Clinical Application of NETs in Diagnosis and Monitoring of Acute Ischemic Stroke
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摘要:
目的 探究中性粒细胞胞外诱捕网（neutrophil extracellular traps，NETs）在急性缺血性脑卒中（acute ischemic stroke，AIS）诊断和病情监测中的价值。 方法 选取2023年6月至2024年2月昆明医科大学附属甘美医院首次确诊为AIS的患者63例作为实验组，选取同期在性别、年龄等相匹配的非AIS人群58例作为对照组，对实验组患者未经治疗前和治疗7～10 d时的NETs含量进行检测，将一般临床资料和相关实验室结果进行统计学分析。 结果 实验组的NEUT、LYMPH、MONO、EO、FDP、D-dimer以及NETs含量均显著高于对照组（P < 0.05）。患者治疗前NETs含量显著高于治疗7～10 d及对照组（P < 0.05），患者治疗7～10 d NETs含量显著高于对照组（P < 0.05）。NEUT、MONO、FDP、D-dimer、NIHSS评分与患者治疗前NETs含量正相关（P < 0.05）。患者血清NETs水平升高与AIS发病危险因素相关（P < 0.05）。NEUT、D-dimer与NETs三者联合对AIS的诊断价值优于其余指标。 结论 NETs在急性缺血性脑卒中患者血清中高表达，血清NETs对急性缺血性脑卒中的辅助诊断及病情监测具有一定价值。 -
关键词:
- 中性粒细胞胞外诱捕网 /
- 瓜氨酸组蛋白H3 /
- 中性粒细胞脂酶 /
- 髓过氧化物酶
Abstract:Objective To explore the value of neutrophil extracellular trapping nets （neutrophil extracellular traps, NETs） in the diagnosis and monitoring of acute ischemic stroke （acute ischemic stroke, AIS）. Methods From June 2023 to February 2024, 63 patients newly diagnosed with AIS at the Ganmei Hospital Affiliated with Kunming Medical University were selected as the experimental group, and 58 non-AIS individuals matched in gender, age, and other characteristics were selected as the control group. The NETs levels of patients in the experimental group were detected before and 7-10 days after treatment, and general clinical data and related laboratory results were statistically analyzed. Results The experimental group showed significantly higher levels of NEUT, LYMPH, MONO, EO, FDP, D-dimer, and NETs compared to the control group （P < 0.05）. The NETs levels before treatment were significantly higher than those at 7～10 days after treatment and in the control group （P < 0.05）. The NETs levels 7-10 days after treatment were significantly higher than those in the control group （P < 0.05）. NEUT, MONO, FDP, D-dimer, and NIHSS scores were positively correlated with the pre-treatment NETs levels （P < 0.05）. Elevated serum NETs levels were associated with AIS risk factors （P < 0.05）. The combined diagnostic value of NEUT, D-dimer, and NETs for AIS was superior to other indicators. Conclusion NETs are highly expressed in the serum of patients with acute ischemic stroke, and serum NETs have certain value in the auxiliary diagnosis and monitoring of acute ischemic stroke. -
图 4 MPO、NE、CitH3在不同组别的含量比较
n.s.统计学无差异；***P < 0.001。
Figure 4. Comparison of the levels of MPO，NE，and CitH3 in different groups
图 5 NEUT、D-D、MPO、NE、CitH3对否诊断AIS的ROC曲线
Figure 5. ROC curves of the diagnostic value of NEUT，D-D，MPO，NE，and CitH3 in AIS
图 6 MPO、NE、CitH3分别与NEUT、D-D联合检测对是否诊断AIS的ROC曲线
Figure 6. ROC curves of MPO，NE，CitH3 in combination with NEUT and D-D for the diagnosis of AIS
表 1 实验组与对照组一般临床资料及实验室指标比较[n（%）/M（P25，P75）/（$ \bar x \pm s $）]
Table 1. Comparison of general clinical data and laboratory indicators between the experimental group and the control group[n（%）/M（P25，P75）/（$ \bar x \pm s $）]
项目 实验组1（n=63） 对照组（n=58） χ2/t/Z P 性别，男 39（61.9%） 37（63.8%） 0.046 0.830 年龄（岁） 67.81 ± 13.56 63.22 ± 14.88 −1.773 0.079 NEUT 68.4（62.4，79.4） 59.6（56.5，62.2） −5.331 <0.001* LYMPH 22.1 ± 10.7 33.0 ± 5.3 7.221 <0.001* MONO 6.4（5.2，8.1） 4.6（3.6，5.9） −4.645 <0.001* EO 0.9（0.3，2.2） 1.8（1.2，2.4） −3.289 <0.001* PLT（×109/L） 214.4 ± 51.5 230.8 ± 43.3 1.894 0.061 APTT（s） 35.1（31.4，37.5） 35.9（35.2，37.2） −1.684 0.092 PT（s） 13.1（12.5，13.7） 12.9（12.5，13.4） −1.299 0.194 FIB（g/L） 3.3（2.7，3.8） 3.2（2.9，3.5） −0.496 0.620 FDP（mg/L） 2.8（1.8，5.4） 1.4（1.1，1.7） −6.688 <0.001* D-D（μg/L） 560（320， 1390 ）285（230，312.5） −6.378 <0.001* *P < 0.05。 
