苦参碱对吡柔比星诱导HL-60细胞凋亡及核因子-κBp65活性的影响

Effects of matrine on apoptosis and nuclear factor kappaBp65 activity of HL-60 cells induced by pirarubicin

目的 研究苦参碱对吡柔比星诱导早幼粒细胞白血病细胞系HL-60细胞凋亡与核因子-κBp65（NF-κBp65）活性的影响.方法 3种不同浓度苦参碱、吡柔比星以及苦参碱和吡柔比星联合用药,分别作用于HL-60细胞,荧光双染法和流式细胞检测技术（FCM）分别检测HL-60细胞的凋亡率,FCM检测HL-60细胞NF-κBp65活化率,实验设空白对照组.结果 细胞凋亡率：空白对照组为（6.46±1.45）％;苦参碱0.1 μ.mol/L、1.0μmol/L、10.0 μmol/L组分别为（7.14±2.95）％、（12.34±2.55）％、（19.3±2.31）％,组间比较差异有统计学意义（F=257.72,P＜0.001）;吡柔比星1.0 μmol/L、10.0 μmol/L、100.0 μmol/L组分别为（31.78±4.31）％、（47.25±5.27）％、（56.49±1.59）％,组间及各组与空白对照组比较差异均有统计学意义（F=679.19,P＜0.001）;联合用药组为（70.17±5.68）％,与1.0 μmol/L吡柔比星组（31.78 ±4.31）％比较差异有统计学意义（t=38.94,P＜0.001）.NF-κBp65活化率：空白对照组为（8.44±2.20）％;苦参碱0.1 μmol/L、1.0μmol/L、10.0 μmol/L组分别为（8.34±1.52）％、（7.11±1.29）％、（4.78±0.31）％,组间比较差异均有统计学意义（F =65.35,P＜0.001）;吡柔比星1.0 μmol/L、10.0μmol/L、100.0 μ.mol/L组分别为（16.21 ±1.20）％、（23.98±3.21）％、（32.44±2.89）％,组间及各组与空白对照组比较差异均有统计学意义（F=674.11,P＜0.001）;联合用药组为（12.01±2.27）％,与1.0 μmol/L吡柔比星组（16.21±1.20）％比较差异有统计学意义（t=21.42,P＜0.001）.结论 苦参碱和吡柔比星均能诱导HL-60细胞凋亡,呈剂量依赖性;苦参碱抑制HL-60细胞NF-κBp65活性,吡柔比星诱导其活化;苦参碱能增强吡柔比星对HL-60细胞凋亡的诱导作用,降低吡柔比星对NF-κBp65的活化作用.

Objective To explore the effects of matrine on apoptosis and nuclear factor（NF）-κBp65 activity of HL-60 cells induced by pirarubicin （THP）.Methods Effects of the apoptosis：the HL-60 cells in blank control group were cultured 14 hours with RPMI 1640; groups of different concentration drugs：matrine：0.1 μmol/L,1.0 μmol/L,10.0 μmoL/L and THP：1.0 μmol/L,10.0 μmol/L,100.0 μmol/L.The apoptosis of HL-60 cells were tested by fluorescence double colour painting and flow cytometry （FCM）.The NF-κBp65 activity of HL-60 cells were determined by FCM.Results The rates of apoptosis of HL-60 cells were （7.14 ± 2.95） %,（12.34 ± 2.55） %,（19.3 ±2.31）% and （31.78 ±4.31）%,（47.25 ±5.27）%,（56.49 ±1.59）% in matrine 0.1 μmol/L,1.0 μmol/L,10.0 μmol/L and THP 1.0 μmoL/L,10.0 μmol/L,100.0 μmol/L groups,compared to blank control group （6.46 ± 1.45） %,there were significant difference （F =257.72,677.19 ; all P 〈 0.001） ; （70.17 ± 5.68） % in matrine 0.1 μ mol/L ＋THP 1.0 μ moL/L group[vs the THP 1.0 μmoL/L group （31.78 ±4.31）%,（t =38.94,P〈0.001）].The rates of NF-κBp65 activity of HL-60 cells in matrine 0.1 μmol/L,1.0 μmol/L,10.0 μmol/L and THP 1.0 μmol/L,10.0 μmol/L,100.0 μmol/L groups were（8.34 ± 1.52）%,（7.11 ± 1.29）%,（4.78 ±0.31）% and （16.21 ± 1.20） %,（23.98 ± 3.21） %,（32.44 ± 2.89） %,with the control group （8.44 ± 2.20） %,there were significant difference（F =65.35,P 〈 0.001 ; F =674.11,P 〈 0.001） ; （12.01 ± 2.27） % in matrine 0.1 μmol/L ＋ THP 1.0 μmol/L group [with （16.21 ± 1.20） % of THP 1.0 μmol/L group（t =21.42,P 〈 0.001）].Conclusions The apoptosis of HL-60 cell is induced by matrine and pirarubicin in a dose-dependent manner.NF-κBp65 activity of HL-60 cell is inhibited by matrine and increased by pirarubicin in a dose-dependent manner.Matrine can enhannce the effect of induction by pirarubicin of apoptosis of HL-60 cell,and decrease activity of NF-κBp65 by pirarubicin.