Neutrophil extracellular traps mediate neuro-immunothrombosis

Neutrophil extracellular traps are primarily composed of DNA and histones and are released by neutrophils to promote inflammation and thrombosis when stimulated by various inflammato ry reactions.Neutrophil extracellular trap formation occurs through lytic and non-lytic pathways that can be further classified by formation mechanisms.Histones,von Willebrand factor,fibrin,and many other factors participate in the interplay between inflammation and thrombosis.Neuroimmunothrombosis summarizes the intricate interplay between inflammation and thrombosis during neural development and the pathogenesis of neurological diseases,providing cutting-edge insights into post-neurotrauma thrombotic events.The blood-brain barrier defends the brain and spinal cord against external assaults,and neutrophil extracellular trap involvement in blood-brain barrier disruption and immunothrombosis contributes substantially to secondary injuries in neurological diseases.Further research is needed to understand how neutrophil extracellular traps promote blood-brain barrier disruption and immunothrombosis,but recent studies have demonstrated that neutrophil extracellular traps play a crucial role in immunothrombosis,and identified modulators of neuro-immunothrombosis.However,these neurological diseases occur in blood vessels,and the mechanisms are unclear by which neutrophil extracellular traps penetrate the blood-brain barrier to participate in immunothrombosis in traumatic brain injury.This review discusses the role of neutrophil extracellular traps in neuro-immunothrombosis and explores potential therapeutic interventions to modulate neutrophil extracellular traps that may reduce immunothrombosis and improve traumatic brain injury outcomes.

Crossroads:Pathogenic role and therapeutic targets of neutrophil extracellular traps in rheumatoid arthritis

Rheumatoid arthritis(RA)is a prevalent autoimmune disease whose main features include chronic synovial inflammation,bone destruction,and joint degeneration.Neutrophils are often considered to be the first responders to inflammation and are a key presence in the inflammatory milieu of RA.Neutrophil extracellular traps(NETs),a meshwork of DNA-histone complexes and proteins released by activated neutrophils,are widely involved in the pathophysiology of autoimmune diseases,especially RA,in addition to playing a key role in the neutrophil innate immune response.NETs have been found to be an important source of citrullinated autoantigen antibodies and inflammatory factor release,which can activate RA synovial fibroblasts(FLS)and cause joint damage.This article reviews the role of NETs in the pathophysiology of RA,demonstrating the application of multiple molecules with various therapies,with a view to informing the discovery and development of novel biomarkers and therapeutic targets for RA.

Machine learning based on metabolomics unveils neutrophil extracellular trap-related metabolic signatures in non-small cell lung cancer patients undergoing chemoimmunotherapy

BACKGROUND Non-small cell lung cancer(NSCLC)is the primary form of lung cancer,and the combination of chemotherapy with immunotherapy offers promising treatment options for patients suffering from this disease.However,the emergence of drug resistance significantly limits the effectiveness of these therapeutic strategies.Consequently,it is imperative to devise methods for accurately detecting and evaluating the efficacy of these treatments.AIM To identify the metabolic signatures associated with neutrophil extracellular traps(NETs)and chemoimmunotherapy efficacy in NSCLC patients.METHODS In total,159 NSCLC patients undergoing first-line chemoimmunotherapy were enrolled.We first investigated the characteristics influencing clinical efficacy.Circulating levels of NETs and cytokines were measured by commercial kits.Liquid chromatography tandem mass spectrometry quantified plasma metabolites,and differential metabolites were identified.Least absolute shrinkage and selection operator,support vector machine-recursive feature elimination,and random forest algorithms were employed.By using plasma metabolic profiles and machine learning algorithms,predictive metabolic signatures were established.RESULTS First,the levels of circulating interleukin-8,neutrophil-to-lymphocyte ratio,and NETs were closely related to poor efficacy of first-line chemoimmunotherapy.Patients were classed into a low NET group or a high NET group.A total of 54 differential plasma metabolites were identified.These metabolites were primarily involved in arachidonic acid and purine metabolism.Three key metabolites were identified as crucial variables,including 8,9-epoxyeicosatrienoic acid,L-malate,and bis(monoacylglycerol)phosphate(18:1/16:0).Using metabolomic sequencing data and machine learning methods,key metabolic signatures were screened to predict NET level as well as chemoimmunotherapy efficacy.CONCLUSION The identified metabolic signatures may effectively distinguish NET levels and predict clinical benefit from chemoimmunotherapy in NSCLC patients.

