摘要
目的:探讨气道上皮细胞对巨噬细胞表型和吞噬活性的影响及缺氧诱导因子1α(HIF-1α)的作用。方法:将不同浓度(0、100、200、400和800μmol/L)氯化钴(CoCl_2)处理或转染HIF-1αsi RNA的人支气管上皮(HBE)细胞与12-肉豆蔻酸13-乙酸佛波酯(PMA)诱导人单核细胞系THP-1分化的巨噬细胞共培养,RT-qPCR检测HBE细胞HIF-1α的m RNA表达,流式细胞术检测巨噬细胞表面标志物的表达及对大肠杆菌的吞噬率。结果:CoCl_2浓度依赖性增加HBE细胞HIF-1α的m RNA表达,8 h为峰值,同时CoCl_2处理的HBE细胞也增加共培养的巨噬细胞CCL3、CD163、CD206和CCL18荧光强度比率,800μmol/L处理的HBE细胞作用最强;共培养8 h和12 h的巨噬细胞CCL3荧光强度比率增加最明显,而共培养24 h的巨噬细胞CD163、CD206和CCL18荧光强度比率上升更明显。转染HIF-1α-Homo-488 si RNA的HBE细胞对巨噬细胞CCL3、CD163、CD206和CCL18的刺激作用明显减弱。与不同浓度CoCl_2处理的HBE细胞共培养24 h的巨噬细胞对大肠杆菌的吞噬率最初(60 min以前)均呈不同的上升趋势,其后吞噬率均降低。相对于正常HBE细胞共培养组,400和800μmol/L刺激组在各时点的吞噬率均降低,其中800μmol/L刺激组降低最明显。结论:低氧环境下气道上皮细胞早期增强巨噬细胞向M1优势转化,而长期低氧作用的气道上皮细胞抑制巨噬细胞吞噬作用并向M2优势转化,HIF-1α可能是这些过程的重要介质。
AIM: To investigate the effects of airway epithelial cells on the phenotype and phagocytosis of macrophages and the roles of hypoxia-inducible factor-1α( HIF-1α). METHODS: Human bronchial epithelial( HBE) cells treated with CoCl2( 0,100,200,400 and 800 μmol/L) or transfected with HIF-1α si RNA were co-cultured with the macrophages differentiated from human monocyte line THP-1 induced by phorbol 12-myristate 13-acetate( PMA). The m RNA expression of HIF-1α in the HBE cells was detected by RT-qPCR. The expression of macrophage surface markers and the phagocytosis rate of E. coli by macrophages were analyzed by flow cytometry. RESULTS: CoCl2 upregulated the m RNA expression of HIF-1α in the HBE cells in a concentration-dependent manner and peaked at 8 h. HBE cells treated with CoCl2 increased the fluorescence intensity ratio of CCL3,CD163,CD206 and CCL18 in co-cultured macrophages,and the strongest effect was seen in the macrophages co-cultured with HBE cells treated with CoCl2 at 800 μmol/L. The fluorescence intensity ratio of CCL3 in co-cultured macrophages increased most obviously at 8 h and 12 h,while the fluorescence intensity ratio of CD163,CD206 and CCL18 increased more prominently in the macrophages co-cultured for 24 h. The stimulating effects of the HBE cells transfected with HIF-1α-Homo-488 si RNA on CCL3,CD163,CD206 and CCL18 in the macrophages were significantly attenuated. The phagocytosis rate of E. coli by macrophages co-cultured with HBE cells treated with different concentrations of CoCl2 for 24 h initially increased( up to 60 min),and then it gradually decreased. Compared with normal HBE co-culture group,the phagocytosis rate in 400 and 800 μmol/L stimulation groups decreased at each time point,and that in 800 μmol/L stimulation group was the most. CONCLUSION: In hypoxia environment,airway epithelial cells initially transform macrophages predominantly to an M1-phenotype. However,the long-term hypoxia-stimulated airway epithelial cells inhibit the phagocytosis of macrophages and convert t
作者
陈兴无
邢敏
秦立龙
孙珍贵
臧蕾蕾
王寒黎
CHEN Xing-wu;XING Min;QIN Li-long;SUN Zhen-gui;ZANG Lei-lei;WANG Han-li(Department of Respiratory Medicine,The First Affiliated Hospital,Wannan Medical College,Wuhu 241001,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2019年第1期141-149,共9页
Chinese Journal of Pathophysiology
基金
安徽省自然科学基金资助项目(No.1608085MH192)
弋矶山医院引进人才科研基金资助项目(No.YR201401)
关键词
气道上皮细胞
巨噬细胞
表型
吞噬
缺氧诱导因子1Α
Airway epithelial cells
Macrophages
Phenotype
Phagocytosis
Hypoxia-inducible factor-1α
