Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.
Department of Pediatrics, Chung Shan Medical University Hospital, Institute of Allergy, Immunology, and Rheumatology, Taichung, Taiwan.
Environ Toxicol. 2019 Apr;34(4):476-485. doi: 10.1002/tox.22701. Epub 2019 Jan 8.
Steroid-insensitive asthma-related airway inflammation is associated with the expression of epidermal growth factor receptor (EGFR) tyrosine kinase in asthmatic bronchial epithelium. Proinflammatory cytokines IL-6 and IL-8 are related to steroid-insensitive asthma. It is currently unknown how EGFR-tyrosine kinase inhibitors (EGFR-TKIs) affects house dust mite (HDM)-induced asthma in terms of inflammatory cytokines related to steroid-resistant asthma and further signaling pathway. Cytokine expressions and EGFR signaling pathway were performed by ELISA, reverse transcriptase PCR, real-time PCR, and Western blot in cell-line models. AMP-activated protein kinase (AMPK) pathway-related inhibitors were applied to confirm the association between EGFR-TKI and AMPK pathway. HDM induced IL-6 and IL-8 in a dose-dependent manner. Both Erlotinib (Tarceva) and Osimertinib (AZD-9291) reduced the levels of HDM-stimulated IL-6 and IL-8 levels in BEAS-2B cells. AZD-9291 was more effective than Erlotinib in inhibiting phospho-EGFR, and downstream phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) and phopho-signal transducer and activator of transcription 3 (p-STAT3) pathway signaling. In addition, AMPK pathway-related inhibitor, Calcium-/calmodulin-dependent protein kinase kinase β (CaMKKβ) inhibitor, down-regulated IL-8, but EGFR-TKI had no effect on AMPK pathway. Our findings highlight EGFR-TKIs, Tarceva, and AZD-9291, attenuate HDM-induced inflammatory IL-6 and IL-8 cytokines via EGFR signaling axis pathway, but not AMPK signaling pathway.
对类固醇不敏感的哮喘相关气道炎症与哮喘支气管上皮细胞中表皮生长因子受体 (EGFR) 酪氨酸激酶的表达有关。促炎细胞因子 IL-6 和 IL-8 与对类固醇不敏感的哮喘有关。目前尚不清楚 EGFR 酪氨酸激酶抑制剂 (EGFR-TKI) 如何通过与类固醇抵抗性哮喘相关的炎症细胞因子及其进一步的信号通路来影响屋尘螨 (HDM) 诱导的哮喘。通过 ELISA、逆转录 PCR、实时 PCR 和 Western blot 在细胞系模型中检测细胞因子表达和 EGFR 信号通路。应用 AMP 激活的蛋白激酶 (AMPK) 通路相关抑制剂来确认 EGFR-TKI 与 AMPK 通路之间的关联。HDM 以剂量依赖性方式诱导 IL-6 和 IL-8。厄洛替尼（特罗凯）和奥希替尼（AZD-9291）均可降低 BEAS-2B 细胞中 HDM 刺激的 IL-6 和 IL-8 水平。与厄洛替尼相比，AZD-9291 更有效地抑制磷酸化 EGFR 及其下游磷脂酰肌醇-3-激酶/蛋白激酶 B（PI3K/AKT）和磷酸信号转导子和转录激活因子 3（p-STAT3）信号通路。此外，AMPK 通路相关抑制剂钙/钙调蛋白依赖性蛋白激酶激酶β（CaMKKβ）抑制剂可下调 IL-8，但 EGFR-TKI 对 AMPK 通路无影响。我们的研究结果强调 EGFR-TKI，特罗凯和 AZD-9291 通过 EGFR 信号轴途径减轻 HDM 诱导的炎症性 IL-6 和 IL-8 细胞因子，但不通过 AMPK 信号途径。
