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目的探讨人单核细胞(THP-1)对煤焦沥青烟提取物(CTPE)诱导BEAS-2B细胞肿瘤坏死因子受体相关因子2(TRAF2)mRNA表达的影响。方法构建THP-1细胞和BEAS-2B细胞的共培养模型,用终浓度3μg/m L的CTPE处理共培养模型,细胞培养至第9代时加入肿瘤坏死因子-α(TNF-α)中和抗体和C-Jun氨基端激酶(JNK)抑制剂SP600125,继续培养至第15代(CC15),设立共培养模型CC15组、TNF-α中和抗体组、JNK激酶抑制剂SP600125组,同代培养的BEAS-2B细胞为对照组,用Real-time PCR方法检测各组细胞中TRAF2 mRNA的相对表达水平。结果与CC10组相比,TNF-α中和抗体对共培养细胞作用不同时间时TRAF2 mRNA的相对表达量均降低(P<0.001),作用48 h时TRAF2 mRNA相对表达水平达到最低(0.19±0.02)。SP600125抑制剂作用共培养细胞不同时间时c-Jun mRNA的相对表达水平与CC10组相比差异均有统计学意义(P<0.001),36 h组表达水平最低(0.03±0.02)。TNF-α中和抗体组和SP600125抑制剂组TRAF2 mRNA相对表达水平(分别为1.73±0.04和1.88±0.05)均高于对照组(1.07±0.06),低于CC15组(2.23±0.09),差异有统计学意义(P<0.05)。结论 THP-1可通过TNF-α上调了CTPE诱导的BEAS-2B细胞中TRAF2 mRNA的表达水平,TRAF2和AP-1之间可能存在负反馈作用。
Objective To investigate the effects of human mononuclear cells (THP-1) on the expression of tumor necrosis factor receptor related factor 2 (TRAF2) mRNA in BEAS-2B cells induced by coal tar pitch smoke extract (CTPE). Methods The co-culture model of THP-1 cells and BEAS-2B cells was constructed. The co-culture model was treated with CTPE at a final concentration of 3μg / ml. Neutrophils were treated with tumor necrosis factor-α (CC15), CC15 group, TNF-α neutralizing antibody group and JNK kinase inhibitor SP600125 group were cultured in the same culture Of BEAS-2B cells as control group, Real-time PCR method was used to detect the relative expression of TRAF2 mRNA in each group. Results Compared with CC10 group, the relative expression level of TRAF2 mRNA in the cocultured cells was decreased at different time (P <0.001), and the relative expression level of TRAF2 mRNA reached the lowest at 48 h (0.19 ± 0.02 ). Compared with the CC10 group, the relative expression level of c-Jun mRNA at different time points was significantly different (P <0.001). The expression level of c-Jun mRNA in the 36 h group was the lowest (0.03 ± 0.02). The relative expression levels of TRAF2 mRNA in TNF-α neutralizing antibody group and SP600125 inhibitor group were 1.73 ± 0.04 and 1.88 ± 0.05 respectively, which were significantly lower than those in control group (1.07 ± 0.06) and 2.23 ± 0.09 There was statistical significance (P <0.05). Conclusions THP-1 can up-regulate TRAF2 mRNA expression in CTPE-induced BEAS-2B cells by TNF-α, and there may be a negative feedback between TRAF2 and AP-1.