Hypobaric Hypoxia Aggravates Renal Injury by Inducing the Formation of Neutrophil Extracellular Traps through the NF-κB Signaling Pathway

Objective The hypersensitivity of the kidney makes it susceptible to hypoxia injury.The involvement of neutrophil extracellular traps(NETs)in renal injury resulting from hypobaric hypoxia(HH)has not been reported.In this study,we aimed to investigate the expression of NETs in renal injury induced by HH and the possible underlying mechanism.Methods A total of 24 SD male rats were divided into three groups(n=8 each):normal control group,hypoxia group and hypoxia+pyrrolidine dithiocarbamate(PDTC)group.Rats in hypoxia group and hypoxia+PDTC group were placed in animal chambers with HH which was caused by simulating the altitude at 7000 meters(oxygen partial pressure about 6.9 kPa)for 7 days.PDTC was administered at a dose of 100 mg/kg intraperitoneally once daily for 7 days.Pathological changes of the rat renal tissues were observed under a light microscope;the levels of serum creatinine(SCr),blood urea nitrogen(BUN),cell-free DNA(cf-DNA)and reactive oxygen species(ROS)were measured;the expression levels of myeloperoxidase(MPO),citrullinated histone H3(cit-H3),B-cell lymphoma 2(Bcl-2),Bax,nuclear factor kappa B(NF-κB)p65 and phospho-NF-κB p65(p-NF-κB p65)in rat renal tissues were detected by qRT-qPCR and Western blotting;the localization of NF-κB p65 expression in rat renal tissues was observed by immunofluorescence staining and the expression changes of NETs in rat renal tissues were detected by multiplex fluorescence immunohistochemical staining.Results After hypoxia,the expression of NF-κB protein in renal tissues was significantly increased,the levels of SCr,BUN,cf-DNA and ROS in serum were significantly increased,the formation of NETs in renal tissues was significantly increased,and a large number of tubular dilatation and lymphocyte infiltration were observed in renal tissues.When PDTC was used to inhibit NF-κB activation,NETs formation in renal tissue was significantly decreased,the expression level of Bcl-2 in renal tissues was significantly increased,the expression level of Bax was significantly decreased,and renal injury was significantly alleviated.Conclusion HH induces the formation of NETs through the NF-κB signaling pathway,and it promotes apoptosis and aggravates renal injury by decreasing Bcl-2 and increasing Bax expression.

Blood neutrophil extracellular traps:a novel target for the assessment of mammary health in transition dairy cows

Background:Mammary health is important for transition dairy cows and has been well recognized to exert decisive effects on animal welfare.However,the factors influencing mammary health are still unclear.Differential somatic cell count(DSCC)could reflect the mastitis risk since it is the percentage of neutrophils plus lymphocytes in total somatic cells and could be reflective of mammary health of dairy cows.This work aimed to investigate the assessment and prognosis of the health of transition cows based on blood neutrophil extracellular traps(NETs).Results:Eighty-four transition Holstein dairy cows were selected.The serum was sampled in all the animals at week 1 pre-and postpartum,and milk was sampled at week 1 postpartum.Based on the DSCC in milk at week 1,cows with lower(7.4%±4.07%,n=15)and higher(83.3%±1.21%,n=15)DSCCs were selected.High DSCC cows had higher levels of red blood cell counts(P<0.05),hemoglobin(P=0.07),and hematocrit(P=0.05),higher concentrations of serum oxidative variables[reactive oxygen species(P<0.05),malondialdehyde(P<0.05),protein carbonyl(P<0.05),and 8-hydroxy-2-deoxyguanosine(P=0.07)],higher levels of serum and milk NETs(P<0.05)and blood-milk barrier indicators,including serumβ-casein(P=0.05)and milk immunoglobulin G2(P=0.09),than those of low DSCC cows.In addition,lower concentrations of serum nutrient metabolites(cholesterol and albumin)(P<0.05)and a lower level of serum deoxyribonuclease I(P=0.09)were observed in high DSCC cows than in low DSCC cows.Among the assessments performed using levels of the three prepartum serum parameters(NETs,deoxyribonuclease I andβ-casein),the area under the curve(0.973)of NETs was the highest.In addition,the sensitivity(1.00)and specificity(0.93)were observed for the discrimination of these cows using NETs levels with a critical value of 32.2 ng/mL(P<0.05).Conclusions:The formation of NETs in blood in transition dairy cows may damage the integrity of the blood-milk barrier and thereby increase the risk for mastitis in postpartum cows.

