Regulating Neutrophil PAD4/NOX-Dependent Cerebrovasular Thromboinflammation.
Regulating Neutrophil PAD4/NOX-Dependent Cerebrovasular Thromboinflammation.
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DOI：
10.7150/ijbs.77434
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发表时间：
2023
影响因子：
9.2
通讯作者：
Gavins FNE
中科院分区：
文献类型：
--
作者：
Ansari J;Vital SA;Yadav S;Gavins FNE
背景资料：中性粒细胞胞外陷阱（NET）的产生与血栓炎性疾病（如镰状细胞病（SCD））的发病机制有关，导致缺血性卒中的风险增加。NET由酶肽基精氨酸脱亚胺酶4（PAD 4）和中性粒细胞衍生的活性氧物质（ROS）催化，特别是NADPH氧化酶（NOX），其与PAD 4相互作用，因此对中性粒细胞功能至关重要。然而，NOX依赖性ROS和NET在与血栓炎性疾病（如SCD）相关的加速脑微血管血栓形成中的作用尚未完全阐明，这是本研究的目的。研究方法：靶向PAD 4和NOX的体外作用使用生理学相关的NET测定用从健康志愿者（对照）和SCD患者分离的中性粒细胞进行检查。此外，在C57 BL/6和镰状转基因小鼠（STM）的脑微循环中靶向PAD 4和NOX的体内血管内效应使用光活化血栓形成模型（光/染料）结合实时荧光活体显微镜进行了评估。结果：我们发现靶向人中性粒细胞中的PAD 4和NOX显著抑制离子霉素依赖性H3cit+中性粒细胞。体内靶向PAD 4和NOX导致脑血管小动脉和小静脉中的血流停止延长。此外，我们能够通过增加STM中脑微血管中的血流停止时间，在加速血栓炎症的临床模型中复制PAD 4和NOX靶向的作用。这些发现与临床环境一致，即从SCD患者分离的中性粒细胞，其具有靶向PAD 4和NOX的H3cit+中性粒细胞产生的减弱。结论：综合考虑，我们令人信服的数据表明，PAD 4和NOX在中性粒细胞驱动的血栓炎症中发挥重要作用。靶向PAD 4和NOX限制病理性H3cit+中性粒细胞，这可以进一步解释脑血栓形成的衰减。总体而言，本研究提供了预防和管理血栓炎性并发症（如缺血性卒中）的可行临床前模型。
Background: Neutrophil extracellular trap (NET) production has been implicated in the pathogenesis of thromboinflammatory conditions such as Sickle Cell Disease (SCD), contributing to heightened risk for ischemic stroke. NETs are catalyzed by the enzyme Peptidyl Arginine Deiminase 4 (PAD4) and neutrophil derived reactive oxygen species (ROS), especially NADPH oxidase (NOX) which interacts with PAD4 and is therefore critical for neutrophil function. However, the role that NOX-dependent ROS and NETs play in the accelerated cerebral microvascular thrombosis associated with thromboinflammatory conditions, such as SCD, has not been fully elucidated and is the aim of this study. Methods: The in-vitro effects of targeting PAD4 and NOX were examined using physiologically relevant NET assays with neutrophils isolated from healthy volunteers (control) and SCD patients. In addition, in-vivo intravascular effects of targeting PAD4 and NOX in the cerebral microcirculation of C57BL/6 and sickle transgenic mice (STM) were assessed using a photoactivation thrombosis model (light/dye) coupled with real-time fluorescence intravital microscopy. Results: We found that targeting PAD4 and NOX in human neutrophils significantly inhibited ionomycin dependent H3cit+ neutrophils. Targeting PAD4 and NOX in-vivo resulted in prolonged blood flow cessation in cerebrovascular arterioles as well as venules. Moreover, we were able to replicate the effects of PAD4 and NOX targeting in a clinical model of accelerated thromboinflammation by increasing blood flow cessation times in cerebral microvessels in STM. These findings concurred with the clinical setting i.e. neutrophils isolated from SCD patients, which possessed an attenuation of H3cit+ neutrophil production on targeting PAD4 and NOX. Conclusions: Taken together, our compelling data suggests that PAD4 and NOX play a significant role in neutrophil driven thromboinflammation. Targeting PAD4 and NOX limits pathological H3cit+ neutrophils, which may further explain attenuation of cerebral thrombosis. Overall, this study presents a viable pre-clinical model of prevention and management of thromboinflammatory complications such as ischemic stroke.
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影响因子：
4.6
作者：
Arcanjo A;Logullo J;Menezes CCB;de Souza Carvalho Giangiarulo TC;Dos Reis MC;de Castro GMM;da Silva Fontes Y;Todeschini AR;Freire-de-Lima L;Decoté-Ricardo D;Ferreira-Pereira A;Freire-de-Lima CG;Barroso SPC;Takiya C;Conceição-Silva F;Savino W;Morrot A
通讯作者：
Morrot A
活性氧调节血小板活化和血栓形成。
影响因子：
11.4
作者：
Qiao J;Arthur JF;Gardiner EE;Andrews RK;Zeng L;Xu K
通讯作者：
Xu K
影响因子：
4.9
作者：
Esmon, CT
通讯作者：
Esmon, CT
影响因子：
7.3
作者：
Konig MF;Andrade F
通讯作者：
Andrade F
影响因子：
4.6
作者：
Lu, Wan Jung;Li, Jiun Yi;Lin, Kuan Hung
通讯作者：
Lin, Kuan Hung