Allergic asthma is a chronic airway inflammatory response to different triggers like inhaled allergens. Excessive ATP in fluids from asthmatic patients is considered an inflammatory signal and an important autocrine/paracrine modulator of airway physiology. Here we investigated the deleterious effect of increased extracellular ATP (eATP) concentration on the mucociliary clearance (MCC) effectiveness and determined the role of ATP releasing channels during airway inflammation in an ovalbumin (OVA)-sensitized mouse model. Our allergic mouse model exhibited high levels of eATP measured in the tracheal fluid with a luciferin-luciferase assay and reduced MCC velocity determined by microspheres tracking in the trachea ex vivo. Addition of ATP had a dual effect on MCC, where lower ATP concentration (µM) increased microspheres velocity, while higher concentration (mM) transiently stopped microspheres movement. Also, an augmented ethidium bromide uptake by the allergic tracheal airway epithelium suggests an increase in ATP release channel functionality during inflammatory conditions. The use of carbenoxolone, a non-specific inhibitor of connexin and pannexin1channels reduced the eATP concentration in the allergic mouse tracheal fluid and dye uptake by the airway epithelium, providing evidence that these ATP release channels are facilitating the net flux of ATP to the lumen during airway inflammation. However, only the specific inhibition of pannexin1 with ¹⁰Panx peptide significantly reduced eATP in bronchoalveolar lavage and decreased airway hyperresponsiveness in OVA-allergic mouse model. These data provide evidence that blocking eATP may be a pharmacological alternative to be explored in rescue therapy during episodes of airflow restriction in asthmatic patients.

中文翻译：

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阻断 ATP 释放通道可防止哮喘小鼠模型中的高细胞外 ATP 水平和气道高反应性

过敏性哮喘是对吸入过敏原等不同触发因素的慢性气道炎症反应。哮喘患者体液中过量的 ATP 被认为是炎症信号和气道生理学的重要自分泌/旁分泌调节剂。在这里，我们研究了细胞外 ATP (eATP) 浓度增加对粘液纤毛清除 (MCC) 有效性的有害影响，并确定了 ATP 释放通道在卵清蛋白 (OVA) 致敏小鼠模型中气道炎症期间的作用。我们的过敏小鼠模型在气管液中显示出高水平的 eATP，通过荧光素-荧光素酶测定法测定，并通过体外气管中的微球跟踪确定 MCC 速度降低。添加 ATP 对 MCC 有双重影响，其中较低的 ATP 浓度 (μM) 增加了微球速度，而较高浓度 (mM) 会暂时停止微球运动。此外，过敏性气管气道上皮对溴化乙锭的吸收增加表明在炎症条件下 ATP 释放通道功能增加。使用卡苯氧酮（一种非特异性连接蛋白和 pannexin1 通道抑制剂）降低了过敏小鼠气管液中的 eATP 浓度和气道上皮对染料的摄取，提供了证据表明这些 ATP 释放通道正在促进 ATP 到管腔的净通量气道炎症。然而，只有 pannexin1 的特异性抑制与 过敏性气管气道上皮对溴化乙锭的吸收增加表明在炎症条件下 ATP 释放通道功能增加。使用卡苯氧酮（一种非特异性连接蛋白和 pannexin1 通道抑制剂）降低了过敏小鼠气管液中的 eATP 浓度和气道上皮对染料的摄取，提供了证据表明这些 ATP 释放通道正在促进 ATP 到管腔的净通量气道炎症。然而，只有 pannexin1 的特异性抑制与 过敏性气管气道上皮对溴化乙锭的吸收增加表明在炎症条件下 ATP 释放通道功能增加。使用卡苯氧酮（一种非特异性连接蛋白和 pannexin1 通道抑制剂）降低了过敏小鼠气管液中的 eATP 浓度和气道上皮对染料的摄取，提供了证据表明这些 ATP 释放通道正在促进 ATP 到管腔的净通量气道炎症。然而，只有 pannexin1 的特异性抑制与 提供证据表明这些 ATP 释放通道在气道炎症期间促进了 ATP 到管腔的净通量。然而，只有 pannexin1 的特异性抑制与 提供证据表明这些 ATP 释放通道在气道炎症期间促进了 ATP 到管腔的净通量。然而，只有 pannexin1 的特异性抑制与¹⁰ Panx 肽显着降低支气管肺泡灌洗液中的 eATP，并降低 OVA 过敏小鼠模型中的气道高反应性。这些数据提供的证据表明，在哮喘患者出现气流受限的情况下，阻断 eATP 可能是在抢救治疗中探索的一种药理学替代方法。