蝮蛇咬伤对中性粒细胞NETs和炎症损伤影响的临床研究

目的探讨蝮蛇咬伤后中性粒细胞胞外诱捕网(neutrophil extracellular traps,NETs)标志物嗜中性粒细胞弹性蛋白酶(neutrophil elastase,NE)以及细胞游离DNA(cell free DNA,cf-DNA)变化和与炎症标志物及组织损伤标志物的相关性。方法选取2021年9月—2022年12月杭州市中医院皮肤科收治的蝮蛇咬伤患者作为研究对象,研究包括36例蝮蛇咬伤患者,20例健康体检者;比较两组中性粒细胞、淋巴细胞计数、嗜中性粒细胞弹性蛋白酶(neutrophil elastase,NE)、细胞游离DNA(cell free DNA,cf-DNA)、血清淀粉样蛋白A(serum amyloid A,SAA)、C反应蛋白(C-reactive protein,CRP)、中性粒细胞淋巴细胞比值(neutrophil-to-lymphocyte ratio,NLR)、肌酸激酶(creatine kinase,CK)、肌酸肌酶同工酶(creatine kinase-MB,CKMB)、天门冬氨酸转氨酶(aspartic transaminase,AST)和乳酸脱氢酶(lactate dehydrogenase,LDH)水平,计算Pearson相关系数并利用Python软件编写代码绘制热力图、箱线图,分析中性粒细胞计数与NETs标志物、炎症标志物和组织损伤标志物之间的相关性。结果蝮蛇咬伤后中性粒细胞计数升高,NETs标志物NE、cf-DNA升高,CRP、SAA、NLR、AST升高,淋巴细胞计数降低。蝮蛇咬伤后中性粒细胞数值与NE酶、cf-DNA血清含量呈正相关,而与炎标志物CRP和SAA之间没有明显相关性;而CRP与SAA、CKMB与LDH强正相关,CK与NE、SAA与NE之间呈正相关,淋巴细胞数与NLR呈负相关。结论蝮蛇咬伤可能引起NETs水平上升,促进炎症加重和组织损伤。

利多卡因通过抑制NETs改善阿霉素引起的心脏毒性

目的通过动物实验验证利多卡因抑制中性粒细胞胞外诱捕网(NETs)减轻阿霉素引起的心脏毒性。方法将30只C57BL/6小鼠随机分为对照组、阿霉素组(DOX组)和利多卡因+阿霉素组(LIDO+DOX组),每组10只。对照组单次腹腔注射生理盐水1 mL/100 g;DOX组单次腹腔注射阿霉素15 mg/kg;LIDO+DOX组尾静脉注射利多卡因6 mg/kg,30 min后腹腔注射阿霉素15 mg/kg。于第10天,取小鼠血清检测各组小鼠磷酸肌酸激酶(CK)、磷酸肌酸激酶同工酶(CK-MB)水平;采用酶联免疫吸附试验(ELISA)检测各组小鼠血浆NETs标志物游离DNA(cf-DNA)、瓜氨酸组蛋白3(CitH3)、髓过氧化物酶-DNA(MPO-DNA)的水平;采用免疫荧光法检测各组小鼠心脏组织NETs相关标记物中CitH3、髓过氧化物酶(MPO)的表达量。结果DOX组小鼠血清CK、CK-MB水平明显高于对照组和LIDO+DOX组小鼠(P<0.05)。ELISA检测结果显示,DOX组与LIDO+DOX组小鼠血浆MPO-DNA、CitH3和cf-DNA水平明显高于对照组(P<0.05),但LIDO+DOX组小鼠血浆MPO-DNA、CitH3和cf-DNA水平明显低于DOX组(P<0.05)。免疫荧光法检测结果发现,DOX组小鼠心肌组织中MPO和CitH3表达量明显高于对照组和LIDO+DOX组(P<0.05)。结论利多卡因可能是通过抑制NETs的表达来缓解阿霉素引起的心脏毒性。

Neutrophil extracellular traps mediate the crosstalk between glioma progression and the tumor microenvironment via the HMGB1/RAGE/IL-8 axis

