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目的探讨二氧化硅(SiO2)刺激矽肺患者肺泡巨噬细胞(AM)条件培养上清对人胚肺成纤维细胞(FB)白细胞介素(IL)-1β表达的影响。方法用酶联免疫吸附法检测离体经或不经SiO2刺激的矽肺患者AM在3、6、12、18、24、36 h上清中IL-1β的表达,取IL-1β表达高峰点的AM上清作为条件培养上清。采用免疫细胞化学法检测AM条件培养上清刺激FB于不同时间点(3、6、12、18、24、36、48 h)其细胞内IL-1β的表达。结果经SiO2刺激的矽肺患者AM上清中IL-1β的表达量明显升高(P<0.01),12 h达高峰;活化的矽肺患者12 h AM条件上清可以促进FB表达IL-1β增高(P<0.05);18 h表达最高,以后逐渐下降。结论在本实验条件下,经离体细胞培养,提示SiO2可通过上调AM和FB中IL-1β的表达,参与矽肺纤维化的形成过程。
Objective To investigate the effect of conditioned media of conditioned medium of alveolar macrophages (AM) on the expression of interleukin (IL) -1β in human embryonic lung fibroblasts (FB) stimulated by silica (SiO2). Methods The levels of IL-1β in supernatants of AM were detected by enzyme-linked immunosorbent assay (AMI) at 3, 6, 12, 18, 24, 36 h with or without SiO2 stimulation. The peak of IL-1β expression AM supernatant as a condition culture supernatant. Immunocytochemistry was used to detect the expression of IL-1β in FB cells stimulated with AM conditioned medium at different time points (3,6,12,18,24,36,48 h). Results The level of IL-1β in AM supernatant of patients with silicosis stimulated by SiO2 was significantly increased (P <0.01), and peaked at 12 hours. The activated supernatant of silicosis patients with 12-h AM increased the expression of IL-1β in FB P <0.05). The expression was highest at 18 h and then decreased gradually. Conclusion Under the conditions of this experiment, it was suggested that SiO2 could participate in the formation of silicotic fibrosis by up-regulating the expression of IL-1β in AM and FB.