中性粒细胞在蓖麻毒素肺中毒小鼠肺脏炎症发展中的作用研究  

Study on the role of neutrophils in the development of pulmonary inflammation in mice with ricin poisoning

作　　者：李莎 焦周光 苏铎 干长姣 肖楠 邓梦芸 周冬生 杨慧盈 高波 LI Sha;JIAO Zhou-guang;SU Duo;GAN Chang-jiao;XIAO Nan;DENG Meng-yu;ZHOU Dong-sheng;YANG Hui-ying;GAO Bo(Anhui Medical University,Hefei 230000,China;State Key Laboratory of Pathogen and Biosecurity,Institute ofMicrobiology and Epidemiology,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100071,China)

机构地区：[1]安徽医科大学,合肥230000 [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》2021年第6期426-430,442,共6页Military Medical Sciences

摘　　要：目的探索中性粒细胞在蓖麻毒素肺中毒小鼠炎症发展过程中对下游炎症因子的影响。方法首先基于C57小鼠采用抗Gr-1抗体构建耗竭中性粒细胞的小鼠模型,以2×LD_(50)蓖麻毒素液体气溶胶以肺递送方式对小鼠攻毒后,采用流式细胞术和肺脏组织免疫组化验证中性粒细胞耗竭效果。评价肺脏组织病理变化以及肺泡灌洗液中促炎因子和趋化因子浓度变化,从而分析中性粒细胞发挥的作用。结果流式细胞术及肺脏组织免疫组化结果显示,耗竭中性粒细胞的小鼠模型构建成功。蓖麻毒素攻毒后,与对照组小鼠相比,耗竭中性粒细胞小鼠肺脏未显示明显组织病理变化,血管周围肺水肿程度无明显差异,但肺泡灌洗液中IL-1β、IL-13、IL-17A、IL-22、IL-23、IL-1α和ENA-78促炎因子表达上调,GRO-α、MCP-1、G-CSF和M-CSF趋化因子表达上调,抑炎因子IL-10表达上调,趋化因子MIP-1β表达下调。结论与对照组小鼠相比,耗竭中性粒细胞小鼠多种促炎和趋化因子代偿性升高,提示中性粒细胞在蓖麻毒素气溶胶肺中毒后的炎症平衡中具有重要作用,可能抑制了炎症的过度发展。Objective To explore the effects of neutrophils on downstream inflammatory factors during the inflammatory development of lung ricin poisoning mice.Methods A mouse model of depleted neutrophils was first constructed based on C57 mice using anti-GR-1 antibody.After the mice were challenged by 2×LD_(50) ricin liquid aerosol via lung delivery,the effect of neutrophils depletion was verified by flow cytometry and pulmonary tissue immunohistochemical method.The neutrophilic granulocyte depletion mouse model was successfully constructed to evaluate the pathological changes of lung tissue and the changes in the concentrations of pro-inflammatory factors and chemokines in the alveolar lavage fluid,so as to analyze the role of neutrophils.Results Flow cytometry and lung tissue immunohistochemistry showed that the mouse model of depleted neutrophils was successfully constructed in this study.Poison ricin attack after depletion neutrophils mice compared with control group mice lung did not show obvious histopathological changes,no difference between the degree of pulmonary edema around blood vessels,but depletion neutrophils mice alveolar lavage in beta,IL-1β,IL-13,IL-17 A,IL-22,IL-23,IL-1αand ENA-78 proinflammatory factor expression,GRO-alpha,MCP-1,G-CSF and M-CSF chemokine expression increase,IL-10 expression suppression of inflammation factor,chemokines MIP-1 beta expression.Conclusion Compared with the control group,the depleted neutrophil mice showed increased compensative effects of various pro-inflammatory and chemokine factors,suggesting that neutrophils play an important role in the inflammatory balance after ricin aerosol pulmonary poisoning,which may inhibit the overdevelopment of inflammation.

关 键 词：蓖麻毒素 中性粒细胞 气溶胶 

分 类 号：R392.1[医药卫生—免疫学]