中性粒细胞胞外诱捕网标志物游离DNA水平与脓毒症血小板活化的相关性

目的研究中性粒细胞胞外诱捕网（NETs）标志物游离DNA（cfDNA）水平与血小板（PLT）活化之间的相关性。 方法根据纳入标准、排除标准将2018年9至12月中国医科大学附属第一医院ICU收治的51例患者分为脓毒症组（29例）和对照组（22例）。用酶联免疫吸附测定（ELISA）方法检测患者外周血浆的cfDNA、血小板因子4（PF4）、P-选择素浓度。采用SPSS 25.0进行统计学分析,比较2组患者入ICU 24 h内生化指标、急性生理学与慢性健康状况（APACHEⅡ）评分、序贯器官衰竭评估（SOFA）评分、国际血栓与止血委员会（ISTH）评分、日本危重病协会（JAAM）评分、cfDNA以及PLT标志物（PF4、P-选择素）的差异性。同时将脓毒症组cfDNA和PLT、PLT标志物（PF4、P-选择素）、炎症指标［中性粒细胞计数（NE）、C反应蛋白（CRP）、降钙素原（PCT）］、凝血指标［凝血酶原时间（PT）、活化部分凝血酶时间（APTT）、D-二聚体（D-D）、抗凝血酶Ⅲ活性（AT-Ⅲ）］、疾病严重程度（APACHEⅡ评分、SOFA评分、ISTH评分、JAAM评分）等分别进行相关性分析。 结果脓毒症组患者cfDNA水平明显高于对照组,差异有统计学意义［（174.499±76.596）ng/ml vs （114.354±19.319）ng/ml,P＜0.05］,但2组P-选择素、PF4水平比较,差异无统计学意义（P＞0.05）。脓毒症组中cfDNA水平与P-选择素/PLT、D-D、CRP、APACHEⅡ评分、JAAM评分之间具有相关性（r=0.3849、0.3872、0.6211、0.4542、0.4451,P＜0.05）;与NE、PLT计数、PLT标志物（PF4、P-选择素）、ISTH评分等无明显相关性（P＞0.05）。 结论脓毒症组患者cfDNA明显升高。脓毒症组中cfDNA与D-D之间具有明显正相关性,提示NETs可能与继发性纤溶过程关系更加密切。尽管脓毒症组中cfDNA与P-选择素/PLT之间有正相关性,但因为P-选择素并不是特异性的PLT活化标志物,所以NETs与脓毒症PLT活化的关系仍需进一步探究。

The correlation between cell-free DNA level as a marker of neutrophils extracellular traps (NETs) and platelet activation in sepsis: a clinical study

ObjectiveTo investigate the correlation between the levels of cell-free DNA (cfDNA), a marker of neutrophils extracellular traps (NETs), and platelet activation. MethodsAccording to inclusion criteria and exclusion criteria, 51 patients were divided into sepsis group (29 cases) and control group (22 cases). Concentrations of cfDNA, PF4 and P-selectin in peripheral plasma were detected by ELISA. SPSS 25.0 was used for statistical analysis. The biochemical indicators, cfDNA and platelet markers (PF4, P-selectin), acute physiology and chronic health evaluation II score (APACHE II score), sequential organ failure score (SOFA), International Society on Thrombosis and Haemostasis (ISTH) score for DIC within 24 hours of ICU admission were compared between the groups. The correlation analysis was performed between cfDNA and platelets count, platelet markers (PF4, P-selectin), inflammatory markers (neutrophil count, CRP, PCT), coagulation test (PT, APTT, D-dimer, ATⅢ), disease severity (APACHEⅡ, SOFA score, ISTH score, JAAM score) in the sepsis group. ResultsThe level of cfDNA was significant elevated in sepsis group (174.499±76.596 vs 114.354±19.319, P＜0.05), but no significant statistical difference was found in levels of P-selectin and PF4. There were correlation between cfDNA and the P-selectin/PLT, D-dimer, C-reactive protein, APACHE Ⅱ, and JAAM in sepsis group (P＜0.05). However, it was not significantly correlated with neutrophil count, platelet count, platelet markers (PF4, P-selectin) or ISTH score (P＞0.05). ConclusioncfDNA in the sepsis group is significantly elevated. There is a significant positive correlation between cfDNA and D-dimer in the sepsis group, which suggests that NETs may be closely related to the secondary fibrinolysis process. Although there is a positive correlation between cfDNA and P-selectin / PLT in the sepsis group, because P-selectin is not a specific platelet activation marker, the relationship between NETs and sepsis platelet activation may be insufficient.