Research on asthma children′s neutrophil to form extracellular traps induced by nitric oxide donor

LI Wen-xuan, ZHU Yun-qian, WANG Xiao-ming

Chinese Journal of Child Health Care ›› 2018, Vol. 26 ›› Issue (8) : 831-834.

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Chinese Journal of Child Health Care ›› 2018, Vol. 26 ›› Issue (8) : 831-834. DOI: 10.11852/zgetbjzz2018-26-08-06

Research on asthma children′s neutrophil to form extracellular traps induced by nitric oxide donor

  • LI Wen-xuan, ZHU Yun-qian, WANG Xiao-ming
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Abstract

Objective To investigate the inductive effect of nitric oxide donor on asthmatic children′s neutrophil extracellular traps (NETs),in order to find new monitoring indicators and therapeutic targets that may be used in childhood asthma. Methods Totally 38 children with bronchial asthma in the Pediatric Department of the Fifth People′s Hospital of Shanghai were enrolled in this study from September to November in 2017. Percoll density gradient centrifugation was used to isolate neutrophils from the peripheral blood of asthmatic children, and nitric oxide donor (SNP) was added for in vitro incubation.Fluorescent staining was used to detect the double-stranded DNA (the main component of NETs) in the cell supernatant for quantitative analysis.Meantime, the cell slides were prepared to observe the main structures of NETs by fluorescence microscopy.In addition, inhibitors of nitric oxide synthase (NOS) were used to interfere the induction of SNP. Results The neutrophils were successfully isolated from the peripheral blood of asthmatic children, and the recovery rate was over 95%, the cell viability was over 80%.The extracellular mesh-like NETs structures were obviously observed in 3 hours after induction in PMA and SNP group, but NETs structure was not observed in PBS, SNP + L-NAME and SNP + SMT groups.The relative amounts of NETs in the intervention group (PMA 160 nmol/L, LPS 100 ng/ml and SNP 400 μmol/L) were significantly higher than those in the negative control PBS group (P<0.05).The relative amounts of NETs in SNP + NOS blockers group (SMT 10 mmol/L, L-NAME 500 mol/L and L-NMMA 1 mmol/L) were less than simple SNP group (P<0.05).NOS inhibitor could reduce but could not completely block the generation of NETs (P<0.05).The relative generation of NETs at 1 h, 2 h, 3 h induced by SNP kept increasing while the formation decreased at 3 h with PBS(P<0.05).With the prolongation of L-NAME, the relative amount of NETs induced by SNP decreased continuously(P<0.05). Conclusion SNP can induce the formation of NETs in asthmatic children, and inhibitors of NOS can reduce the generation of NETs, which provides a new idea for the monitoring and treatment of childhood asthma.

Key words

asthma / neutrophil extracellular traps / nitric oxide donor / nitric oxide synthase

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LI Wen-xuan, ZHU Yun-qian, WANG Xiao-ming. Research on asthma children′s neutrophil to form extracellular traps induced by nitric oxide donor[J]. Chinese Journal of Child Health Care. 2018, 26(8): 831-834 https://doi.org/10.11852/zgetbjzz2018-26-08-06

References

[1] 全国儿科哮喘协作组, 中国疾病预防控制中心环境与健康相关产品安全所.第三次中国城市儿童哮喘流行病学调查[J].中华儿科杂志, 2013,51(10):729-735.
[2] Brinkmann V, Reichard U, Goosmann C, et al.Neutrophil extracellular traps kill bacteria[J].Science, 2004,303(5663):1532-1535.
[3] Jorch SK, Kubes P.An emerging role for neutrophil extracellular traps in noninfectious disease[J].Nat Med, 2017,23(3):279-287.
[4] Akk A, Springer LE, Pham CT.neutrophil extracellular traps enhance early inflammatory response in sendai virus-induced asthma phenotype[J].Front Immunol, 2016,7:325.
[5] Dicker AJ, Crichton ML, Pumphrey EG, et al.Neutrophil extracellular traps are associated with disease severity and microbiota diversity in patients with chronic obstructive pulmonary disease[J].J Allergy Clin Immunol, 2018.141(1):117-127.
[6] Toussaint M, Jackson DJ, Swieboda D, et al.Host DNA released by NETosis promotes rhinovirus-induced type-2 allergic asthma exacerbation[J].Nat Med, 2017,23(6):681-691.
[7] Patel S, Kumar S, Jyoti A, et al. Nitric oxide donors release extracellular traps from human neutrophils by ougmention free radical generation[J]. Nitric Oxide, 2010,22(3):226-234.
[8] 中华医学会儿科学分会呼吸学组, 编辑委员会中华儿科杂志.儿童支气管哮喘诊断与防治指南(2016年版)[J].中华儿科杂志, 2016,54(3):167-181.
[9] Cortjens B, van Woensel JB, Bem RA.Neutrophil extracellular traps in respiratory disease:guided anti-microbial traps or toxic webs[J].Paediatr Respir Rev, 2017,21:54-61.
[10] Barnado A, Crofford LJ, Oates JC.At the bedside:neutrophil extracellular traps (NETs) as targets for biomarkers and therapies in autoimmune diseases[J].J Leukoc Biol, 2016,99(2):265-278.
[11] 倪睿, 魏丰贤, 丁界先, 等.中性粒细胞胞外诱捕网(NETs)的研究现状[J].免疫学杂志, 2016,32(4):356-359.
[12] Ricciardolo FLM, Sorbello V, Ciprandi G.A pathophysiological approach for FeNO:A biomarker for asthma[J].Allergol et Immunopathol, 2015,43(6):609-616.
[13] Dubey M, Nagarkoti S, Awasthi D, et al.Nitric oxide-mediated apoptosis of neutrophils through caspase-8 and caspase-3-dependent mechanism[J].Cell Death Dis, 2016,7(9):e2348.
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