N-乙酰半胱氨酸对氢醌抑制HL-60细胞分化的保护作用

Effect of N-acetylcysteine on the suppression of HL-60 cells differentiation induced by hydroquinone

目的探讨N-乙酰半胱氨酸(NAC)在氢醌(HQ)抑制HL-60细胞向单核系分化过程中所起的作用。方法将NAC 1、2.5、5和10 mmol/分别与HQ 50μmol/L和豆蔻酰佛波乙酯(TPA)20 nmol/L共同处理细胞,处理48 h后收集细胞。通过硝基四氮唑蓝还原反应鉴定细胞分化;CCK-8检测细胞增殖;活性氧检测试剂盒鉴定活性氧的变化。结果与HQ和TPA+HQ组比较,随NAC剂量的增加可以引起细胞增殖的增加,差异具有统计学意义(P<0.01)。在HL-60细胞向单核系分化的过程中,与TPA+HQ组比较,细胞的分化能力随着NAC剂量的增加而增强,差异具有统计学意义(P<0.01),其中NAC 5和10 mmol/L作用最为显著。细胞内活性氧(ROS)检测显示,与空白对照组比较,HQ组和TPA+HQ组含量明显增强,差异有统计学意义(P<0.01);与TPA+HQ组比较,NAC各个剂量组ROS含量明显降低(P<0.01)。5个时间点测量细胞内活性氧含量差异有统计学意义(F=564.3,P<0.01)。结论 HQ能够抑制HL-60向单核系分化,这种抑制作用会随着NAC剂量的增加而降低,这种影响是通过清除细胞内活性氧来实现的。

Objective To study the role of N-acetylcysteine（NAC） in the process of inhibition of monocytic differentiation in HL-60 cells induced by hydroquinone（HQ） differentiated to monocytes.Methods HL-60 cells were co-treated with 1-10 mmol /L of NAC,50 μmol /L of HQ and 20 nmol /L of cardamom acid phorbol ester（TPA）,and then cells were collected after cultured for 48 h.Differentiation was assessed by the reduction of nitro blue tetrazolium assay;cell proliferation was detected by CCK-8;the change of reactive oxygen species was assessed by reactive oxygen species test kit.Results Compared with the HQ group and TPA ＋ HQ group,cell viability was enhanced with the addition of NAC,the difference was statistically significant（P0.01）.In the process that HL-60 cells differentiated to monocytes,compared with the TPA＋HQ group,cell differentiation was enhanced with the addition of NAC,the difference was statistically significant（P0.01）,among which NAC 5 mmol/L and NAC 10 mmol/L had the most significant difference.ROS detection showed that,compared with the control group,the intracellular reactive oxygen species of the HQ group and the TPA＋HQ group were significantly enhanced（P0.01）;compared with the TPA＋HQ group,the intracellular reactive oxygen species of NAC groups at all doses were decreased significantly（P0.01）.At 5 different time points,the intracellular reactive oxygen species levels were significantly different,F=564.3,P0.01.Conclusion HQ can inhibit HL-60 differentiation to monocytes.This inhibition would decrease with the increasing of the NAC＇s dose,and this effect comes to achieve by clearing the intracellular reactive oxygen species.