University of Manchester, Manchester University Foundation Hospital Trust, Manchester M23 9LT, UK.
University of Manchester, Manchester University Foundation Hospital Trust, Manchester M23 9LT, UK.
Cytokine. 2019 Nov;123:154739. doi: 10.1016/j.cyto.2019.154739. Epub 2019 Jul 15.
We compared the anti-inflammatory effects of phosphodiesterase type 4 (PDE4) inhibitor roflumilast with CHF6001, a novel PDE4 inhibitor designed for inhaled administration, using human alveolar macrophages (AM) and lung tissue explants models.
AM from 13 chronic obstructive pulmonary disease (COPD) patients and 10 smoking controls and lung tissue from 7 COPD patients were stimulated with LPS following preincubation with roflumilast (0.000001-10 µM), CHF6001 (0.000001-0.1 µM), or vehicle. After 24 h, supernatants were analysed for cytokines by ELISA. The effects of both compounds on the phosphorylation and cellular localisation of cAMP response element binding protein (CREB) were assessed by immunofluorescence and Western blot analysis. Extracted RNA was used for quantitative PCR analysis of PDE4 A, B and D mRNA.
PDE4 A, B and D expression were increased in alveolar macrophages and lung tissue of COPD patients compared to controls. Roflumilast and CHF6001 significantly reduced TNF-α production in AM and lung tissue. CHF6001 was more potent than roflumilast with lower ECs of 0.02, 0.01 and 0.31 nM compared to 0.87, 0.47 and 10.8 nM in respective samples. PDE4 inhibition also inhibited secretion of the chemokines CCL2 and CCL4 from macrophages. Both compounds increased nuclear levels of phosphorylated CREB.
PDE4 inhibitors caused a robust anti-inflammatory effect on TNF-α production from COPD AM, with inhibition of selective chemokines also observed. CHF6001 caused more potent inhibition of TNF-α production from COPD AM and lung tissue compared to roflumilast.
我们比较了磷酸二酯酶 4（PDE4）抑制剂罗氟司特和新型 PDE4 抑制剂 CHF6001 的抗炎作用，这两种药物都旨在通过吸入给药，使用人肺泡巨噬细胞（AM）和肺组织外植体模型。
用 LPS 刺激来自 13 例慢性阻塞性肺疾病（COPD）患者和 10 例吸烟对照者的 AM，以及来自 7 例 COPD 患者的肺组织，然后用罗氟司特（0.000001-10µM）、CHF6001（0.000001-0.1µM）或载体预孵育。24 小时后，通过 ELISA 分析上清液中的细胞因子。通过免疫荧光和 Western blot 分析评估这两种化合物对 cAMP 反应元件结合蛋白（CREB）磷酸化和细胞内定位的影响。提取的 RNA 用于定量 PCR 分析 PDE4A、B 和 DmRNA。
与对照组相比，COPD 患者的肺泡巨噬细胞和肺组织中 PDE4A、B 和 D 的表达增加。罗氟司特和 CHF6001 显著降低 AM 和肺组织中 TNF-α的产生。与各自样本中 0.87、0.47 和 10.8nM 的 EC50 相比，CHF6001 的作用更强，EC50 分别为 0.02、0.01 和 0.31nM。PDE4 抑制还抑制了巨噬细胞中趋化因子 CCL2 和 CCL4 的分泌。这两种化合物均增加了磷酸化 CREB 的核水平。
PDE4 抑制剂对 COPD AM 中 TNF-α的产生产生了强大的抗炎作用，同时也观察到对选择性趋化因子的抑制作用。与罗氟司特相比，CHF6001 更能抑制 COPD AM 和肺组织中 TNF-α的产生。