Objective:Neutrophil extracellular traps(NETs)produced by tumor-infiltrating neutrophils(TINs)are associated with poor prognosis in patients with several types of cancer.However,the mechanisms underlying the involvement of NETs in glioma progression remain largely unknown.This study aimed to elucidate the roles of NETs in biological processes that drive the crosstalk between glioma progression and the tumor microenvironment.Methods:Neutrophil infiltration and NETs formation were investigated in glioma tissue through immunohistochemistry,and their relationships with clinicopathological features and outcomes were statistically evaluated.The effects of NETs on glioma cell progression were studied in a co-culture system.In vivo and in vitro experiments validated the reactive oxygen species activity and cytokine production of TINs,as well as the ERK signaling pathway activation and the metastasis of gliomas.Results:Neutrophil infiltration and NETs formation were induced in high-grade glioma compared with low-grade glioma.NETs induced by TINs were determined to be an oncogenic marker of high-grade gliomas and to be involved in cell proliferation and invasion.NETs overproduction promoted glioma cell proliferation,migration,and invasion.Furthermore,HMGB1 was found to bind to RAGE and activate the NF-κB signaling pathway in vitro.In addition,NETs stimulated the NF-κB signaling pathway,thus promoting IL-8 secretion in glioblastoma.Subsequently,IL-8 recruited neutrophils which in turn mediated NETs formation via the PI3 K/AKT/ROS axis in TINs.Conclusions:Our results suggest that NETs produced by TINs mediate the crosstalk between glioma progression and the tumor microenvironment by regulating the HMGB1/RAGE/IL-8 axis.Targeting NETs formation or IL-8 secretion may be an effective approach to inhibit glioma progression.

Do neutrophil extracellular traps contribute to the heightened risk of thrombosis in inflammatory diseases?

Thrombotic events,both arterial and venous,are a major health concern worldwide. Further,autoimmune diseases,such as systemic lupus erythematosus,anti-neutrophil cytoplasmic antibody(ANCA)-associated vasculitis,and antiphospholipid syndrome,predispose to thrombosis,and thereby push the risk for these morbid events even higher. In recent years,neutrophils have been identified as important players in both arterial and venous thrombosis. Specifically,chromatin-based structures called neutrophil extracellular traps(NETs) play a key role in activating the coagulation cascade,recruiting platelets,and serving as scaffolding upon which the thrombus can be assembled. At the same time,neutrophils and NETs are emerging as important mediators of pathogenic inflammation in the aforementioned autoimmune diseases. Here,we first review the general role of NETs in thrombosis. We then posit that exaggerated NET release contributes to the prothrombotic diatheses of systemic lupus erythematosus,ANCA-associated vasculitis,and antiphospholipid syndrome.

Neutrophil extracellular traps participate in the development of cancer-associated thrombosis in patients with gastric cancer

BACKGROUND The development of venous thromboembolism(VTE) is associated with high mortality among gastric cancer(GC) patients. Neutrophil extracellular traps(NETs) have been reported to correlate with the prothrombotic state in some diseases, but are rarely reported in GC patients.AIM To investigate the effect of NETs on the development of cancer-associated thrombosis in GC patients.METHODS The levels of NETs in blood and tissue samples of patients were analyzed by ELISA, flow cytometry, and immunofluorescence staining. NET generation and hypercoagulation of platelets and endothelial cells(ECs) in vitro were observed by immunofluorescence staining. NET procoagulant activity(PCA) was determined by fibrin formation and thrombin–antithrombin complex(TAT) assays.Thrombosis in vivo was measured in a murine model induced by flow stenosis in the inferior vena cava(IVC).RESULTS NETs were likely to form in blood and tissue samples of GC patients compared with healthy individuals. In vitro studies showed that GC cells and their conditioned medium, but not gastric mucosal epithelial cells, stimulated NET release from neutrophils. In addition, NETs induced a hypercoagulable state of platelets by upregulating the expression of phosphatidylserine and Pselectin on the cells. Furthermore, NETs stimulated the adhesion of normal platelets on glass surfaces. Similarly, NETs triggered the conversion of ECs to hypercoagulable phenotypes by downregulating the expression of their intercellular tight junctions but upregulating that of tissue factor. Treatment of normal platelets or ECs with NETs augmented the level of plasma fibrin formation and the TAT complex. In the models of IVC stenosis, tumor-bearing mice showed a stronger ability to form thrombi, and NETs abundantly accumulated in the thrombi of tumorbearing mice compared with control mice. Notably, the combination of deoxyribonuclease I,activated protein C, and sivelestat markedly abolished the PCA of NETs.CONCLUSION GC-induced NETs strongly increased the risk of VTE development both in vitro and in vivo. NETs are potential therapeutic targets in the prevention and treatment of VTE in GC patients.

