NETs在急性呼吸窘迫综合征新生儿中的变化及意义

Changes and Significance of NETs in Neonates with Acute Respiratory Distress Syndrome

目的观察急性呼吸窘迫综合征(acute respiratory distress syndrome,ARDS)新生儿中是否存在中性粒细胞胞外诱捕网(neutrophil extracellular traps,NETs)的形成,以及动态观察血浆游离DNA(cell-free DNA,cf-DNA)/NETs的变化在ARDS严重程度及治疗效果中的意义。方法前瞻性选取2020年10月~2022年4月在江苏大学附属医院收治的需呼吸机辅助通气的37例ARDS新生儿为ARDS组,根据氧合指数(oxygenation index,OI)分为轻度ARDS组(4≤OI<8)、中度ARDS组(8≤OI<16)及重度ARDS组(OI≥16);以同期分娩的27例正常新生儿为对照组。采用荧光酶标仪检测cf-DNA的含量;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)分别检测NETs成分中性粒细胞弹性蛋白酶(neutrophil elastase,NE)的含量及肿瘤坏死因子(tumor necrosis factor-α,TNF-α)水平。结果ARDS各组、对照组cf-DNA、NE及TNF-α水平比较,差异均有统计学意义(F=57.517,P<0.001;F=219.408,P<0.001;F=133.200,P=0.005),ARDS各组cf-DNA、NE及TNF-α水平均明显高于对照组(P均<0.05),ARDS各组中中度组NE水平高于轻度组(P<0.05),重度组cf-DNA、NE及TNF-α水平均高于轻度组及中度组(P均<0.05)。各ARDS组治疗3、7天时的cf-DNA、NE及TNF-α水平较治疗前均有不同程度下降,差异均有统计学意义(时间:F=52.750,P<0.001;F=44.859,P<0.001;F=62.098,P<0.001;组间:F=21.237,P<0.001;F=75.427,P<0.001;F=31.776,P=0.005;交互:F=3.042,P=0.023;F=3.047,P=0.023;F=8.618,P<0.001)。ARDS患者cf-DNA与NE及TNF-α水平呈正相关(r=0.804,P<0.001;r=0.790,P<0.001)。结论新生儿ARDS存在NETs的形成,cf-DNA/NETs水平在监测新生儿ARDS病情严重程度及治疗效果中有一定的临床价值。

Objective To observe the formation of neutrophil extracellular traps(NETs)in neonates with acute respiratory distress syndrome(ARDS),and todynamically observe the significance of plasma cell-free DNA(cf-DNA)/NETs changes in the severity and therapeutic effect of ARDS.Methods A total of 37 neonates with ARDS who were admitted to Affiliated Hospital of Jiangsu University from October 2020 to April 2022 were prospectively divided into mild ARDS group(4≤OI<8),moderate ARDS group(8≤OI<16)and severe ARDS group(OI≥16)according to oxygenation index(OI).A total of 27 normal newborn who delivered at the same time was used as the control group.The content of cf-DNA was detected by fluorescence spectrophotometer.The content of neutrophil elastase(NE)and the levels of tumor necrosis factor-α(TNF-α)in NETs were detected by enzyme linked immunosorbent assay(ELISA).Results The levels of cf-DNA,NE and TNF-α in ARDS group and control group were significantly different(F=57.517,P<0.001;F=219.408,P<0.001;F=133.200,P=0.005),the levels of cf-DNA,NE and TNF-α in ARDS groups were significantly higher than those in control group(P<0.05),and NE level in moderate ARDS group was higher than that in mild ARDS group(P<0.05).The levels of cf-DNA,NE and TNF-α in severe group were higher than those in mild group and moderate group(P<0.05).The levels of cf-DNA,NE and TNF-α in ARDS groups decreased at 3d and 7d after treatment,and the differences were statistically significant(time:F=52.750,P<0.001;F=44.859,P<0.001;F=62.098,P<0.001;between groups:F=21.237,P<0.001;F=75.427,P<0.001;F=31.776,P=0.005;interaction:F=3.042,P=0.023;F=3.047,P=0.023;F=8.618,P<0.001).cf-DNA was positively correlated with NE and TNF-αlevels in ARDS patients(r=0.804,P<0.001;r=0.790,P<0.001).Conclusion NETs are formed in neonatal ARDS,and the level of cf-DNA/NETs has certain clinical value in monitoring the severity and therapeutic effect of neonatal ARDS.