木材燃烧烟雾颗粒物对人原代呼吸道上皮细胞白细胞介素-8表达水平的影响

摘要

目的 探讨木材燃烧烟雾颗粒物(wood smoke particular,WSP)对人原代呼吸道上皮细胞白细胞介素-8(IL-8)表达水平的影响及机制.方法 采集人呼吸道原代上皮细胞,用0、12.5、25.0、50.0、100.0 μg/ml的WSP刺激人原代呼吸道上皮细胞24 h,用酶联免疫吸附(ELISA)方法检测细胞培养上清中IL-8含量;用表皮生长因子受体(epidermal growth factor receptor,EGFR)特异抑制剂PD153035和p38通路特异抑制剂SB203580分别预处理原代呼吸道上皮细胞,加入100 μg/ml WSP作用24 h,观察其对WSP诱导的IL-8表达水平的影响.结果 WSP作用24 h,各组间IL-8含量差异有统计学意义(P＜0.05).进一步做两两比较,与对照组(0μg/ml)和12.5 μg/ml染毒组比较,25.0、50.0、100.0 μg/ml WSP染毒组原代支气管上皮细胞IL-8含量均明显升高,差异均有统计学意义(P＜0.05).100.0 μg/ml WSP染毒组细胞培养上清中IL-8的含量为(12896.67±1373.11) pg/ml;PD153035+WSP组和SB 203580+WSP组IL-8含量分别为(4546.67±1421.42)和(2803.33±865.00) pg/ml,与WSP组比较,均明显降低,差异均有统计学意义(P＜0.05).结论 WSP可能通过EGFR和p38信号转导途径诱导人原代呼吸道上皮细胞过度表达IL-8.%Objective To investigate the effect and mechanism of interleukin-8 expression induced by wood smoke particles (WSP) in primary human airway epithelial cells.Methods Primary human bronchial epithelial cells (HBEC) were collected through fiberbronchoscopic brushing and incubated with different concentration of WSP (0,12.5,25.0,50.0,100.0 μg/ml).Levels of IL-8 protein in cell culture supernatant were measured using enzyme-linked immune sorbent assay.The specific inhibitors for epidermal growth factor receptor (EGFR) and p38 kinase signaling pathways were employed to pretreat HBEC,respectively,prior to incubation with 100 μg/ml WSP to investigate the mechanism of IL-8 expression induced by WSP.Results The expression of IL-8 was significant increased in a dose-dependent manner after exposure HBEC to different concentration of WSP for 24h.The levels of IL-8 expression were (4546.67±1421.42) pg/ml in (PD153035+WSP) group and (2803.33±865.00) pg/ml in (SB203580+WSP) group respectively,which were significant decreased compared with the level of (12896.67 ± 1373.11) pg/ml in WSP group (P＜0.05).Conclusion WSP could induce IL-8 expression by means of EGFR and p38 signaling pathways.