新生儿急性呼吸窘迫综合征中NETs与MAC-1的相关性及临床意义

目的:探讨新生儿急性呼吸窘迫综合征(ARDS)中性粒细胞胞外诱捕(NETs)与巨噬细胞1抗原(MAC-1)的相关性及临床意义。方法:选取32例ARDS患儿为ARDS组,根据病情分为轻度组(n=12)、中度组(n=11)及重度组(n=9);另选同期24例黄疸新生儿为对照组。比较ARDS组和对照组患者及ARDS中不同病情患儿血清中性粒细胞弹性蛋白酶(NE)、游离DNA(cf-DNA)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、MAC-1水平;通过Pearson相关性分析观察ARDS组患儿NE及cf-DNA与TNF-α、IL-6及MAC-1水平的相关性;ROC曲线分析血清NE和cf-DNA水平对新生儿ARDS的预测价值。结果:ARDS组患儿MAC-1表达水平增强(P<0.05);NE、cf-DNA、TNF-α、IL-6高于对照组(P<0.05);ARDS组不同病情患儿NE、cf-DNA、TNF-α、IL-6水平差异有统计学意义(P<0.05),且重度组>中度组>轻度组。相关性分析显示,ARDS组患儿血清NE及cf-DNA与TNF-α、IL-6及MAC-1均呈正相关关系(P<0.05)。ROC曲线分析显示,NE、cf-DNA预测新生儿ARDS的曲线下面积(AUC)分别为0.765、0.821,截断值分别为347.5 pg/mL、138.17 ng/mL。结论:新生儿ARDS患儿NE及cf-DNA与TNF-α、IL-6及MAC-1均呈正相关关系,临床可通过监测NE、cf-DNA水平来评估新生儿ARDS病情严重程度。

Neutrophil extracellular traps in gastrointestinal cancer

Gastrointestinal(GI)cancer is a high-risk malignancy and is characterized by high mortality and morbidity worldwide.Neutrophil extracellular traps(NETs),a weblike structure consisting of chromatin DNA with interspersed cytoplasmic and granule proteins,are extruded by activated neutrophils to entrap and kill bacteria and fungi.However,accumulating evidence shows that NETs are related to the progression and metastasis of cancer.In clinical studies,NETs infiltrate primary GI cancer tissues and are even more abundant in metastatic lesions.The quantity of NETs in peripheral blood is revealed to be associated with ascending clinical tumour stages,indicating the role of NETs as a prognostic markers in GI cancer.Moreover,several inhibitors of NETs or NET-related proteins have been discovered and used to exert anti-tumour effects in vitro or in vivo,suggesting that NETs can be regarded as targets in the treatment of GI cancer.In this review,we will focus on the role of NETs in gastric cancer and colorectal cancer,generalizing their effects on tumour-related thrombosis,invasion and metastasis.Recent reports are also listed to show the latest evidences of how NETs affect GI cancer.Additionally,notwithstanding the scarcity of systematic studies elucidating the underlying mechanisms of the interaction between NETs and cancer cells,we highlight the potential importance of NETs as biomarkers and anti-tumour therapeutic targets.

Neutrophil extracellular traps in the intestinal mucosa of Eimeria-infected animals

Objective:To investigate the presence of neutrophil extracellular traps(NETs) in vivo by analysing intestinal sections from experimentally Eimeria bovis-and naturally Eimeria arloingi-infected animals.Methods:Intestinal samples of Eimeria arloingi-and Eimeria bovis-infected animals were analysed by using immunohistochemical and fluorescence approach by using monoclonal antibodies.Results:Classical NET components were confirmed by co-localization of extracellular DNA being decorated with neutrophil elastase and histones in Eimeria-infected tissue samples.Here,extrusion of NETs was exclusively detected in intestinal polymorphonuclear neutrophils infiltrating Eimeria-infected sites.In vivo NETs were either found in close proximity or in direct contact to different Eimeria stages suggesting a stage-independent process.NETs were also found within the gut lumen driven by polymorphonuclear neutrophils that were contacting released oocysts.Conclusions:We postulate that NETs might play an important role in innate defence reactions in coccidiosis therefore significantly altering the outcome of infection.

