肿瘤抑素片段选择性抑制实验性哮喘恶化的中性粒细胞浸润
2018/11/02
摘要
背景：哮喘是一种伴有气道结构改变慢性炎症性疾病。伯吉斯和同事最近发现，与非哮喘患者相比，成年哮喘患者肺组织中的肿瘤抑制素明显减少。ECM片段如肿瘤抑素被命名为Matrikines，并独立于母体分子而起作用。COL-IV matrikines在中性粒细胞炎症（如哮喘恶化）中的作用至今尚未被研究。成人重症哮喘表型主要表现为嗜中性粒细胞炎症和哮喘急性发作频发。
目的：本研究旨在探讨一种新的活性区在肿瘤抑制素（CP17）中的作用及其在与哮喘恶化相关的中性粒细胞炎症反应中的意义。
方法：对于活性氧产生，分离的中性粒细胞用肽或载体预孵育1h并刺激（PMA）。记录发光信号（超过10秒的积分）1.5小时，根据SIMA协议进行中性粒细胞迁移。小鼠腹腔注射腹腔注射致敏的OVA/ Alumn。然后用CP17、载体（PBS）或加扰肽（SP17）在OVA暴露后（第27天和第28天，polyI:C刺激）处理小鼠。第29天处死动物，进行肺功能测定、组织学和灌洗。
结果：CP17使总活性氧产生率降低到52.44%（0.5μm，P＜0.05 vs. SP17），降低了体外定向性（vs. SP17，P＝1x10-6）和迁移速度（5μm，p= 1x10-3）。CP17在体内应用减少中性粒细胞炎症～1.8倍（P＜0.001 vs. SP17），减少黏液产生细胞数（- 29%，P＜0.05）。
结论：CP17降低了活性氧产生率、迁移速度，并选择性抑制肺间质和肺泡腔中中性粒细胞的积累。
临床相关性：CP17可作为药物开发的潜在前体以对抗绝大多数中性粒细胞炎症。
Tumstatin fragment selectively inhibits neutrophil infiltration in experimental asthma exacerbation.
Nissen G, Hollaender H, Tang FSM, Wegmann M, Lunding L, Vock C, Bachmann A, Lemmel S, Bartels R, Oliver BG, Burgess JK, Becker T, Kopp MV, Weckmann M.
Abstract
BACKGROUND: Asthma is a chronic inflammatory disease with structural changes present. Burgess and colleagues recently found tumstatin markedly reduced in adult asthmatic lung tissue compared to non-asthmatics. ECM-fragments such as tumstatin are named matrikines and act independently of the parent molecule. The role of Col IV matrikines in neutrophil inflammation (e.g. exacerbation in asthma) have not been investigated to date. Severe adult asthma phenotypes are dominated by neutrophilic inflammation and show a high frequency of severe exacerbations.
OBJECTIVE: This study sought to investigate the role of a novel active region within tumstatin (CP17) and its implication in neutrophil inflammatory responses related to asthma exacerbation.
METHODS: For reactive oxygen production, isolated neutrophils were pre-incubated with peptides or vehicle for 1h and stimulated (PMA). Luminescence signal was recorded (integration over 10 sec) for 1.5 h. Neutrophil migration was performed according to the SiMA protocol. Mice were sensitized to OVA/Alumn by intraperitoneal (i.p.) injections. Mice were then treated with CP17, vehicle (PBS) or scrambled peptide (SP17) after OVA exposure (day 27 and 28, polyI:C stimulation). All animals were sacrificed on day 29 with lung function measurement, histology and lavage.
RESULTS: CP17 decreased total ROS production rate to 52.44% (0.5μM, p<0.05 vs. SP17), reduced the in vitro directionality (vs. SP17, p=1x10-6 ) and migration speed (5μM, p =1x10-3 ). In vivo application of CP17 decreased neutrophil inflammation ~1.8-fold (p<0.001 vs. SP17) and reduced numbers of mucus producing cells (-29%, p<0.05).
CONCLUSION: CP17 reduced the ROS production rate, migrational speed and selectively inhibited neutrophil accumulation in the lung interstitium and lumen.
CLINICAL RELEVANCE: CP17 may serve as a potential precursor for drug development to combat overwhelming neutrophil inflammation. This article is protected by copyright. All rights reserved.
