煤焦沥青烟提取物对 BEAS-2 B细胞环氧化酶-2 mRNA 表达的影响

摘要

To examine the effect of coal tar pitch smoke extract (CTPE) on COX-2 gene expression in human bronchial epithelial cells .Methods:The human bronchial epithelial cell line ( BEAS-2B) was used as an in vitro model . CTPE induced COX-2 mRNA expression were measured by RT-PCR.Actinomycin D,a transcription inhibitor, was applied to test the effect of CTPE on COX-2 gene transcription .mRNA decay assay was used to observe the effect of CTPE on COX-2 mRNA stability at post transcriptional level .Results:CTPE stimulation increased COX-2 mRNA expression in BEAS-2B cells(Fgroup =71.750,Ftime =93.120,Finteraction =17.300,all P<0.001).Pretreatment of BEAS-2B cells with Actinomycin D (1 mg/L),for 30 min could significantly reduce the effect of CTPE on COX-2 mRNA expression (FAct D =174.203, FCTPE =260.312,Finteraction =188.521,P<0.001).Actinomycin D was used to terminate COX-2 mRNA synthesis elevated by zinc sulfate.At different time points (0, 2, 4 h) COX-2 mRNA levels were higher in CTPE group than those in control group(Fgroup =365.700,Ftime =37.780,Finteraction =6.240,all P<0.05).Conclusion: CTPE stimulation can significantly increase COX-2 mRNA expression levels in BEAS-2B cells.%目的：探讨煤焦沥青烟提取物（ CTPE）对人支气管上皮细胞环氧化酶-2（ COX-2） mRNA表达的影响。方法：应用RT-PCR方法测定不同质量浓度（2、4、8 mg/L）CTPE作用不同时间（2、4、8 h）对人支气管上皮BEAS-2B细胞COX-2 mRNA表达水平影响。使用放线菌素（Act）D（1 mg/L）作用于BEAS-2B细胞，观察其对2 mg/L CTPE作用30 min后的BEAS-2B细胞COX-2基因转录的影响。利用ZnSO4溶液刺激BEAS-2B细胞COX-2 mRNA表达，去除ZnSO4后，加入1 mg/L的Act D 30 min，检测CTPE 作用不同时间点（0、2、4 h）的COX-2 mRNA表达的变化。结果：2、4、8 mg/L CTPE刺激均可以使BEAS-2B细胞COX-2 mRNA表达水平升高（ F组间＝71．750，F时间＝93．120， F交互＝17．300，P＜0．001），2 mg/L是最佳剂量。用Act D（1 mg/L）预处理细胞30 min，可降低CTPE对COX-2 mR-NA表达的诱导作用（FAct D ＝174．203，FCTPE ＝260．312，F交互＝188．521，P＜0．001）。使用ZnSO4刺激，然后使用Act D终止COX-2 mRNA合成后，不同时间点， CTPE 组和对照组COX-2 mRNA的表达差异有统计学意义（ F组间＝365．700，F时间＝37．780， F交互＝6．240，P＜0．001）。结论：CTPE可以通过转录水平和转录后水平诱导人支气管上皮细胞COX-2 mRNA的表达。