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目的研究巨噬细胞经典模式极化(M1极化)和替代模式极化(M2极化)过程中,细胞内活性氧(ROS)的变化。方法体外培养小鼠巨噬细胞RAW264.7细胞,分别用干扰素-γ(IFN-γ)和白细胞介素4(IL-4)诱导细胞向M1极化和M2极化。采用直接免疫荧光标记和流式细胞术分析CD86、CD16/32和CD206表达,流式细胞术检测细胞内ROS。结果 IFN-γ诱导RAW264.7细胞M1极化,对照组细胞CD86、CD16/32和CD32阳性细胞百分率分别为(56.4±4.1)%、(96.2±2.0)%和(19.4±4.3)%,而50μg/L IFN-γ诱导12 h后,3种分子的百分率分别为(67.4±5.2)%、(95.9±3.5)%和(9.7±2.0)%;IL-4诱导RAW264.7细胞M2极化,25μg/L IL-4诱导12 h后,3种分子的百分率分别为(39.1±4.4)%、(95.9±3.3)%和(23.8±4.6)%。对照组、IFN-γ(50μg/L)和IL-4(25μg/L)诱导组细胞ROS相对含量分别为(34.5±4.0)%、(66.2±5.8)%和(44.9±5.5)%。结论极化是巨噬细胞活化和功能转变的过程,伴随其M1和M2极化,细胞内ROS均增加。
Objective To study the changes of reactive oxygen species (ROS) in macrophages during the classical mode polarization (M1 polarization) and alternative mode polarization (M2 polarization). Methods Mouse macrophage RAW264.7 cells were cultured in vitro, and the cells were induced to M1 and M2 by interferon-γ (IFN-γ) and interleukin-4 (IL-4) respectively. Direct immunofluorescence labeling and flow cytometry analysis of CD86, CD16 / 32 and CD206 expression, flow cytometry intracellular ROS. Results The percentage of CD86, CD16 / 32 and CD32 positive cells in RAW264.7 cells induced by IFN-γ was (56.4 ± 4.1)%, (96.2 ± 2.0)% and (19.4 ± 4.3)%, respectively The percentages of the three molecules were (67.4 ± 5.2)%, (95.9 ± 3.5)% and (9.7 ± 2.0)%, respectively, after induced by 50μg / L IFN-γ for 12 h. IL- (39.1 ± 4.4)%, (95.9 ± 3.3)% and (23.8 ± 4.6)%, respectively, after 12 h induction with 25 μg / L IL-4. The relative content of ROS in the control group was 34.5 ± 4.0%, (66.2 ± 5.8)% and (44.9 ± 5.5)%, respectively, when induced by IFN-γ (50μg / L) and IL-4 (25μg / Conclusion Polarization is the process of macrophage activation and functional transformation. With the polarization of M1 and M2, intracellular ROS increases.