Objective To investigate the mRNA and protein expression of human β-defensin-2 (hBD-2) induced by lipopolysaccharide (LPS),IL-1β and TNF-α in human airway primary epitheliums.Methods The bronchial primary epitheliums from human were stimulated with LPS,IL-1β and TNF-α respectively and then were harvested for hBD-2 expression detection.The mRNA expression of hBD-2 was detected by RT-PCR,and the protein expression by immunocytochemistry and western blot.Results There was a small expression of hBD-2 mRNA in human airway primary epitheliums before stimulation.The hBD-2 mRNA expression was significantly increased after 3 hours of LPS,IL-1β and TNF-α stimulation respectively and the expression increasement was in a dose dependent manner.The hBD-2 protein could be detected in cytoplasm after 4 hours of LPS (0.1 μg/mL),IL-1β (1 ng/mL) and TNF-α (10 ng/mL) stimulation.Conclusions LPS and proinflammatory cytokines can induce the mRNA and protein expression of hBD-2 in a short time.The expression of hBD-2 may play an initial defense role against bacterial invasion.
目的 研究CD14过表达对胃癌细胞核转录因子-κB(NF-κB)活性以及人β防御素-2(hBD-2)表达的影响,探讨CD14在胃癌发生发展中的作用。 方法 体外培养CD14稳定转染的胃癌SGC-7901细胞系及空质粒转染的对照细胞,CD14蛋白受体胞壁酰二肽刺激细胞,凝胶迁移实验检测NF-κB的活性,蛋白质印迹法检测NF-κB蛋白的表达,同时分别用逆转录-聚合酶链反应以及蛋白质印迹法检测hBD-2 mRNA及蛋白的表达。 结果 与对照组相比,CD14过表达的细胞中NF-κB的活性明显增强,蛋白表达量也大幅度增加,同时hBD-2的mRNA及蛋白的表达都有所提高。 结论 胃癌细胞中CD14在介导NF-κB的激活以及hBD-2的表达中发挥重要作用。