作用HL-60 6、12、24h后,用流式细胞术检测细胞内O2ˉ.、H2O2及MMP水平;四氮唑蓝凝胶法检测RSV作用HL-6024h后超氧化物歧化酶(SOD)的活性;MTT法检测SOD和RSV共同作用HL-60后的细胞生存率;锥虫蓝细胞计数法检测N-乙酰-L-半胱氨酸(NAC)和RSV共同作用HL-60后的细胞生存率。结果:RSV可使HL-60内O2ˉ.水平升高,H2O2水平、MMP降低,SOD活性无明显变化;SOD和NAC对RSV诱导的HL-60凋亡作用无明显影响。结论:RSV诱导HL-60凋亡机制是RSV引起细胞内O2ˉ.水平升高,MMP降低,从而启动caspases凋亡途径诱发细胞凋亡。

Objective： To investigate the effects of superoxide anion（O2^-）, hydrogen peroxide（H2O2）, mitochondrial membrane potential （MMP）on the apoptosis of human promyelocytic leukemia HI-60 ceils induced by resveratrol （RSV）. Methods： HL-60 coils were treated with RSV（10.0, 50.0, 100.0 μmol/L）for 6, 12 and 24 hours. The flow cyometry was performed to test the level of O2^-, H202 and MMP. The NBT-gel method was used to detect the activity of superoxide dismutase（ SOD） of HL-60 colls after treatment with RSV for 24 hours. The MTT assay was used to detect the cell viability of HL-60 cells after treatment with the combination of RSV and SOD. The trypan blue staining was performed to detect the cell viability of HL-60 cells after treatment with the combination of RSV and N-Acetyl-L-cysteine （NAG）. Results： The production of O2^-· of HL-60 cells was raised after treatment with RSV, but the levels of H202 and MMP were reduced and the activity of SOD wasn＇t changed significantly. SOD and NAC had no effects on the apeptosis of HL-60 ceils induced by RSV. Conclusion： The mechanism of the apeptosis of HL-60 cells induced by RSV is upregulation of O2^-· whilst down-regulation of MMP followed by caspase-mediated apoptotic pathway.