人单核细胞对煤焦沥青烟提取物诱导BEAS-2B细胞TRAF2 mRNA表达的影响

Effect of human monocytes on coal tar pitch extract-induced TRAF2 mRNA expression in human bronchial epithelial cells

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 explore the regulation mechanism of coal tar pitch smoke extracts （CTPE）on tumor necrosis factor receptor-associated factor 2 （TRAF2） mRNA expression in co-cultured model of human monocytes （THP-1）and BEAS-2B cells. Methods The co-cultured THP-1 and BEAS-2B cells were simulated by 3 μg/mL CTPE. The ceils at passage 9 were treated with tumor necrosis factor-α （TNF-α） neutralization antibody and c-Jun N-terminal kinase （JNK） inhibitors SP600125, and continued to culture at passage 15 （neutralizing antibody group） and established the BEAS-2B cells of same generation as control group. The expression of TRAF2 mRNA was detected by real- time fluorescent quantitative polymerase chain reaction. Results The expression of TRAF2 mRNA and c-Jun mRNA in different groups of TNF-α neutralization antibody and JNK kinase inhibitors SP600125 groups were decreased compared with CC10, respectively the groups of 48h （0.19 ±0.02） and 36h （0.03 ±0.02） were lowest. The expression of TRAF2 mRNA in groups of TNF-α neutralization antibody and JNK kinase inhibitors SP600125 groups （ respectively 1.73 ± 0.04 and 1.88 ± 0.05 ） were both significantly increased compared with control group （ 1.07 ± 0.06） , and lower than CC15 （2.23 ± 0.09）. Conclusion THP-1 could up-regulate the expression of TRAF2 mRNA through TNF-α in the BEAS-2B cells stimulated by CTPE , and there maybe negative feedback effect between TRAF2 and AP-1.