IL-1β诱导支气管上皮细胞高迁移率族蛋白1主动释放

发布时间：2019-01-16 01:57

【摘要】：目的研究IL-1β对健康人支气管上皮细胞(HBE)高迁移率族蛋白1(HMGB1)表达和释放的影响。方法培养HBE135-E6E7,四唑盐(MTT)法检测不同浓度IL-1β对支气管上皮细胞活力的影响,荧光定量PCR检测IL-1β刺激HBE后HMGB1mRNA表达,双抗体夹心ELISA法检测细胞上清液中HMGB1水平,蛋白免疫印迹方法检测IL-1β刺激HBE总蛋白、胞核、胞质HMGB1表达。免疫荧光观察IL-1β刺激后对HBE HMGB1的移位的影响。结果 0.1、1.0、10.0ng/mL IL-1β刺激HBE 24h对其活力无影响。IL-1β以浓度和时间依赖性方式刺激HBE的HMGB1mRNA水平升高,IL-1β增加HBE的HMGB1蛋白表达水平。在浓度依赖性实验,与对照组比较,1.0、10.0ng/mL IL-1β刺激HBE 24h后培养上清液HMGB1水平显著增加;在时间依赖性实验中,与对照组比较,10.0ng/mL IL-1β刺激HBE 12h、24h后细胞上清液中的HMGB1显著升高(P0.05);10.0ng/mL IL-1β刺激HBE 24h后,HBE胞质蛋白明显增加,胞核蛋白减少。免疫荧光检测显示,HMGB1主要表达正常HBE的细胞核内,少量分布于细胞质,IL-1β(10.0ng/mL)刺激HBE 24h,HMGB1明显从HBE胞核向胞质转位。结论 IL-1β可以显著诱导支气管上皮细胞HMGB1的表达和主动释放,提示HMGB1可能参与了IL-1β介导慢性气道炎症过程。
[Abstract]:Objective to study the effect of IL-1 尾 on the expression and release of (HBE) high mobility group protein 1 (HMGB1) in human bronchial epithelial cells. Methods HBE135-E6E7, tetrazolium (MTT) assay was used to detect the effect of different concentrations of IL-1 尾 on the viability of bronchial epithelial cells. The expression of HMGB1mRNA in HBE stimulated by IL-1 尾 was detected by fluorescence quantitative PCR, and the level of HMGB1 in supernatant of cells was detected by double antibody ELISA method. The expression of total protein, nucleus and cytoplasm HMGB1 of HBE stimulated by IL-1 尾 was detected by Western blot. The effects of IL-1 尾 stimulation on HBE HMGB1 translocation were observed by immunofluorescence. Results 0.1 mg / mL IL-1 尾 had no effect on the activity of HBE. IL-1 尾 increased the HMGB1mRNA level of HBE in a concentration-and time-dependent manner, and IL-1 尾 increased the HMGB1 protein expression of HBE. In the concentration-dependent experiment, compared with the control group, the level of HMGB1 in the supernatant of HBE stimulated by 1.0 渭 g / mL IL-1 尾 for 24 h was significantly increased. In the time-dependent experiment, compared with the control group, the HMGB1 in the supernatant of 10.0ng/mL IL-1 尾 increased significantly after HBE was stimulated for 12 h or 24 h (P0.05). After 10.0ng/mL IL-1 尾 stimulated HBE for 24 h, HBE cytoplasmic protein increased significantly and nuclear protein decreased. Immunofluorescence assay showed that HMGB1 mainly expressed normal HBE in the nucleus and a small amount was distributed in the cytoplasm. IL-1 尾 (10.0ng/mL) stimulated HMGB1 translocation from HBE nucleus to cytoplasm for 24 h. Conclusion IL-1 尾 can significantly induce the expression and active release of HMGB1 in bronchial epithelial cells, suggesting that HMGB1 may be involved in the process of chronic airway inflammation mediated by IL-1 尾.
【作者单位】：广西医科大学第一附属医院西院呼吸内科;
【基金】：广西医科大学青年基金资助项目(GXMUSF201206)
【分类号】：R562.25

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