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目的比较体外诱导巨噬细胞胞外诱捕网(MET)形成的不同方法。方法采用(0.5、1、5、10)μg/m L脂多糖(LPS)、(10、25、50、80)μmol/L佛波酯(PMA)和(50、100、150)μg/m L二氧化硅(Si O2)处理RAW264.7细胞,在处理后3 h和6 h,采用免疫荧光染色法检测MET的产生,并对其进行半定量分析。结果免疫荧光染色表明MET主要由DNA和组蛋白构成,1μg/m L LPS处理RAW264.7细胞6 h,MET形成百分比为(37.04±10.02)%,明显高于对照组(7.90±2.71)%;80μmol/L的PMA处理细胞后MET形成百分比为(22.40±1.83)%,高于对照组(10.11±1.13)%;各剂量的Si O2诱导MET的量与对照组无明显差异。结论用LPS和PMA均能在体外诱导RAW264.7细胞MET的形成,Si O2诱导不是可行的方法。
OBJECTIVE: To compare different methods of inducing macrophage extracellular trapping (MET) formation in vitro. Methods LPS, (10,25,50,80) μmol / L phorbol ester (PMA) and (50,100,150) μg / m RAW264.7 cells were treated with silicon dioxide (Si O2). The production of MET was detected by immunofluorescence staining at 3 h and 6 h after treatment, and the semi-quantitative analysis was performed. Results Immunofluorescence staining showed that MET mainly consisted of DNA and histone. RAW264.7 cells were treated with 1μg / mL LPS for 6 h, the percentage of MET formation was (37.04 ± 10.02)%, which was significantly higher than that of control group (7.90 ± 2.71)%. The percentage of MET formation in cells treated with 80μmol / L PMA was (22.40 ± 1.83)%, higher than that in the control group (10.11 ± 1.13)%. The amount of MET induced by Si O2 was not significantly different from that in the control group. Conclusion Both LPS and PMA can induce MET formation in RAW264.7 cells in vitro. Si O2 induction is not feasible.