磷酸二酯酶4抑制剂对人巨噬细胞噬菌能力的影响

doi:10.3969/j.issn.1000-8535.2015.05.002

广州医药 ›› 2015, Vol. 46 ›› Issue (5): 5-8.DOI: 10.3969/j.issn.1000-8535.2015.05.002

磷酸二酯酶4抑制剂对人巨噬细胞噬菌能力的影响

梁志科, 赵子文, 刘朝晖, 李裕军, 钟维农

广州市第一人民医院呼吸内科(广州 510180)

收稿日期:2015-03-15 发布日期:2021-11-30
通讯作者: 赵子文,E-mail:zhaozw1963@163.com
基金资助:
广东省自然科学基金(10151006001000015);广东省医学科研基金(A2013495)

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

Liang Zhike, Zhao Ziwen, Liu Zhaohui, et al

Department of Respiratory Medicine,Guangzhou First People's Hospital, Guangzhou 510180,China

Received:2015-03-15 Published:2021-11-30

摘要/Abstract

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。

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

Abstract: Objective To investigate the influence of phosphodiesterases 4 inhibitor on the phagocytosis of non-biological 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. MDM were pretreated with Rolipram overnight (16-18h),phagocytosis of fluorescent labeled beads and H.influenzae,S.aureus by MDM was measured using a fluostar optima fluorimeter. Cell viability was assay with MTT. Results MDM phagocytosis of beads and bacteria was a time-dependant process. Rolipram in the concentration of 10^-8-10^-5M didn't inhibit or promote phagocytosis of beads and bacteria by MDM, and didn't affect the cell viability. Conclusion Phosphodiesterases 4 inhibitor would not affect the human macrophage phagocytic capacity of non-biological particles and bacteria associated with enhanced intracellular cAMP level.

Key words: Phosphodiesterases 4 inhibitor, Rolipram, Macrophage, Heamophilus influenzae, Staphylococcus aureus

梁志科, 赵子文, 刘朝晖, 李裕军, 钟维农. 磷酸二酯酶4抑制剂对人巨噬细胞噬菌能力的影响[J]. 广州医药, 2015, 46(5): 5-8.

Liang Zhike, Zhao Ziwen, Liu Zhaohui, et al. The influence of phosphodiesterases 4 inhibitor on the phagocytosis of bacteria by human macrophage[J]. Guangzhou Medical Journal, 2015, 46(5): 5-8.

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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 macrophage

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