目的研究苯并[a]芘(BaP)作用下热休克转录因子1(HSF1)与热休克元件(HSE)结合特性,分析BaP对热应激蛋白70(HSP70)的表达的影响。方法取对数生长期的猪主动脉内皮细胞传代培养,分别以不同浓度的BaP染毒(0、0.1、0.5、1、5、10μmol/L)24h,提取细胞核蛋白,用凝胶阻滞电泳实验(EMSA)检测结合率。结果低(0.1μM)、中剂量(0.5,1μM)BaP可使HSF1-HSE结合率上升,高剂量(5,10μM)BaP使HSF1-HSE结合率明显降低。结论BaP可改变细胞HSF1-HSE结合能力,低、中剂量的BaP使HSF1-HSE结合率升高,提示可促进HSP70表达,高浓度BaP可抑制HSP70的表达。
Objective To investigate the binding level of heat shock transcriptional factor 1(HSF1) with heat shock element (HSE) of aortic endothelial cells caused by benzo(a)pyrene(BaP) ,and to analyze the effect on HSP70 gene expression of BaP. Methods Porcine aoryie endothelial cells were cultured in vitro, and were treated by different concentration of BaP(0, 0. 1,0. 5 ,1, 5, 10μmol/L) for 24 hours, the binding ability of HSF1 with HSE was further determined by eleetropharetie mobility shift assay (EMSA). Results The binding level of HSF1 to HSE increased at low(0. 1 μmol/L) and medium-exposed group(0. 5,1μmol/L)of BaP, but significantly decreased at high concentration group(5, 10μmol/L). Conclusion The results suggested that the HSP70 gene expression could be increased at low and medium-exposed group of BaP, and inhibited at high concentration group.