CAJ | 학술논문

目的：本研究旨在通过体内和体外实验建立在小鼠脓毒症模型中检测中性粒细胞胞外诱捕网（NETs）的方法，并初步探讨其与脓毒症肺组织损伤的相关性。方法采用盲肠结扎穿孔手术建立脓毒症小鼠模型，以假手术小鼠作为对照组。术后7 d，采用PicoGreen?染料定量检测小鼠血浆游离DNA，采用髓过氧化物酶（MPO）-DNA ELISA检测小鼠外周NETs复合物的形成，反映NETs生成量。对脓毒症小鼠的肺组织进行病理分析，反映肺组织损伤情况。采用免疫荧光染色法对肺组织原位的NETs形成情况进行鉴定。结果与对照组相比，脓毒症小鼠血浆中游离DNA含量（1650 ng/ml）显著升高（t =21.91，P =0.0021）。MPO-DNA复合物水平（4.85倍）也显著升高（t =15.40，P=0.0042）。脓毒症小鼠的肺组织可见明显损伤，免疫荧光检测显示脓毒症小鼠肺部的NETs形成增加。结论在小鼠中建立了稳定的NETs形成情况鉴定方法，且肺部NETs形成增加是脓毒症模型小鼠肺组织损伤的原因之一。
목적：본연구지재통과체내화체외실험건립재소서농독증모형중검측중성립세포포외유포망（NETs）적방법，병초보탐토기여농독증폐조직손상적상관성。방법채용맹장결찰천공수술건립농독증소서모형，이가수술소서작위대조조。술후7 d，채용PicoGreen?염료정량검측소서혈장유리DNA，채용수과양화물매（MPO）-DNA ELISA검측소서외주NETs복합물적형성，반영NETs생성량。대농독증소서적폐조직진행병리분석，반영폐조직손상정황。채용면역형광염색법대폐조직원위적NETs형성정황진행감정。결과여대조조상비，농독증소서혈장중유리DNA함량（1650 ng/ml）현저승고（t =21.91，P =0.0021）。MPO-DNA복합물수평（4.85배）야현저승고（t =15.40，P=0.0042）。농독증소서적폐조직가견명현손상，면역형광검측현시농독증소서폐부적NETs형성증가。결론재소서중건립료은정적NETs형성정황감정방법，차폐부NETs형성증가시농독증모형소서폐조직손상적원인지일。
Objective To detect the neutrophil extracellular traps (NETs) levels and to investigate the lung injury in sepsis mouse models and sham controls. Discuss the potential mechanisms of NETs on sepsis induced lung injury. Methods The cell free DNA in the serum of sepsis mouse models and sham controls were detected with PicoGreen? dye for quantitative detection. The NETs compound content was detected by MPO-DNA ELISA. The injury of lung was determined by HE staining. The NETs formation in the lung of sepsis mouse models and sham controls were detected by immunolfuorescence method. Results Compare with the sham control, the content of cell free DNA in the serum of sepsis mouse models (1 650 ng/ml) was signiifcantly increased (t=21.91,P=0.0021). The content of MPO-DNA compound (4.85 folds) was also signiifcantly increased (t=15.40,P=0.0042). The lungs of sepsis mouse models were injured. Immunolfuorescence showed that NETs in lungs of sepsis mouse models were signiifcantly increased than sham operation controls. Conclusions Established stable methods to detect the NETs formation in mouse model. Excessive formation of NETs is one cause of lung injury in the sepsis mouse models.