IL-1β/p38MAPK/NF-κB信号转导通路体外调节鼻黏膜上皮细胞中糖皮质激素受体的表达

The involvement of IL-1β/p38MAPK / NF-κB signal transduction pathway in regulating the expression of glucocorticoid receptor isoformsin human nasal epithelia in vitro

目的:拟通过体外研究初步探讨慢性鼻-鼻窦炎黏膜中GRα/GRβ降低的可能上游信号转导机制。方法:以IL-1β体外诱导人鼻黏膜上皮细胞(HNE)构建GRα/GRβ降低的模型。以Western印记及荧光定量PCR技术检测IL-1β诱导前后及p38MAPK和NF-κB信号通路特异性阻断剂干预前后GRα、GRβ和p38MAPK、NF-κB信号通路关键酶在蛋白质及核酸水平的表达变化。结果:IL-1β呈浓度和时间依赖性诱导HNE中GRα/GRβ降低,成功建立GRα/GRβ降低的细胞模型。相同梯度的IL-1β可呈相同规律诱导HNE中p38MAPK和NF-κB通路激活。p38MAPK和NF-κB通路的特异性阻断剂SB203580和吡咯烷二硫代氨基甲酸(PDTC)均可明显抑制各自通路,且同时出现GRα/GRβ增高。SB203580可明显降低NF-κBp50和NF-κBp65的表达,PDTC对p38MAPK的表达未见明显影响。结论:IL-1β通过先后激活p38MAPK和NF-κB信号通路诱导体外培养的HNE中GRα/GRβ降低。

Objective To explore the possible upstream signal transduction mechanisms responsible for the decrease of glucocorticoid receptor（GR）α/GRβ in chronic rhinosinusitis（CRS） in vitro.Methods The GRa/GRβ decrease cell model was established by IL-1β-induced human nasal epithelia（HNE） in vitro.Changes in the protein and mRNA expression of GRα,GRβ and the key enzymes in the p38 MAPK and NF-κB pathways were measured respectively,before and after being induced with different doses of IL-1β and specific inhibitors of p38 MAPK and NF-κB.Analysis of variance（ANOVA） was used to analyze the data.Results With IL-1β-induction,the GRα/GRβ ratio declined in both a time-dependent manner and a concentration-dependent manner in HNE,which demonstrated the successful establishment of a GRα/GRβ decrease model in vitro.After cultured HNE were induced with the same set of IL-1β,the p38 MAPK and NF-κB signal pathways were also activated.Either a specific inhibitor（SB203580） of the p38 MAPK pathway or a specific inhibitor（PDTC） of the NF-κB pathway increased the GRα/GRβ ratio at the meantime of inhibiting their pathways.The expressions of phosphoNF-κBp50 and phospho-NF-κBp65 were significantly decreased by SB203580.PDTC had no influence on the expression of the key enzymes in the p38 MAPK.Conclusion The decrease of GRα/GRβ ratio in HNE induced by IL-1β in vitro is induced through the p38MAPK/NF-κB signal pathways.