磷酸二酯酶4抑制剂对人巨噬细胞噬菌能力的影响  

The influence of phosphodiesterases 4 inhibitor on the phagocytosis of bacteria by human macrophages

作　　者：梁志科[1] 赵子文[1] 刘朝晖[1] 李裕军[1] 钟维农[1] 

机构地区：[1]广州市第一人民医院呼吸内科,广州510180

出　　处：《广州医药》2015年第5期5-8,共4页Guangzhou Medical Journal

基　　金：广东省自然科学基金(10151006001000015);广东省医学科研基金(A2013495)

摘　　要：目的：探讨磷酸二酯酶4抑制剂对人肺泡巨噬细胞（AM）吞噬非生物性颗粒及革兰阳性菌、阴性菌能力的影响。方法使用 Ficolll-Hypaque 密度梯度法将外周血单核细胞分离的静脉血，在含有2 ng／m GM-CSF 的培养液中经12天诱导培养成 AM替代细胞模型—单核细胞源性巨噬细胞（MDM）。用酶标仪检测 MDM经磷酸二酯酶4抑制剂 Rolipram 预处理过夜（16～18 h）后吞噬荧光标记的非生物颗粒 Beads 和热灭活的流感嗜血杆菌（H.influenzae）、金黄色葡萄球菌（S.aureus）量的改变，另使用 MTT 法检测细胞活性。结果成功建立的 MDM细胞模型对 Beads 和细菌的吞噬呈时效关。Rolipram 在实验浓度（10～8～10-5 M）下对 MDM吞噬 Beads、H.influenzae 和S.aureus 能力无明显促进或抑制作用，也不影响 MDM的活性。结论磷酸二酯酶4抑制剂不会因升高巨噬细胞内cAMP 水平而影响其吞噬非生物颗粒和细菌的能力。Objective To investigate the influence of phosphodiesterases 4 inhibitor on the phagocytosis of non-biologi-cal particles and gram-positive bacteria,gram-negative bacteria by human alveolar macrophages. Methods Peripheral blood mononuclear cells （PBMCs）were isolated from venous blood from 12 healthy volunteers using Ficoll-Hypaque density gradients.Monocytes were incubated with media containing 2 ng/ml GM-CSF for 12d to allow full differentiation into macrophage （MDM）, a functionally equivalent model of human AM.MDMwere pretreated with Rolipram overnight （16-18h）,phagocytosis of fluores-cent labeled beads and H.influenzae,S.aureus by MDMwas measured using a fluostar optima fluorimeter.Cell viability was as-say with MTT. Results MDMphagocytosis of beads and bacteria was a time-dependant process.Rolipram in the concentration of 10-8-10-5Mdidn＆#39;t inhibit or promote phagocytosis of beads and bacteria by MDM,and didn＇t affect the cell viability. Conclu-sion Phosphodiesterases 4 inhibitor would not affect the human macrophage phagocytic capacity of non-biological particles and bacteria associated with enhanced intracellular cAMP level.

关 键 词：磷酸二酯酶4抑制剂 咯利普兰 巨噬细胞 金黄色葡萄球菌 流感嗜血杆菌 

分 类 号：R96[医药卫生—药理学]