血清NETs、FIB、miR-374a-5p水平与膝关节置换术后下肢深静脉血栓形成的相关性分析

目的探讨膝关节置换术后血清中性粒细胞胞外陷阱(NETs)、纤维蛋白原(FIB)、微小RNA-374a-5p(miR-374a-5p)水平变化,并分析其与术后下肢深静脉血栓形成(DVT)相关性及其预测效能。方法选取2020年9月至2023年5月于郑州阳城医院行膝关节置换术患者108例为研究对象,依据术后7 d是否发生下肢DVT分为DVT组32例、非DVT组76例。对比分析两组临床资料及术前、术后1 d、术后3 d血清NETs、FIB、miR-374a-5p水平。采用Pearson法分析术前血清NETs、FIB、miR-374a-5p水平与静脉血栓栓塞症风险评估量表(Caprini)评分相关性。采用多因素Logistic回归分析术后下肢DVT发生的影响因素。采用受试者工作特征曲线(ROC)及曲线下面积(AUC)评价血清NETs、FIB、miR-374a-5p水平对术后下肢DVT发生的预测价值。结果DVT组术后1 d、3 d血清NETs、FIB、miR-374a-5p水平高于非DVT组(P<0.05);术前血清NETs、FIB、miR-374a-5p水平与Caprini评分呈正相关(P<0.05);Caprini评分与术后3 d血清NETs、FIB、miR-374a-5p水平升高为术后下肢DVT发生的独立危险因素(P<0.05);术后3 d血清NETs、FIB、miR-374a-5p水平联合预测术后下肢DVT发生的AUC大于单项指标预测(P<0.05)。结论膝关节置换术后DVT者血清NETs、FIB、miR-374a-5p水平升高,联合检测其水平对术后下肢DVT发生具有一定预测价值。

老年下肢深静脉血栓患者血清Gas6、IL-1β、NETs水平与介入治疗效果的关系

目的探讨老年下肢深静脉血栓(DVT)患者血清生长停滞特异性基因产物6(Gas6)、白细胞介素-1β(IL-1β)、中性粒细胞胞外陷阱(NETs)水平与介入治疗效果的关系。方法选取2021年2月至2023年1月该院收治的120例老年DVT患者为研究对象,根据治疗7 d后临床疗效分为无效组(22例)和有效组(98例)。比较两组治疗前及治疗3、7 d后血清Gas6、IL-1β、NETs水平,分析治疗3、7 d后血清Gas6、IL-1β、NETs水平与治疗效果的关系。结果无效组与有效组Gas6、IL-1β、NETs水平存在时间与组间的交互效应,差异有统计学意义(F交互=15.214,P<0.001);两组Gas6、IL-1β、NETs水平随着治疗时间变长而逐渐降低,差异均有统计学意义(F=12.375、12.271、13.277,P<0.001);无效组Gas6、IL-1β、NETs水平明显高于有效组,差异均有统计学意义(F=23.537、22.851、23.213,P<0.001)。治疗3、7 d后血清Gas6、IL-1β、NETs联合检测预测DVT患者介入治疗无效的曲线下面积明显大于各项指标单独检测(P<0.05)。结论血清Gas6、IL-1β、NETs水平对预后有明显影响,3项指标联合检测可作为预测DVT患者介入治疗效果的重要辅助途径。

Inhibiting the formation of neutrophil extracellular trapping: a potential mechanism of Chinese medicine in the treatment of ischemic stroke

Ischemic stroke(IS)is the main killer that endangers the health and life of middle-aged and elderly people worldwide.Inflammatory response plays a key regulatory role in the pathogenesis of IS.After cerebral ischemia,leukocytes rapidly accumulate,penetrate blood vessels and infiltrate brain tissue,thereby activating pro-inflammatory factors in the infarct area to exacerbate nerve damage.Neutrophil extracellular traps(NETs)are fibrous mesh structures released by activated neutrophils outside the cell,which can clear pathogens and cell debris,induce inflammatory responses and exacerbate cerebral ischemia-reperfusion(CI/R)injury.Various traditional Chinese medicines and their main components can improve neurological function defects after IS,and inhibit the formation of NETs,which opens up a new direction for the study of traditional Chinese medicines in the prevention and treatment of IS.

中性粒细胞及NETs在骨与软组织肉瘤中的作用

中性粒细胞是肿瘤微环境的重要成分之一。中性粒细胞胞外诱捕网(NETs)是中性粒细胞释放的网状结构,其形成过程被称为中性粒细胞炎性凋亡(NETosis)。肿瘤微环境中的中性粒细胞即肿瘤相关中性粒细胞(TANs),包括N1和N2两种表型,对肿瘤具有双向调控作用。其中N1型抑制肿瘤生长,N2型促进肿瘤生长,N1型与N2型受肿瘤微环境影响会发生相互转化。中性粒细胞及NETs在骨与软组织肉瘤中也发挥重要作用,可以影响肿瘤进展并能预测患者预后,阻断NETs形成可能是潜在的治疗靶点。

