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目的研究氟碳化合物(PFC)对脂多糖(LPS)诱导体外培养中性粒细胞NF-κB活化的影响。方法将36份中性粒细胞(每份均由200 ml健康成年人全血中提取而得)置于每孔预先放人1.5ml含10％胎牛血清的RPMI 1640培养液的细胞培养板内进行原代培养,调节中性粒细胞数为1×108/孔。随机分成两组,每组18个孔。LPS组(培养液中LPS的终浓度为10μg/ml)和PFC组(培养液中LPS的终浓度为10μml/37℃培养5 min后加入PFC 450μl)。以LPS刺激后第3、8、20小时为研究时点,分别测量细胞培养上清液中肿瘤坏死因子-α(TNF-α)浓度和中性粒细胞NF-κB活化灰度值。结果PFC组细胞培养上清液中TNF-α的浓度低于LPS组(P<0.01),NF-κB活化灰度值也低于LPS组(P<0.01);两组LPS刺激后第3小时的上清液中TNF-α的浓度和NF-κB活化灰度值也低于第8、20小时,而LPS刺激后第8、20小时之间差异无显著性。结论PFC可能是通过抑制中性粒细胞NF-κB的活化来减少炎性因子的释放。
Objective To investigate the effect of fluorocarbon (PFC) on the activation of NF-κB induced by lipopolysaccharide (LPS) in cultured neutrophils in vitro. Methods Thirty-six neutrophils, each extracted from whole body blood of 200 ml healthy adults, were placed in a cell culture plate in which 1.5 ml of RPMI 1640 medium containing 10% fetal bovine serum was placed in each well Primary culture was carried out to adjust the number of neutrophils to 1 × 10 8 / well. Randomly divided into two groups of 18 holes each. LPS group (the final concentration of LPS in the culture medium was 10μg / ml) and the PFC group (the final concentration of LPS in the culture medium was 10μml / 5min after the incubation at 37 ℃). The concentrations of tumor necrosis factor-α (TNF-α) and neutrophil NF-κB activation in the cell culture supernatants were measured at 3, 8 and 20 hours after LPS stimulation. Results The concentration of TNF-α in cell culture supernatant of PFC group was lower than that of LPS group (P <0.01), and the activation gray value of NF-κB was also lower than that of LPS group (P <0.01) The concentration of TNF-α in supernatant and the activation gray value of NF-κB were also lower than the 8th and 20th hours, but there was no significant difference between the 8th and 20th hour after LPS stimulation. Conclusion PFC may reduce the release of inflammatory cytokines by inhibiting the activation of NF-κB in neutrophils.