Neutrophils, crucial players in the effector phase of the immune response, are recognized as important mediators of both innate and adaptive immune responses. Through the production of pro- and anti-inflammatory cytok...Neutrophils, crucial players in the effector phase of the immune response, are recognized as important mediators of both innate and adaptive immune responses. Through the production of pro- and anti-inflammatory cytokines, they modulate the function of T and other lymphoid cells. Countless reports have highlighted the importance of these cells as efficient antimicrobial agents and annotated their involvement in the pathology of infectious and noninfectious diseases. The development of modern, sophisticated technologies has allowed the study of the functions of these cells in clinical settings. These advanced technologies include fluorescence-activated cell sorters, confocal microscopy, automated cell image analyzers, and live cell analysis instruments. Unfortunately, the cost of these modern instruments, maintenance, reagents, and the need for qualified technicians prohibit their use in low-income laboratories and universities in developing countries. With this in mind, we propose a series of basic tests that can be used in low-input clinical laboratories and universities to evaluate the function of neutrophils in health and disease. Our methodology allows us to assess in a practical and low-cost manner the functions of neutrophils in the phagocytic process, including opsonization, ingestion, ROI production (NBT reduction), myeloperoxidase content, phagosome-lysosome fusion, microbicidal activity, and NET production. Thus, under a disadvantageous ambiance, this may guide physicians in deciding whether a patient’s illness involves phagocytic defects without imposing a heavy financial burden.Graphical Abstract[-rId13-]展开更多
目的观察康妇炎胶囊对衣原体感染的中性粒细胞(PMN)活性的影响,研究其杀菌消炎机理。方法分离大鼠外周血PMN,衣原体感染6 h后予以10 ng/mL(K-10 ng组)、10 mg/mL(K-10μg组)、1 mg/mL(K-1 mg组)康妇炎胶囊处理6 h、12 h、24 h后,分别收...目的观察康妇炎胶囊对衣原体感染的中性粒细胞(PMN)活性的影响,研究其杀菌消炎机理。方法分离大鼠外周血PMN,衣原体感染6 h后予以10 ng/mL(K-10 ng组)、10 mg/mL(K-10μg组)、1 mg/mL(K-1 mg组)康妇炎胶囊处理6 h、12 h、24 h后,分别收集培养液和细胞分析活性氧簇(ROS)、髓过氧化物酶(MPO)和衣原体载量。利用TLR2/4激动剂或抑制剂,分析康妇炎胶囊影响PMN活性的分子机理。结果康妇炎处理衣原体感染的PMN后,显著降低细胞内ROS(K-1 mg:6 h P=0.000 03、12 h P=0.000 001、24 h P=0.000 003;K-10 mg:6 h P=0.014、12 h P=0.009、24 h P=0.009;K-10 ng:24 h P=0.000 5)和MPO(K-1 mg:6 h P=0.046、12 h P=0.004、24 h P=0.038;K-10μg:12 h P=0.048、24 hP=0.011;K-10 ng:12 h P=0.005、24 h P=0.017)水平,显著降低培养液中衣原体载量(K-10 ng:P=0.000 2;K-10μg:P=0.000 1;K-1 mg:P=0.002)。相比于康妇炎组,TLR4抗体与康妇炎同时作用于细胞,显著抑制PMN分泌MPO(P=0.011);TLR2和TLR4激动剂分别与康妇炎作用细胞,培养液中衣原体负荷量显著降低(PGN:P=0.002;LPS:P=0.000 08)。结论康妇炎胶囊通过抑制PMN产生和分泌ROS和MPO,促进PMN吞噬衣原体,调节PMN的杀菌方式,减少脱颗粒物如ROS、MPO的产生,促进对病原菌的吞噬,减少因PMN脱颗粒释放ROS和MPO到细胞外引起组织损伤,但其调节PMN杀菌方式的分子机理与TLR2/4信号通路不重叠。展开更多
文摘Neutrophils, crucial players in the effector phase of the immune response, are recognized as important mediators of both innate and adaptive immune responses. Through the production of pro- and anti-inflammatory cytokines, they modulate the function of T and other lymphoid cells. Countless reports have highlighted the importance of these cells as efficient antimicrobial agents and annotated their involvement in the pathology of infectious and noninfectious diseases. The development of modern, sophisticated technologies has allowed the study of the functions of these cells in clinical settings. These advanced technologies include fluorescence-activated cell sorters, confocal microscopy, automated cell image analyzers, and live cell analysis instruments. Unfortunately, the cost of these modern instruments, maintenance, reagents, and the need for qualified technicians prohibit their use in low-income laboratories and universities in developing countries. With this in mind, we propose a series of basic tests that can be used in low-input clinical laboratories and universities to evaluate the function of neutrophils in health and disease. Our methodology allows us to assess in a practical and low-cost manner the functions of neutrophils in the phagocytic process, including opsonization, ingestion, ROI production (NBT reduction), myeloperoxidase content, phagosome-lysosome fusion, microbicidal activity, and NET production. Thus, under a disadvantageous ambiance, this may guide physicians in deciding whether a patient’s illness involves phagocytic defects without imposing a heavy financial burden.Graphical Abstract[-rId13-]
文摘目的观察康妇炎胶囊对衣原体感染的中性粒细胞(PMN)活性的影响,研究其杀菌消炎机理。方法分离大鼠外周血PMN,衣原体感染6 h后予以10 ng/mL(K-10 ng组)、10 mg/mL(K-10μg组)、1 mg/mL(K-1 mg组)康妇炎胶囊处理6 h、12 h、24 h后,分别收集培养液和细胞分析活性氧簇(ROS)、髓过氧化物酶(MPO)和衣原体载量。利用TLR2/4激动剂或抑制剂,分析康妇炎胶囊影响PMN活性的分子机理。结果康妇炎处理衣原体感染的PMN后,显著降低细胞内ROS(K-1 mg:6 h P=0.000 03、12 h P=0.000 001、24 h P=0.000 003;K-10 mg:6 h P=0.014、12 h P=0.009、24 h P=0.009;K-10 ng:24 h P=0.000 5)和MPO(K-1 mg:6 h P=0.046、12 h P=0.004、24 h P=0.038;K-10μg:12 h P=0.048、24 hP=0.011;K-10 ng:12 h P=0.005、24 h P=0.017)水平,显著降低培养液中衣原体载量(K-10 ng:P=0.000 2;K-10μg:P=0.000 1;K-1 mg:P=0.002)。相比于康妇炎组,TLR4抗体与康妇炎同时作用于细胞,显著抑制PMN分泌MPO(P=0.011);TLR2和TLR4激动剂分别与康妇炎作用细胞,培养液中衣原体负荷量显著降低(PGN:P=0.002;LPS:P=0.000 08)。结论康妇炎胶囊通过抑制PMN产生和分泌ROS和MPO,促进PMN吞噬衣原体,调节PMN的杀菌方式,减少脱颗粒物如ROS、MPO的产生,促进对病原菌的吞噬,减少因PMN脱颗粒释放ROS和MPO到细胞外引起组织损伤,但其调节PMN杀菌方式的分子机理与TLR2/4信号通路不重叠。