IL-33、IL-8和NETs在慢性阻塞性肺疾病患者肺组织中的表达及其意义

目的:探讨白细胞介素33(IL-33)、白细胞介素8(IL-8)和中性粒细胞胞外诱捕网(NETs)的主要成分髓过氧化物酶(MPO)及瓜氨酸化组蛋白3(CitH3)在慢性阻塞性肺疾病(COPD)患者肺组织中的表达情况,阐明IL-33、IL-8和NETs对COPD发生发展的影响。方法:选取因肺部结节或肺部肿瘤行肺叶切除术的77例患者作为研究对象,分为不吸烟对照组20例、吸烟对照组18例,不吸烟COPD组19例和吸烟COPD组20例。收集研究对象的肺组织标本,HE染色观察各组患者肺组织病理形态表现,免疫组织化学染色检测各组患者肺组织中IL-33、IL-8、CitH3和MPO蛋白的表达。采用Spearman相关分析COPD组患者肺组织中IL-33、IL-8、CitH3和MPO的蛋白表达水平之间及其与肺功能指标、平均内衬间隔(MLI)、平均肺泡数(MAN)和Bosken评分的相关性。结果:吸烟对照组患者气道Bosken评分明显高于不吸烟对照组(P<0.001);不吸烟COPD组患者气道Bosken评分和MLI明显高于不吸烟对照组和吸烟对照组(P<0.05),MAN明显低于不吸烟对照组和吸烟对照组(P<0.05);吸烟COPD组患者气道Bosken评分明显高于不吸烟对照组、吸烟对照组和不吸烟COPD组(P<0.05),MLI明显高于不吸烟对照组和吸烟对照组(P<0.05),MAN明显低于不吸烟对照组和吸烟对照组(P<0.05)。免疫组织化学染色,吸烟COPD组患者肺组织中IL-33蛋白表达水平明显高于不吸烟对照组、吸烟对照组和不吸烟COPD组(P<0.05),IL-8、CitH3和MPO蛋白表达水平明显高于不吸烟对照组和吸烟对照组(P<0.05);不吸烟COPD组患者肺组织中IL-33、IL-8、CitH3和MPO蛋白表达水平明显高于不吸烟对照组和吸烟对照组(P<0.05);Spearman相关性分析,COPD组患者肺组织中IL-33蛋白表达水平与第1秒用力呼气容积占用力肺活量的百分比(FEV1/FVC)、第1秒用力呼气容积占预计值百分比(FEV1%pred)和MAN等呈负相关关系(r=-0.406,P<0.05;r=-0.493,P<0.01;r=-0.567,P<0.05),与Bosken评分和MLI等呈正相关关系(r=0.935,P<0.001;r=0.590,P<0.001);COPD组患者肺组织中IL-8蛋白表达水平与FEV1/FVC、FEV1%pred和MAN等呈负相关关系(r=-0.527,P<0.01;r=-0.497,P<0.01;r=-0.463,P<0.01),与Bosken评分和MLI等呈正相关关系(r=0.557,P<0.001;r=0.486,P<0.01);COPD组患者肺组织中CitH3蛋白表达水平与FEV1/FVC、FEV1%pred和MAN等呈负相关关系(r=-0.527,P<0.01;r=-0.640,P<0.001;r=-0.531,P<0.01),与Bosken评分和MLI等呈正相关关系(r=0.565,P<0.001;r=0.585,P<0.001);COPD组患者肺组织中MPO蛋白表达水平与FEV1/FVC呈负相关关系(r=-0.329,P<0.05),与Bosken评分呈正相关关系(r=0.410,P<0.05);COPD组患者肺组织中IL-8蛋白表达水平与CitH3和MPO蛋白表达水平呈正相关关系(r=0.390,P<0.05;r=0.349,P<0.05);COPD组患者肺组织中IL-33蛋白表达水平与IL-8、CitH3和MPO蛋白表达水平呈正相关关系(r=0.602,P<0.001;r=0.616,P<0.001;r=0.387,P<0.05)。结论:IL-33、IL-8和NETs在COPD患者肺组织中的表达水平升高,三者可能参与COPD的慢性炎症,并与疾病严重程度相关。

