23价肺炎球菌多糖疫苗体外诱导人气道上皮细胞hBD-2表达的机制

目的探讨23价肺炎球菌多糖疫苗在体外诱导人气道上皮细胞人防御素2（human β-defensin2，hBD-2）表达的机制。方法0．5μg/ml的23价肺炎球茵多糖疫苗刺激原代人气道上皮细胞12h，电泳迁移率变动分析法（electrophoretic mobility shift assay，EMSA）检测原代人气道上皮细胞中NF-KB的活性。激光扫描共聚焦显微镜技术检测原代人气道上皮细胞内钙离子浓度的变化情况。结果23价肺炎球菌多糖疫苗刺激原代人气道上皮细胞24h后．细胞内NF-KB活性增高；静息状态下人气道上皮细胞内钙离子浓度为（38．4±1．21）nmol／L，随着刺激时间的延长，钙离子的浓度逐渐上升，20、30、40min分别为（50．5±3．26）、（51±2．83）、（48±3．47）nmol／L，与对照组相比，差异具有统计学意义（P〈0．05）。结论23价肺炎球菌多糖疫苗诱导hBD-2表达上调是通过激活NF-κB信号通路介导的，细胞内钙离子浓度增高可能是NF-κB激活的重要因素之-。

Expression of hBD-2 induced by 23-valent pneumococcal polysaccharide vaccine in human bronchial epithelial cells

Objective To investigate the mechanisms of up-regulation of human [3-defensin 2 （ hBD- 2） induced by 23-valent pneumococcal polysaccharide vaccine in human bronchial epithelial cells. Methods Primarily cultured human bronchial epithelial cells （HBECs） were stimulated by 23-valent pneumococcal polysaccharide vaccine in vitro. NF-κB activity was evaluated with electrophoretic mobility shift assay （EMSA）. Laser confocal microscopy was conducted to determine intracellular Ca2＋ concentration. Results EMSA showed that the activity of NF-κB was increased 24 h after stimulation by 23-valent pneumococcal polysaccharide vaccine. The intracellular concentrations of Ca2＋ were increased from （38.4 ± 1.21 ） nmol/L to （50.5 ± 3.26）, （51 ± 2.83 ）, （48± 3.47） nmol/L at 20, 30, 40 rain after stimulation, respectively （ P 〈 0.05 ）. Conclusion 23-valent pneumococcal polysaccharide vaccine up-regulates the expression of hBD-2 in HBECs through the activation of NF- κB, which is associated increased intracellular Ca2＋ concentration.