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目的研究青石棉纤维作用于呼吸道上皮BEAS-2B细胞后白细胞介素-8(IL-8)蛋白和基因的表达水平及酪氨酸激酶抑制剂PD98059对IL-8表达的影响。方法用定量聚合酶链反应法测定青石棉纤维诱导BEAS-2B细胞后的IL-8mRNA水平;用酶联免疫吸附试验(ELISA)法测定BEAS-2B细胞培养上清液中IL-8蛋白水平。结果不同浓度青石棉刺激BEAS-2B细胞后,细胞培养上清液中IL-8释放量明显增加,同样BEAS-2B细胞中IL-8mRNA表达显著上升,使用酪氨酸激酶抑制剂PD98059可明显抑制IL-8的蛋白及mRNA表达水平。结论青石棉可诱导IL-8的高表达,而这种表达可为PD98059所抑制,提示IL-8的表达受丝裂原活化蛋白激酶信号通路中磷酸化ERK蛋白表达的控制。
OBJECTIVE: To study the expression of interleukin-8 (IL-8) protein and gene and the effect of tyrosine kinase inhibitor PD98059 on the expression of IL-8 in cortical epithelial cells of human respiratory tract epithelial cells (BEAS-2B) treated with crocidolite fibers. Methods The level of IL-8 mRNA in BEAS-2B cells induced by crocidolite fibers was determined by quantitative polymerase chain reaction. The level of IL-8 protein in BEAS-2B cell culture supernatants was determined by enzyme linked immunosorbent assay (ELISA). Results After BEA-2B cells were stimulated with crocidolite for different concentrations, the release of IL-8 in cell culture supernatants was significantly increased. Similarly, the expression of IL-8 mRNA in BEAS-2B cells was significantly increased. PD98059, a tyrosine kinase inhibitor, IL-8 protein and mRNA expression levels. Conclusions Crocidolite can induce high expression of IL-8, which can be inhibited by PD98059, suggesting that the expression of IL-8 is under the control of phosphorylated ERK protein in mitogen-activated protein kinase signaling pathway.