Effects of β2-adrenergic agonists on periostin-induced adhesion, superoxide anion generation, and degranulation of human eosinophils
Effects of β2-adrenergic agonists on periostin-induced adhesion, superoxide anion generation, and degranulation of human eosinophils
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β2-肾上腺素能激动剂对骨膜素诱导的粘附、超氧阴离子生成和人嗜酸性粒细胞脱颗粒的影响
DOI：
10.1016/j.alit.2018.04.007
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发表时间：
2018
影响因子：
6.8
通讯作者：
Nagata M
中科院分区：
文献类型：
--
作者：
Noguchi T;Nakagome K;Kobayashi T;Ueda Y;Soma T;Nakamoto H;Nagata M
Periostin是一种细胞外基质蛋白，在哮喘患者的气道中高度表达，以响应T辅助细胞（Th）2细胞因子，包括白细胞介素（IL）-13。1骨膜蛋白已被强调为哮喘中Th 2免疫应答的生物标志物。1 e3例如，Jia等人将血清骨膜蛋白确定为重度哮喘患者气道嗜酸性粒细胞增多的单一最佳预测因子。2骨膜蛋白也作为一种基质细胞蛋白发挥作用，1即，它与细胞受体结合并激活细胞，包括嗜酸性粒细胞。我们最近报道，骨膜蛋白诱导嗜酸性粒细胞粘附，超氧阴离子（O2-）的产生，并通过aMb 2整联蛋白脱粒。4因此，骨膜蛋白和嗜酸性粒细胞之间的相互作用可能参与了Th 2介导的嗜酸性粒细胞主导型哮喘的气道炎症的发展。β-2-肾上腺素能激动剂，包括福莫特罗和沙丁胺醇，用于治疗支气管哮喘，包括Th 2介导的哮喘。我们以前报道过福莫特罗抑制嗜酸性粒细胞与细胞间粘附分子1的粘附以及由IL-5、白三烯D4或10 kDa干扰素-g诱导蛋白增强的嗜酸性粒细胞功能，这些都是通过aMb 2整联蛋白介导的。5因此，在本研究中，我们通过测量嗜酸性粒细胞增殖、O2-生成和嗜酸性粒细胞源性神经毒素（EDN）释放来检查福莫特罗或沙丁胺醇是否可以改变骨膜素诱导的嗜酸性粒细胞活化。嗜酸性粒细胞从非特应性健康供体的外周血中分离，并通过使用免疫磁珠的阴性选择，如前所述。4、5研究程序获得了琦玉医科大学附属医院伦理委员会的批准，并在样本采集前获得了捐赠者的知情同意。嗜酸性粒细胞用福莫特罗或沙丁胺醇预孵育（100 ml培养基，105个细胞/ml（100或1000 nM）孵育15分钟，然后在涂有骨膜蛋白（10 mg/ml）的塑料板中孵育20分钟。基于粘附的嗜酸性粒细胞的残余嗜酸性粒细胞过氧化物酶（EPO）活性评估嗜酸性粒细胞与骨膜蛋白的粘附。4，5如前所述，基于细胞色素C的超氧化物歧化酶（SOD）可还原性还原，检查嗜酸性粒细胞O2 β的产生。4、5在550 nm处测量威尔斯孔中细胞悬浮液的吸光度，然后在接下来的240 min内重复测量。
Periostin, an extracellular matrix protein, is highly expressed in the airways of asthmatics in response to T helper (Th) 2 cytokines, including interleukin (IL)-13. 1 Periostin has been highlighted as a biomarker of Th2 immune responses in asthma. 1 e3 For example, Jia et al. identified serum periostin as the single best predictor of airway eosinophilia in patients with severe asthma. 2 Periostin also functions as a matricellular protein, 1 ie, it binds to cellular receptors and activates cells, including eosinophils. We recently reported that periostin induces eosinophil adhesion, superoxide anion (O2 Ā) generation, and degranulation through the aMb2 integrin. 4 Therefore, the interaction between periostin and eosinophils is likely involved in the development of airway inflammation in Th2-mediated eosinophil-dominant asthma. Beta-2-adrenergic agonists, including formoterol and salbutamol, are used to treat bronchial asthma, including Th2-mediated asthma. We previously reported that formoterol suppressed eosinophil adhesion to intercellular adhesion molecule 1 as well as eosinophil functions that are enhanced by IL-5, leukotriene D4, or interferon-g-inducible protein of 10 kDa, which are all mediated through the aMb2 integrin. 5 Thus, in this study, we examined whether formoterol or salbutamol could modify periostin-induced eosinophil activation by measuring eosinophil adhesiveness, O2 Ā generation, and eosinophil-derived neurotoxin (EDN) release. Eosinophils were isolated from the peripheral blood of nonatopic healthy donors, and by negative selection using immunomagnetic beads, as described previously. 4, 5 The study procedures were approved by the Ethics Committee of Saitama Medical University Hospital, and informed consent was obtained from the donors before sample collection.Eosinophils (100 ml of medium with 105 cells/ml) were preincubated with formoterol or salbutamol (100 or 1000 nM) for 15 min, then incubated in plastic plates coated with periostin (10 mg/ml) for 20 min. Eosinophil adhesion to periostin was assessed based on the residual eosinophil peroxidase (EPO) activity of the adherent eosinophils. 4, 5 The generation of eosinophil O2 Ā was examined based on the superoxide dismutase (SOD)-inhibitable reduction of cytochrome C, as described previously. 4, 5 The absorbance of the cell suspensions in the wells was measured at 550 nm, followed by repeated measurements over the next 240 min. The maximum value during the incubation period was