【摘要】 目的探讨脂多糖(Lipopolysaccharides,LPS)对人外周血中性粒细胞(neutrophil或polymorphonuclear neutrophil,PMN)吞噬结核分枝杆菌(Mycobacterium tuberculosis,M.tb)功能的影响及Toll样受体4(Toll-like receptor 4,TLR4)在其中发挥的作用。方法取健康成人外周血PMN加入M.tb,抗酸染色、显微镜观察PMN吞噬M.tb情况; PMN与异硫氰酸荧光素(FITC)标记M.tb孵育5～20 min,流式细胞仪检测PMN吞噬率;不同浓度LPS作用PMN,流式细胞术检测PMN吞噬率;应用TLR4单克隆抗体后,流式细胞术检测LPS作用PMN的吞噬率变化。结果 PMN与M.tb作用,经抗酸染色,显微镜油镜观察到中性粒细胞内吞噬有大量粉红色索状M.tb。利用FITC标记M.tb,流式细胞术检测M.tb的标记率在95%。PMN与FITC标记M.tb 37℃水浴作用,利用流式细胞仪检测PMN吞噬率随孵育时间延长而增加。LPS预处理人PMN后,发现PMN 5 min吞噬百分率随LPS浓度增加而增高(P<0.05)。TLR4单克隆抗体作用PMN,再与LPS作用后,PMN吞噬率较未加TLR4单克隆抗体作用有所降低(P<0.05)。结论 LPS可通过TLR4增强PMN吞噬结核分枝杆菌功能。更多还原

【Abstract】 Objective To study the effect of LPS on phagocytosis of Mycobacterium tuberculosis by human neutrophils and the role of TLR4 in this process.Methods The healthy adult peripheral blood was mixed with M.tb,and anti-acid stained was used and the phagocytosis of PMN was observed by microscope. M.tb was labeled with FITC. PMN was incubated with M.tb labeled with FITC for5 to 20 min,the rate of phagocytosis was detected by flow cytometry. PMN was pretreatmented with different concentration of LPS and the rate of phagocytosis was detected by flow cytometry. PMN pretreated with TLR4 monoclonal antibodies was incubated with LPS and the rate of phagocytosis was detected by flow cytometry. Results PMN was observed a lot of pink funicular M.tb in it by oil by acid fast stain and by oil lens of microscope. The labeling rate of M.tb with FITC was about 95% detected by flow cytometry. PMN was incubated with M.tb labeled with FITC for 5 to 20 min. The phagocytosis rate of PMN increasesd with time extending. With pretreatement of different concentration of LPS. The phagocytosis rate increased with the increase of concentration of LPS( P<0.05). With pretreatement of TLR4 monoclonal antibody blocking PMN was incubated with LPS then and with M.tb labeled with FITC. The phagocytosis rate decreased comparing with no blockers( P<0.05). Conclusion LPS can enhance phagocytosis of Mycobacterium tuberculosis by human neutrophils and the mechanism maybe relates to TLR4.更多还原