NETs、IL-35及凝血四项在新生儿呼吸窘迫综合征患儿中的变化及与预后的关联性

目的探究中性粒细胞胞外诱捕网(NETs)、白细胞介素(IL)-35及凝血四项在新生儿呼吸窘迫综合征(NRDS)患儿中的变化及与预后的关联性。方法回顾性纳入2021年2月至2023年2月在恩施土家族苗族自治州中心医院接受治疗的NRDS患儿87例作为研究组,并随机选取87名健康新生儿作为对照组,比较组间受试对象的NETs[游离DNA(cf-DNA)、中性粒细胞弹性蛋白酶(NE)、IL-6、肿瘤坏死因子-α(TNF-α)]、IL-35及凝血四项指标[凝血活酶时间(TT)、活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)、纤维蛋白原(FIB)]情况。并对研究组患儿进行随访,将21例住院期间发生死亡的患儿纳入死亡组,其他66例患儿纳入生存组,分析组间NETs、IL-35及凝血四项指标情况,并采用受试者操作特征(ROC)曲线分析NETs指标及IL-35水平对患儿预后死亡的预测效能。结果研究组儿童的cf-DNA、NE、IL-6、TNF-α水平分别为(144.26±33.12)ng/mL、(325.15±18.26)pg/mL、(41.12±5.16)pg/mL、(41.02±5.11)pg/mL,均高于对照组,研究组的IL-35水平为(55.79±11.06)mg/L,低于对照组,差异均有统计学意义(P<0.05)。研究组儿童的APTT、PT、FIB水平分别为(34.33±5.12)s、(18.45±4.15)s、(3.56±0.67)g/L,均高于对照组,差异均有统计学意义(P<0.05),组间TT水平比较,差异无统计学意义(P>0.05)。死亡组患儿的cf-DNA、NE、IL-6、TNF-α水平分别为(161.12±18.46)ng/mL、(344.26±23.12)pg/mL、(43.55±3.11)pg/mL、(45.12±6.12)pg/mL,均高于生存组,死亡组IL-35水平为(33.79±6.11)mg/L,低于生存组,差异均有统计学意义(P<0.05)。生存组和死亡组患儿组间TT、APTT、PT、FIB水平比较,差异均无统计学意义(P>0.05)。经ROC曲线分析,可知cf-DNA、NE、IL-6、TNF-α、IL-35均对患儿的预后死亡具有较高的预测价值,其曲线下面积(AUC)值分别为0.799、0.830、0.983、0.752、0.924,预测效能较高(P<0.05)。结论NETs、IL-35及凝血四项在NRDS患儿中具有显著变化,且NETs指标及IL-35水平对患儿预后的预测效能较高,可作为评估NRDS患儿预后的重要生物标志物。

2型糖尿病患者血浆中NETs标志物cfDNA,CitH3和MPO-DNA表达水平与视网膜病变程度相关性研究

目的探讨2型糖尿病(type 2 diabetes mellitus,T2DM)患者血浆中性粒细胞外陷阱(neutrophil extracellular traps,NETs)标志物循环游离DNA(circulating free DNA,cfDNA)、髓过氧化物酶(myeloperoxidase,MPO)-DNA、瓜氨酸化组蛋白3(citrullinated histone 3,CitH3)表达水平与视网膜病变程度的相关性。方法选取2023年1~9月河南科技大学第一附属医院80例T2DM患者为研究对象,依据T2DM患者视网膜病变情况分为无糖尿病视网膜病变(non-diabetic retinopathy,NDR)组(n=30)、非增生型糖尿病视网膜病变(nonproliferative diabetic retinopathy,NPDR)组(n=26)和增生型糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)组(n=24)。收集入选患者一般资料,检测血浆cfDNA,MPO-DNA及CitH3水平;采用Spearman相关性分析cfDNA,CitH3,MPO-DNA与T2DM患者视网膜病变严重程度的关系;ROC曲线分析cfDNA,CitH3,MPO-DNA及联合预测T2DM患者糖尿病视网膜病变(diabetic retinopathy,DR)发生的效能。结果NDR组、NPDR组和PDR组cfDNA(65.13±18.28,83.34±20.10,114.52±32.36ng/ml),CitH3(17.76±4.24,22.03±5.01,26.06±8.13 ng/ml),MPO-DNA(0.31±0.04,0.35±0.05,0.45±0.05)水平依次升高,差异具有统计学意义(F=28.755,13.335,62.470,均P<0.001);其中PDR组、NPDR组cfDNA,CitH3,MPO-DNA水平均高于NDR组(t=7.076,4.837,11.436;3.550,3.455,3.324),且PDR组cfDNA,CitH3,MPODNA水平高于NPDR组(t=4.127,2.128,7.065),差异具有统计学意义(均P<0.05)。Spearman相关性分析显示,cfDNA,CitH3,MPO-DNA与T2DM患者视网膜病变严重程度均呈正相关(r=0.659,0.470,0.744,均P<0.001)。ROC曲线显示,cfDNA,CitH3,MPO-DNA及联合检测曲线下面积[AUC(95%CI)]分别为0.846(0.758~0.934),0.776(0.674~0.878),0.847(0.764~0.930)和0.938(0.889~0.987),对于T2DM患者发生DR具有一定预测价值。结论DR患者血浆中NETs标志物cfDNA,MPO-DNA及CitH3水平升高,其水平与DR程度呈正相关,一定程度上可预测T2DM患者DR的发生情况,且联合预测价值最高。