背景：哮喘是一种伴有气道结构改变慢性炎症性疾病。伯吉斯和同事最近发现，与非哮喘患者相比，成年哮喘患者肺组织中的肿瘤抑制素明显减少。ECM片段如肿瘤抑素被命名为Matrikines，并独立于母体分子而起作用。COL-IV matrikines在中性粒细胞炎症（如哮喘恶化）中的作用至今尚未被研究。成人重症哮喘表型主要表现为嗜中性粒细胞炎症和哮喘急性发作频发。
目的：本研究旨在探讨一种新的活性区在肿瘤抑制素（CP17）中的作用及其在与哮喘恶化相关的中性粒细胞炎症反应中的意义。
方法：对于活性氧产生，分离的中性粒细胞用肽或载体预孵育1h并刺激（PMA）。记录发光信号（超过10秒的积分）1.5小时，根据SIMA协议进行中性粒细胞迁移。小鼠腹腔注射腹腔注射致敏的OVA/ Alumn。然后用CP17、载体（PBS）或加扰肽（SP17）在OVA暴露后（第27天和第28天，polyI:C刺激）处理小鼠。第29天处死动物，进行肺功能测定、组织学和灌洗。
结果：CP17使总活性氧产生率降低到52.44%（0.5μm，P＜0.05 vs. SP17），降低了体外定向性（vs. SP17，P＝1x10-6）和迁移速度（5μm，p= 1x10-3）。CP17在体内应用减少中性粒细胞炎症～1.8倍（P＜0.001 vs. SP17），减少黏液产生细胞数（- 29%，P＜0.05）。
结论：CP17降低了活性氧产生率、迁移速度，并选择性抑制肺间质和肺泡腔中中性粒细胞的积累。
临床相关性：CP17可作为药物开发的潜在前体以对抗绝大多数中性粒细胞炎症。
(中日友好医院呼吸与危重症医学科 李红雯 摘译 林江涛 审校)
(Clin Exp Allergy. 2018 Jul 20. doi: 10.1111/cea.13236.)
(Clin Exp Allergy. 2018 Jul 20. doi: 10.1111/cea.13236.)
Tumstatin fragment selectively inhibits neutrophil infiltration in experimental asthma exacerbation.
Nissen G, Hollaender H, Tang FSM, Wegmann M, Lunding L, Vock C, Bachmann A, Lemmel S, Bartels R, Oliver BG, Burgess JK, Becker T, Kopp MV, Weckmann M.
Abstract
BACKGROUND: Asthma is a chronic inflammatory disease with structural changes present. Burgess and colleagues recently found tumstatin markedly reduced in adult asthmatic lung tissue compared to non-asthmatics. ECM-fragments such as tumstatin are named matrikines and act independently of the parent molecule. The role of Col IV matrikines in neutrophil inflammation (e.g. exacerbation in asthma) have not been investigated to date. Severe adult asthma phenotypes are dominated by neutrophilic inflammation and show a high frequency of severe exacerbations.
OBJECTIVE: This study sought to investigate the role of a novel active region within tumstatin (CP17) and its implication in neutrophil inflammatory responses related to asthma exacerbation.
METHODS: For reactive oxygen production, isolated neutrophils were pre-incubated with peptides or vehicle for 1h and stimulated (PMA). Luminescence signal was recorded (integration over 10 sec) for 1.5 h. Neutrophil migration was performed according to the SiMA protocol. Mice were sensitized to OVA/Alumn by intraperitoneal (i.p.) injections. Mice were then treated with CP17, vehicle (PBS) or scrambled peptide (SP17) after OVA exposure (day 27 and 28, polyI:C stimulation). All animals were sacrificed on day 29 with lung function measurement, histology and lavage.
RESULTS: CP17 decreased total ROS production rate to 52.44% (0.5μM, p<0.05 vs. SP17), reduced the in vitro directionality (vs. SP17, p=1x10-6 ) and migration speed (5μM, p =1x10-3 ). In vivo application of CP17 decreased neutrophil inflammation ~1.8-fold (p<0.001 vs. SP17) and reduced numbers of mucus producing cells (-29%, p<0.05).
CONCLUSION: CP17 reduced the ROS production rate, migrational speed and selectively inhibited neutrophil accumulation in the lung interstitium and lumen.
CLINICAL RELEVANCE: CP17 may serve as a potential precursor for drug development to combat overwhelming neutrophil inflammation. This article is protected by copyright. All rights reserved.
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在轻度哮喘患者中按需使用布地奈德-福莫特罗与维持使用布地奈德的研究
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过敏性疾病和哮喘生物标志物的最新进展和亮点
