摘要
目的:检验b型流感嗜血杆菌结合疫苗在体外能否刺激人气道上皮细胞hBD-2表达,并探讨人工诱导防御素表达的方法。方法:体外分离、培养原代人气道上皮细胞。b型流感嗜血杆菌疫苗刺激原代人气道上皮细胞后,用RT-PCR和ELISA法检测培养上清液中hBD-2蛋白的表达。收集刺激后的细胞培养上清液进行抑菌实验。结果:浓度>1μg/ml的b型流感嗜血杆菌结合疫苗刺激原代人气道上皮细胞12 h后,可诱导hBD-2 mRNA的表达,与对照组相比较有显著差异(P<0.01),无剂量依赖性,同时细胞培养上清液中hBD-2蛋白的表达上调。1μg/ml b型流感嗜血杆菌结合疫苗浓缩后的细胞培养上清液具有抑菌作用。结论:一定浓度(1μg/mL)以上的b型流感嗜血杆菌结合疫苗作用于人原代气道上皮细胞可诱导hBD-2的表达,为人工方法刺激机体产生人β防御素-2,对抗病原菌感染提供新的策略。
Objective： This study was undertaken to evaluate if the expression of hBD-2 could be enforced by the stimulation of type b hemophilus influenzae binding vaccine in human bronchial epithelial cells and explore the methods that could induce the expression of hBD-2.Methods： The human bronchial epithelial cells were isolated,cultured and then stimulated by different concentration of type b hemophilus influenzae binding vaccine.RT-PCR and ELISA were performed to determine the expression of hBD-2 mRNA and protein respectively.The supernatant of the cell culture was collected after the stimulation to implement the bacteriostasis experiment.Results： Type b hemophilus influenzae binding vaccine at the concentration above 1 μg/ml could increase the expression of hBD-2 mRNA and protein expression in the human bronchial epithelial cells 12 hours after the stimulation,（P 0.01）,but there was no dosage-dependence.The condensed supernatant acquired the ability of bacteriostasis.Conclusion： Type b hemophilus influenzae binding vaccine greater than 1 μg/ml significantly increases the expression of hBD-2mRNA and protein,providing a new approach of inducing the hBD-2 expression for antimicrobial treatment.
出处
《现代生物医学进展》
CAS
2013年第5期804-807,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(30670932)
