氟碳化合物对脂多糖诱导体外培养中性粒细胞NF-κB活化的影响  

Effect of perfluorocarbon on LPS-induced activation of nuclear factor-kkkkkkB in neutrophils in vitro

作　　者：孙震[1] 吕晶[1] 邵素娟 陈卫民[1] 

机构地区：[1]中国医科大学附属第二医院麻醉科,沈阳市110004 [2]辽宁省盘锦市第四人民医院麻醉科

出　　处：《中华麻醉学杂志》2004年第8期612-614,共3页Chinese Journal of Anesthesiology

摘　　要：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 It has been shown that partial liquid ventilation (PLV) with perfluorocarbon (PFC) can reduce intrapulmonary aggregation of neutrophils (PMNs) and macrophages and their cytokine release during acute lung injury (ALI) . The purpose of this study was to investigate the effect of PFC on activation of nuclear factor-kappa B (NF-κB) in neutrophils induced by lipopolysacchride (LPS) .Methods PMNs were isolated from peripheral blood obtained from 36 healthy volunteers and incubated in RPM 1640 cell culture medium yielding a concentration of 1×108/ hole. The PMNs were randomly divided into two groups : (Ⅰ) LPS group in which PMNs were incubated with LPS 10μg·ml-1(n = 18) and (Ⅱ) PFC group in which 5 min after stimulation of PMNs with the same concentration of LPS (10μg·ml-1 ) , PFC was added to the cell suspension with the final concentration of PFC / cell culture medium (vol / vol) of 0.3 (n=18 ). After incubation with LPS for 3 h (T1), 8h (T2) and 24 h (T3), the cell suspensions were centrifuged. The supernatants were analyzed for TNF-α concentration and gray value of activated NF-κB was measured in the nuclear of neutrophils. Results The concentration of TNF-a and gray value of NF-κB in PFC group were significantly lower than those in LPS group at all time points ( P <0.01). The TNF-α concentration and gray value of NF-κB at T1 were significantly lower than those at T2 and T3 in both groups. Conclusion PFC can reduce LPS-induced inflammatory response by inhibiting NF-κB activation in neutrophils.

关 键 词：中性粒细胞 NF-ΚB活化 LPS 脂多糖 体外培养 诱导 TNF-α 导体 结论 显著性 

分 类 号：R96[医药卫生—药理学]