多形核中性粒细胞在铜绿假单胞菌气溶胶肺递送感染小鼠急性肺损伤中的作用研究  

The role of polymorphonuclear neutrophils in the process of acute lung injury in mice infected via aerosolized intratracheal inoculation of Pseudomonas aeruginosa

作　　者：肖楠 张志鹏 李莎 邓梦芸 王一风 宗福良 苏铎 周冬生 杨慧盈 XIAO Nan;ZHANG Zhi-peng;LI Sha;DENG Meng-yun;WANG Yi-feng;ZONG Fu-liang;SU Duo;ZHOU Dong-sheng;YANG Hui-ying(College of Life Sciences and Technology,Beijing University of Chemical Technology,Beijing 100020,China;State Key Laboratory of Pathogen and Biosecurity,Institute of Microbiology and Epidemiology,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100071,China)

机构地区：[1]北京化工大学生命科学与技术学院,北京100020 [2]军事科学院军事医学研究院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071

出　　处：《军事医学》2022年第10期749-757,共9页Military Medical Sciences

摘　　要：目的 探究多形核中性粒细胞(PMN)在铜绿假单胞菌(PA)气溶胶肺递送感染小鼠急性肺损伤(ALI)过程中的作用。方法 腹腔注射Gr-1抗体构建PMN耗竭小鼠模型,采用流式细胞术和免疫荧光检测PMN的耗竭效果。感染后观察小鼠存活状况,评价肺病理程度,检测肺细菌载量和肺泡灌洗液中细胞因子浓度变化,探究PMN在ALI过程中的作用。结果 流式及免疫荧光结果显示,PMN耗竭小鼠模型构建成功。与对照组小鼠相比,PMN耗竭组小鼠感染后死亡率显著增加,肺部细菌载量增多;肺中仅有少量炎性细胞浸润,肺泡腔中可见大量细菌;肺泡灌洗液中白细胞介素-6(IL-6)、IL-1β、IL-18、IL-17A、IL-22和肿瘤坏死因子-α(TNF-α)等促炎因子表达上调,单核细胞趋化蛋白-1(MCP-1)、巨噬细胞炎性蛋白-2(MIP-2)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)等趋化因子表达上调,IL-4、IL-10、IL-13等抑炎因子表达上调。结论 在感染过程中,PMN的募集有效抑制了细菌繁殖,PMN耗竭后小鼠多种细胞因子代偿性升高,PMN在ALI过程中可能抑制炎症过度发展,是炎症和免疫反应的重要调节剂。Objective To explore the role of polymorphonuclear neutrophils(PMN)in acute lung injury(ALI)in mice infected by aerosolized intratraheal inoculation of Pseudomonas aeruginosa(PA). Methods A mouse model of depleted PMN was constructed via intraperitoneal injection of anti-Gr-1 antibody,while PMN depletion efficiency was assessed by flow-cytometry and immunofluorescence. The survival of mice was observed after infection,the pathological degree of lungs was evaluated,and the changes in bacterial loads of lungs and cytokine concentration in alveolar lavage fluid were detected to analyze the role of PMN in the process of ALI. Results Flow cytometry and immuno-fluorescence results showed that a mouse model of depleted PMN was constructed. Compared with control mice,the mortality and lung bacterial loads of PMN-depleted mice were increased after infection. There was only a small amount of inflammatory cell infiltration in the lung but a large number of bacteria were visible in the alveolar lumen. The expressions of pro-inflammatory factors such as interleukins-6(IL-6),IL-1β,IL-18,IL-17A,IL-22 and tumor necrosis factor-α(TNF-α)were upregulated,so were the expressions of chemokines such as monocyte chemotactic protein-1(MCP-1),macrophage inflammatory protein-2(MIP-2)and granulocyte-macrophage colony-stimulating factor(GM-CSF). The expressions of anti-inflammatory factors such as IL-4,IL-10 and IL-13 were also upregulated in alveolar lavage fluid. Conclusion The recruitment of PMN effectively inhibits the colonization of bacteria during infection. The compensatory elevation of multiple cytokines in mice after PMN depletion suggests that PMN can inhibit the excessive development of inflammation during ALI and PMN can be an important regulator of inflammatory and immune responses.

关 键 词：铜绿假单胞菌 多形核中性粒细胞 气溶胶肺递送 急性肺损伤 

分 类 号：R563.1[医药卫生—呼吸系统]