广西医科大学学报2025，Vol.42Issue(3)：332-341,10.DOI:10.16190/j.cnki.45-1211/r.2025.03.002
铜绿假单胞菌诱导的中性粒细胞胞外诱捕网对树突状细胞的影响
The impact of neutrophil extracellular traps induced by Pseudomonas aeruginosa on den-dritic cells
摘要
Abstract
Objective:To investigate the regulatory effects of neutrophil extracellular traps(NETs)induced by Pseudomonas aeruginosa(PAO1)on dendritic cells(DCs).Methods:Human peripheral blood neutrophils were isolated in vitro and stimulated with phorbol myristate acetate(PMA)and PAO1(at multiplicities of infection,MOI=1,10,100)to induce NET formation.The NET morphology was observed via immunofluorescence stain-ing,and NET-DNA concentration was quantified using PicoGreen fluorescence assay.PAO1-induced NETs were co-cultured with DCs.Flow cytometry was used to detect the expression levels of co-stimulatory molecules(CD40,CD86,CD80)on DC surfaces,while reverse transcription-quantitative polymerase chain reaction(RT-qPCR)and enzyme-linked immunosorbent assay(ELISA)were employed to measure the transcription and secre-tion levels of interleukin-1β(IL-1β),interleukin-12(IL-12),and tumor necrosis factor-α(TNF-α).Results:PAO1-induced NETs exhibited nuclear enlargement and filamentous connection structures.The concentration of NET-DNA in the MOI=100 group was significantly higher than that in the MOI=1 group(P＜0.01).The release of NET-DNA first increased and then decreased from 1 to 3 hours,reaching a peak at 4 hours.PAO1-induced NETs significantly upregulated the expression of co-stimulatory molecules CD40,CD86,and CD80 on the surface of DCs(P＜0.001)and promoted the secretion of IL-1β,IL-12,and TNF-α(P＜0.01).Conclusion:PAO1-induced NETs can promote the maturation and activation of DCs,and induce the secretion of pro-inflammatory cytokines IL-1β,IL-12,and TNF-α.This pro-inflammatory effect may have dual roles.关键词
铜绿假单胞菌/中性粒细胞胞外诱捕网/树突状细胞/免疫调节/细胞因子Key words
Pseudomonas aeruginosa/neutrophil extracellular traps/dendritic cells/immune regulation/cyto-kines分类
医药卫生引用本文复制引用
刘晓彤,罗劲,唐金玲,黄雨婷,杨世毫,孔晋亮..铜绿假单胞菌诱导的中性粒细胞胞外诱捕网对树突状细胞的影响[J].广西医科大学学报,2025,42(3):332-341,10.基金项目
国家自然科学基金资助项目(No.82104499 （）
No.82160783) （）
广西壮族自治区科技厅重点研发计划资助项目(No.桂科AB21196010) （No.桂科AB21196010）
中国博士后科学基金第73批面上资助项目(No.2023MD734158) （No.2023MD734158）
广西自然科学基金青年科学基金项目(No.2023GXNSFBA026146) （No.2023GXNSFBA026146）