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表 2 实验组和对照组CitH3水平
Table 2. Citrullinated Histone H3 （CitH3） levels in the experimental and control groups
比较组别 检验统计 标准误差 标准检验统计 Sig Adj. Sig a 秩和检验 Sample 1-Sample 2 H P 实验组1-对照组 −81.969 9.692 −8.457 <0.001* <0.001* 71.353 <0.001* 实验组2-对照组 −69.048 9.692 −7.124 <0.001* <0.001* 实验组1-实验组2 12.921 9.490 1.362 0.173 0.520 *P < 0.05；a. 已针对多项检验通过 Bonferroni 校正法调整显著性值。
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表 3 实验组和对照组NE水平
Table 3. Neutrophil elastase （NE） levels in the experimental and control groups
比较组别 检验统计 标准误差 标准检验统计 Sig Adj. Sig a 秩和检验 Sample 1-Sample 2 H P 实验组1-对照组 −79.309 9.692 −8.183 <0.001* <0.001* 79.162 <0.001* 实验组2-对照组 −70.198 9.692 −7.243 <0.001* <0.001* 实验组1-实验组2 9.111 9.490 0.960 0.337 0.996 *P < 0.05；a. 已针对多项检验通过 Bonferroni 校正法调整显著性值。
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表 4 实验组和对照组MPO水平
Table 4. Myeloperoxidase （MPO） levels in the experimental and control groups
比较组别 检验统计 标准误差 标准检验统计 Sig Adj. Sig a 秩和检验 Sample 1-Sample 2 H P 实验组1-对照组 −78.597 9.692 −8.109 <0.001 <0.001 81.684 <0.001* 实验组2-对照组 −60.486 9.692 −6.241 <0.001 <0.001 实验组1-实验组2 18.111 9.490 1.909 0.056 0.169 *P < 0.05；a. 已针对多项检验通过 Bonferroni 校正法调整显著性值。
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表 5 未经治疗前血清NETs与常规实验室指标的相关性分析
Table 5. Correlation analysis of serum NETs and routine laboratory indices before treatment
组合 未治疗前MPO 未治疗前NE 未治疗前CitH3 r P r P r P NEUT 0.366 <0.001* 0.348 <0.001* 0.362 <0.001* LYMPH −0.482 <0.001* −0.487 <0.001* −0.494 <0.001* MONO 0.390 <0.001* 0.520 <0.001* 0.486 <0.001* EO −0.248 0.006* −0.215 0.018* −0.248 0.006* FDP 0.479 <0.001* 0.553 <0.001* 0.541 <0.001* D-dimer 0.350 <0.001* 0.433 <0.001* 0.431 <0.001* NIHSS评分 0.715 <0.001* 0.598 <0.001* 0.679 <0.001* *P < 0.05。
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表 6 影响AIS的多因素Logistic回归分析
Table 6. Multivariate Logistic regression analysis of factors influencing AIS
项目 赋值设置 B SE Wald P OR 95%CI 下限 上限 常量 −47.573 178.46 0.071 0.790 0.000 NEUT 实测值 0.106 0.050 4.500 0.034* 1.112 1.008 1.227 LYM 实测值 0.349 1.787 0.038 0.845 1.418 0.043 47.095 MONO 实测值 0.509 1.805 0.079 0.778 1.663 0.048 57.204 EO 实测值 0.197 2.005 0.010 0.922 1.218 0.024 62.027 FDP 实测值 0.631 0.629 1.006 0.316 1.880 0.548 6.453 Ddimer 实测值 6.634 2.871 5.338 0.021* 760.456 2.735 211418.325 NETs a 校正生成综合因子输入 4.061 1.448 7.861 0.005* 58.031 3.394 992.163 *P < 0.05；a：为解决NETs组成指标（MPO、NE、CitH3）共线问题（r > 0.7），采用三者形成的NETs综合子因子。
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表 7 NEUT、D-D、MPO、NE、CitH3对否诊断AIS的ROC曲线分析结果
Table 7. ROC curve analysis of the diagnostic value of NEUT，D-D，MPO，NE，CitH3 in the diagnosis of AIS
项目 AUC 灵敏度 特异度 约登指数 95%置信区间 P NEUT 0.781 0.746 0.81 0.556 （0.695，0.868） <0.001* D-dimer 0.836 0.746 0.879 0.625 （0.761，0.911） <0.001* 治疗前MPO 0.924 0.778 0.948 0.726 （0.880，0.969） <0.001* 治疗前NE 0.918 0.984 0.724 0.708 （0.872，0.965） <0.001* 治疗前CitH3 0.927 0.984 0.724 0.708 （0.884，0.970） <0.001* *P < 0.05。
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表 8 MPO、NE、CitH3分别与NEUT、D-D联合检测对是否诊断AIS的ROC曲线分析结果
Table 8. ROC curve analysis of the diagnostic value of MPO，NE，CitH3 in combination with NEUT and D-D for the diagnosis of AIS
项目 AUC 灵敏度 特异度 约登指数 95%CI P NEUT、D-dimer与MPO三者联合 0.988 0.968 0.948 0.916 （0.974，1.000） <0.001* NEUT、D-dimer与NE三者联合 0.970 0.937 0.966 0.903 （0.937，1.000） <0.001* NEUT、D-dimer与CitH3三者联合 0.978 0.905 0.966 0.871 （0.957，1.000） <0.001* *P < 0.05。
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