JNK信号通路在白介素-1β介导人胚肺成纤维细胞活化中的作用

Role of c-Jun N-terminal Kinase Signal Pathway on Interleukin-1β Induced Activation of Human Embryonic Lung Fibroblast

目的探讨c-Jun氨基末端激酶(JNK)信号转导通路在白细胞介素-1β(IL-1β)介导的人胚肺成纤维细胞(FB)活化中的作用。方法体外培养人胚肺FB,用IL-1β、JNK抑制剂(SP600125)作用于细胞,采用免疫蛋白印迹法(Western blot)检测细胞JNK/SAPK磷酸化水平和α-平滑肌激动蛋白(α-SMA)的表达,RT-PCR方法检测纤溶酶原激活物抑制剂(PAI-1)、纤维连接蛋白(FN)、神经生长因子(NGF)和α-SMA mRNA的相对表达量,ELISA法检测培养上清液中NGF及白细胞介素-6(IL-6)水平。结果在IL-1β刺激下,人胚肺FB中α-SMA及磷酸化JNK蛋白表达显著高于对照组,同时PAI-1、FN、NGF和α-SMA mRNA的表达及培养上清液中NGF、IL-6水平均明显高于对照组(P<0.05);SP600125显著抑制IL-1β诱导的上述指标表达增加(P<0.05)。结论 IL-1β促进人胚肺FB活化及细胞外基质聚集,JNK信号转导通路参与这一过程。

Objective To explore the role of c-Jun N-terminal kinase （JNK） signal pathway on interleukin-1β （IL-1β） induced activation of human embryonic lung fibroblasts （FB）.Methods The human embryonic lung fibroblasts were cultured in vitro and treated with IL-βor JNK inhibitor SP600125. The expression of phosphorylated JNK （p-JNK） and α-smooth muscle actin （α-SMA） were examined by Western blot, and the plasminogen activator inhibitor of endothelial cell type （PAI-1）, fibrinogen （FN）, nerve growth factor （NGF） and α-SMA mRNA were detected by RT-PCR. In addition, the concentrations of NGF and interleukin-6 （IL-6） levels in the culture supernatant were measured by enzyme-linked immunosorbent assay （ELISA）.Results We found that under the treatment of IL-1β, the JNK signal pathway in human lung fibroblast was activated. The expression of p-JNK increased significantly compared with the control group （P〈0.05）. mRNA expressions of PAI-1, FN,NGF, α-SMA, and the concentrations of NGF and IL-6 in the culture supernatant were all increased in the IL-1β treated groups （P〈0.05）. However, the SP600125 reversed the effect of IL-1β on the expression of p-JNK, α-SMA, FN, NGF, and IL-6 mentioned above.Conclusion These findings provided evidence that JNK signal pathway may play an important role in the IL-1β induced human lung fibroblast activation and extracellular matrix synthesis.