姜黄素对LPS活化小鼠巨噬细胞活性氧影响

Effect of curcumin on ROS in LPS-activited mice macrophage

目的研究低浓度姜黄素(curcumin)对脂多糖(lipopolysaccharide,LPS)活化巨噬细胞产生活性氧(reactive oxygen species,ROS)和发生凋亡的影响。方法体外培养小鼠巨噬细胞RAW264.7,用LPS(0、0.5、2和8μg/m L)诱导细胞,或采用低浓度姜黄素(7.5μmol/L)与LPS同时作用于巨噬细胞;流式细胞术分析活性氧(ROS)、线粒体膜电位(mitochondrial membrane potential,MMP)和细胞凋亡。结果随LPS浓度增加,小鼠巨噬细胞内ROS增加;高浓度的LPS导致细胞线粒体功能损伤和凋亡;与对照组比较,姜黄素处理后,细胞内活性氧由(8.1±0.8)下降到(6.6±1.3),差异有统计学意义(P<0.05);与LPS单独作用比较,姜黄素处理后细胞内活性氧由(61.9±5.3)下降到(47.9±3.8),差异有统计学意义(P<0.05);低浓度的姜黄素对RAW264.7细胞MMP无明显影响,但与LPS单独作用比较,姜黄素处理后细胞内MMP由(37.9±4.7)下降到(33.1±3.4),差异有统计学意义(P<0.05);低浓度姜黄素单独作用未导致发生明显凋亡或坏死,与LPS作用比较,姜黄素明显减少LPS活化后凋亡和坏死细胞的百分率,差异有统计学意义(P<0.05)。结论低浓度姜黄素降低了ROS而减轻LPS活化小鼠巨噬细胞的凋亡。

Objective To study the effect of low concentration curcumin on reactive oxygen species （ROS） produc- tion and apoptosis in lipopolysaccharide （ LPS）-activated mice macrophages. Methods Mice macrophage RAW264. 7 cells were cultured in vitro and administered to LPS at different dosages（0,0. 5,2,and 8 μg/mL）with or without low dose curcumin（7.5 μmol/L）. ROS, mitochondrial membrane potential（ MMP）, and apoptosis were measured with flow cytometry. Results ROS in RAW264. 7 cells increased with the concentration of LPS. Higher concentration of LPS in- duced cells apoptosis and impeded the function of mitochondria. Compared with the control group, after curcumin treat- ment, intracellular ROS significantly decreased from 8.1 ± 0. 8 to 6. 6 ± 1.3 （ P 〈 0. 05 ）. Compared with LPS treatment alone,the intracellular ROS significantly decreased from 61.9 _± 5. 3 to 47. 9 ±3.8 after curcumin and LPS treatment （ P 〈 0. 05 ）. Low concentrations of curcumin had no significant effect on MMP in RAW264. 7 cells, but intracellular MMP decreased significantly from 37.9 ± 4. 7 to 33.1 ± 3.4 after curcumin and LPS treatment compared with LPS treat- ment alone（ P 〈 0. 05 ）. Low dose curcumin treatment alone did not cause significant apoptosis or necrosis of RAW264. 7 cells without LPS activation, but significantly reduced the percentage of apoptosis or necrosis of the cells with LPS activa- tion （ P 〈 0. 05 ）. Conclusion Low dose curcumin reduces ROS production in LPS-activated mice macrophages.