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目的:研究白细胞介素1β(IL-1β)对A549细胞分泌趋化因子白细胞介素8(IL-8)的诱导作用、相关的细胞内信号通道的激活和传导机理。方法:使用IL-1β刺激A549细胞后,Western blot检测细胞内磷酸化ERK1/2(p-ERK1/2),磷酸化p38(p-p38)和磷酸化JNK(p-JNK)蛋白的表达水平;逆转录-聚合酶反应(RT-PCR)检测IL-8 mRNA的表达;ELISA检测IL-8的蛋白水平。结果:IL-1β刺激后细胞内p-ERK1/2、p-p38和p-JNK的蛋白表达明显增加;A549细胞IL-8 mRNA表达明显升高;IL-8的蛋白表达水平明显高于未干预组。MEK1/2激酶抑制物U0126完全阻断了细胞内p-ERK1/2蛋白表达的升高,显著减低了IL-1β诱导的IL-8 mRNA和蛋白表达的增高;p38激酶抑制物SB203580部分阻断了细胞内p-p38表达的增高,对IL-8 mRNA无明显的影响但减低了IL-8蛋白表达的增高。c-Jun氨基末端激酶(JNK)信号通路抑制物SP600125不表现对p-JNK抑制作用,没有影响IL-8mRNA和蛋白的表达。结论:IL-1β通过激活ERK1/2,p38信号通道,介导了A549细胞分泌IL-8。
AIM: To investigate the induction of interleukin-1β (IL-1β) on the secretion of chemokine interleukin-8 (IL-8) by A549 cells and the related activation and transduction mechanisms of intracellular signaling channels. Methods: After A549 cells were stimulated with IL-1β, the expression of phosphorylated ERK1 / 2 (p-ERK1 / 2), phosphorylated p38 (p-p38) and phosphorylated JNK (p-JNK) The expression of IL-8 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) and the protein level of IL-8 by ELISA. Results: The protein expressions of p-ERK1 / 2, p-p38 and p-JNK were significantly increased after IL-1β stimulation; the expression of IL-8 mRNA in A549 cells was significantly increased; the protein expression of IL-8 was significantly higher Intervention group. MEK1 / 2 kinase inhibitor U0126 completely blocked the increase of intracellular expression of p-ERK1 / 2 and significantly decreased the expression of IL-8 mRNA and protein induced by IL-1β; the partial block of p38 kinase inhibitor SB203580 The increase of intracellular p-p38 expression had no significant effect on IL-8 mRNA but decreased the expression of IL-8 protein. The c-Jun N-terminal kinase (JNK) signal pathway inhibitor SP600125 did not show p-JNK inhibition and did not affect IL-8 mRNA and protein expression. Conclusion: IL-1β mediates the secretion of IL-8 by A549 cells through activation of ERK1 / 2 and p38 signaling pathways.