广州医药
廣州醫藥
엄주의약
Guangzhou Medical Journal
2015年
5期
5-8
,共4页
梁志科%赵子文%刘朝晖%李裕军%钟维农
樑誌科%趙子文%劉朝暉%李裕軍%鐘維農
량지과%조자문%류조휘%리유군%종유농
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
燐痠二酯酶4抑製劑%咯利普蘭%巨噬細胞%金黃色葡萄毬菌%流感嗜血桿菌
린산이지매4억제제%각리보란%거서세포%금황색포도구균%류감기혈간균
Phosphodiesterases 4 inhibitor%Rolipram%Macrophage%Heamophilus influenzae%Staphylococcus au-reus
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。
目的:探討燐痠二酯酶4抑製劑對人肺泡巨噬細胞(AM)吞噬非生物性顆粒及革蘭暘性菌、陰性菌能力的影響。方法使用 Ficolll-Hypaque 密度梯度法將外週血單覈細胞分離的靜脈血,在含有2 ng/m GM-CSF 的培養液中經12天誘導培養成 AM替代細胞模型—單覈細胞源性巨噬細胞(MDM)。用酶標儀檢測 MDM經燐痠二酯酶4抑製劑 Rolipram 預處理過夜(16~18 h)後吞噬熒光標記的非生物顆粒 Beads 和熱滅活的流感嗜血桿菌(H.influenzae)、金黃色葡萄毬菌(S.aureus)量的改變,另使用 MTT 法檢測細胞活性。結果成功建立的 MDM細胞模型對 Beads 和細菌的吞噬呈時效關。Rolipram 在實驗濃度(10~8~10-5 M)下對 MDM吞噬 Beads、H.influenzae 和S.aureus 能力無明顯促進或抑製作用,也不影響 MDM的活性。結論燐痠二酯酶4抑製劑不會因升高巨噬細胞內cAMP 水平而影響其吞噬非生物顆粒和細菌的能力。
목적:탐토린산이지매4억제제대인폐포거서세포(AM)탄서비생물성과립급혁란양성균、음성균능력적영향。방법사용 Ficolll-Hypaque 밀도제도법장외주혈단핵세포분리적정맥혈,재함유2 ng/m GM-CSF 적배양액중경12천유도배양성 AM체대세포모형—단핵세포원성거서세포(MDM)。용매표의검측 MDM경린산이지매4억제제 Rolipram 예처리과야(16~18 h)후탄서형광표기적비생물과립 Beads 화열멸활적류감기혈간균(H.influenzae)、금황색포도구균(S.aureus)량적개변,령사용 MTT 법검측세포활성。결과성공건립적 MDM세포모형대 Beads 화세균적탄서정시효관。Rolipram 재실험농도(10~8~10-5 M)하대 MDM탄서 Beads、H.influenzae 화S.aureus 능력무명현촉진혹억제작용,야불영향 MDM적활성。결론린산이지매4억제제불회인승고거서세포내cAMP 수평이영향기탄서비생물과립화세균적능력。
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'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.