IL-13 desensitizes β2-adrenergic receptors in human airway epithelial cells through a 15-lipoxygenase/G protein receptor kinase 2 mechanism.
IL-13 desensitizes β2-adrenergic receptors in human airway epithelial cells through a 15-lipoxygenase/G protein receptor kinase 2 mechanism.
复制标题
IL-13通过15-脂氧合酶/G蛋白受体激酶2机制在人气道上皮细胞中脱敏的β2-肾上腺素受体。
DOI：
10.1016/j.jaci.2015.02.006
复制
发表时间：
2015-05
期刊：
影响因子：
--
通讯作者：
Wenzel SE
中科院分区：
文献类型：
--
作者：
Albano GD;Zhao J;Etling EB;Park SY;Hu H;Trudeau JB;Profita M;Wenzel SE
Beta-2 肾上腺素能受体 (β2AR) 激动剂是治疗哮喘的关键药物。然而，受体脱敏可能导致治疗效果丧失。虽然重复使用 β2 激动剂的脱敏作用已得到充分研究，但 2 型炎症也可能影响 β2AR 功能。评估 2 型细胞因子 IL-13 对人气道上皮细胞 (HAEC) 中 β2AR 脱敏的影响，并确定 15-脂氧合酶-1 (15LO1) 与磷脂酰乙醇胺结合蛋白-1 (PEBP1) 的结合是否通过释放 G 蛋白受体激酶-2 (GRK2) 来促进脱敏。用 ISO（10 分钟）刺激气液界面（ALI）培养物中的 HAEC，使用/不使用 IL-13（48 小时）或异丙肾上腺素（ISO）（30 分钟）预处理。通过 ELISA 测定 cAMP，通过蛋白质印迹测定 β2AR 和 GRK2 磷酸化。 siRNA 用于 15LO1 敲低。通过免疫沉淀/蛋白质印迹和免疫荧光检测GRK2、PEBP1和15LO1的相互作用。评估了对照组和哮喘患者的 HAEC 和气道组织的 I5LO1、PEBP1 和 GRK2。与单独使用 ISO 10 分钟相比，用 ISO 或 IL-13 预处理可减少 ISO 诱导的 cAMP 生成，同时β2AR 和 GRK2 磷酸化也会增加。 ISO 10 分钟后，GRK2 与 PEBP1 相关，并与低 pGRK2 水平相关。相反，在存在IL-13+ISO（10分钟）的情况下，GRK2与PEBP1的结合减少，而15LO1和pGRK2的结合增加。 15LO1 敲低可恢复 ISO 诱导的 cAMP 生成。这些发现在新鲜的哮喘细胞和组织的 HAEC 中得到了重现。 IL-13 治疗 HAEC 会导致 β2AR 脱敏，其中涉及 15LO1/PEBP1 与游离 GRK2 的相互作用，并使其磷酸化（并脱敏）β2AR，这表明 β2 激动剂的有益作用可能会在 2 型相关哮喘中减弱。
Beta-2 adrenergic receptor (β2AR) agonists are critical treatments for asthma. However, receptor desensitization can lead to loss of therapeutic effects. While desensitization to repeated use of β2 agonists is well studied, Type-2 inflammation could also impact β2AR function. To evaluate the impact of the Type-2 cytokine, IL-13, on β2AR desensitization in human airway epithelial cells (HAECs) and determine whether 15-Lipoxygenase-1 (15LO1) binding with phosphatidylethanolamine binding protein-1 (PEBP1) contributes to desensitization through release of G-protein Receptor Kinase-2 (GRK2). HAECs in air-liquid-interface (ALI) culture with/without IL-13 (48 hrs) or isoproterenol (ISO) (30 min) pretreatment were stimulated with ISO (10min). cAMP was measured by ELISA and β2AR and GRK2 phosphorylation by Western Blot. siRNA was utilized for 15LO1 knockdown. Interactions of GRK2, PEBP1 and 15LO1 were detected by Immunoprecipitation/Western Blot and immunofluorescence. HAECs and airway tissue from controls and asthmatics were evaluated for I5LO1, PEBP1 and GRK2. Pretreatment with ISO or IL-13 decreased ISO-induced cAMP generation compared to ISO for 10 min alone, paralleled by increases in β2AR and GRK2 phosphorylation. GRK2 associated with PEBP1 after 10 min of ISO in association with low pGRK2 levels. In contrast, in the presence of IL-13+ISO (10 min), binding of GRK2 to PEBP1 decreased, while 15LO1 binding and pGRK2 increased. 15LO1 knockdown restored ISO-induced cAMP generation. These findings were recapitulated in freshly brushed HAEC from asthmatic cells and tissue. IL-13 treatment of HAECs leads to β2AR desensitization which involves 15LO1/PEBP1 interactions to free GRK2 and allow it to phosphorylate (and desensitize) β2ARs, suggesting beneficial effects of β2 agonists could be blunted in Type-2 associated asthma.
登录
查看更多内容
DOI：
10.1164/rccm.200406-710st
发表时间：
2005-04-15
影响因子：
24.7
作者：
American Thoracic Society;European Respiratory Society
通讯作者：
European Respiratory Society
影响因子：
6.1
作者：
Heijink, IH;van den Berge, M;Kauffman, HF
通讯作者：
Kauffman, HF
影响因子：
64.8
作者：
Lorenz, K;Lohse, MJ;Quitterer, U
通讯作者：
Quitterer, U
影响因子：
6
作者：
Chu, HW;Balzar, S;Wenzel, SE
通讯作者：
Wenzel, SE
影响因子：
158.5
作者：
Corren, Jonathan;Lemanske, Robert F., Jr.;Matthews, John G.
通讯作者：
Matthews, John G.