23价肺炎球菌多糖疫苗刺激人气道上皮细胞hBD-2表达的研究  

Study of expression of hBD-2 in human epithelia cells stimulated by 23 pneumococcal polysaccharide vaccine

作　　者：沈桢巍[1] 蒋易[1] 雷撼[2] 

机构地区：[1]同济大学附属东方医院重症监护室,上海200120 [2]同济大学附属东方医院呼吸内科,上海200120

出　　处：《国际呼吸杂志》2013年第6期407-411,共5页International Journal of Respiration

基　　金：国家自然基金资助项目（30670932）

摘　　要：目的研究以23价肺炎球菌多糖疫苗刺激人原代气道上皮细胞后能否促进8防御素2（hBD-2）的表达及分泌，探讨人工诱导增强防御素表达的方法。方法分离培养人原代气道上皮细胞，以不同浓度23价肺炎球菌多糖疫苗刺激人原代气道上皮细胞，分别于6、12、24h提取细胞总RNA及培养细胞上清液，R，rPCR、ELISA检测hBD-2的mRNA和蛋白表达。纸片法观察杀菌效果。结果不同浓度23价肺炎球菌多糖疫苗刺激气道上皮细胞，6h后各组细胞中hBD-2的mRNA和蛋白表达与对照组相比无明显变化；12h后，疫苗浓度〉0．5mg／L的各组细胞中mRNA表达和蛋白水平显著升高（P〈0．05），但24h后均下降。疫苗（0．5mg／L）作用后12h的细胞培养上清有明显的杀菌效果。结论一定浓度的23价肺炎球菌多糖疫苗（〉0．5mg／L）刺激人原代气道上皮细胞，可促进hBn2mRNA的表达和hBD-2蛋白浓度的增加，从而发挥hBD-2天然抗微生物的作用。Objective To study if the expression of hBD-2 in human epithelia cells could be promoted after stimulated by 23 pneumococcal polysaccharide vaccine, and to explore the method that could induce the expression of hBD-2. Methods Human original airway epithelia cells were isolated and cultured, stimulated by different concentrations of 23 pneumocoecal polysaccharide vaccine. The total RNA in cells and supernatant were collected after 6,12,24 hours, and the expressions of hBD-2 mRNA and protein were respectively detected with RT-PCR and ELISA. Results After stimulated for 6 hours, there were no significant changes in expressions of hBD-2 mRNA and protein between stimulated groups and control group. But after stimulated for 12 hours, the expressions of hBD-2 mRNA and protein in supernatant increased in the groups that the concentration of 23 pneumococcal polysaccharide vaccine over 0.5 mg/L co,rr/pared with control group （ P d0.05）. After 24 hours, the expressions of hBD-2 mRNA and protein in supernatant decreased. Conclusions 23 pneumococcal polysaccharide vaccine above some certain concentration （〉 0.5 rag/L） can stimulate cultured human airway epithelial cells, increase the expressions of hBD-2 mRNA and protein, and hBD-2 plays the role of natural antimicrobial activity.

关 键 词：23价肺炎球菌多糖疫苗 人原代气道上皮细胞 β防御素2 

分 类 号：R392[医药卫生—免疫学]