Roflumilast Inhibits Lipopolysaccharide-Induced Tumor Necrosis Factor-α and Chemokine Production by Human Lung Parenchyma

Abstract

BackgroundRoflumilast is the first phosphodiesterase-4 (PDE4) inhibitor to have been approved for the treatment of COPD. The anti-inflammatory profile of PDE4 inhibitors has not yet been explored in human lung tissues. We investigated the effects of roflumilast and its active metabolite roflumilast-N-oxide on the lipopolysaccharide (LPS)-induced release of tumor necrosis factor-alpha (TNF-α) and chemokines by human lung parenchymal explants. We also investigated roflumilast’s interaction with the long-acting β2-agonist formoterol.MethodsExplants from 25 patients undergoing surgical lung resection were incubated with Roflumilast, Roflumilast-N-oxide and formoterol and stimulated with LPS. Levels of TNF-α, chemokines (in the culture supernatants) and cyclic adenosine monophosphate (in tissue homogenates) were determined with appropriate immunoassays.ResultsRoflumilast and Roflumilast-N-oxide concentration-dependently reduced the release of TNF-α and chemokines CCL2, CCL3, CCL4, CXCL9 and CXCL10 from LPS-stimulated human lung explants, whereas CXCL1, CXCL5 and CXCL8 release was not altered. Formoterol (10 nM) partially decreased the release of the same cytokines and significantly increased the inhibitory effect of roflumilast on the release of the cytokines.ConclusionsIn human lung parenchymal explants, roflumilast and roflumilast-N-oxide reduced the LPS-induced release of TNF-α and chemokines involved in the recruitment of monocytes and T-cells but not those involved in the recruitment of neutrophils. Addition of formoterol to roflumilast provided superior in vitro anti-inflammatory activity, which may translate into greater efficacy in COPD.

Roflumilast Increases Bacterial Load and Dissemination in a Model of Pseudomononas Aeruginosa Airway Infection

“…In vitro studies on human cells also show that roflumilast inhibits TNF-a production by LPS-stimulated human mononuclear cells and inhibits both IL-8-and LTB4-induced neutrophil chemotaxis (Suzuki et al, 2015). Similar to our findings, investigation of human lung tissue explants showed that roflumilast reduced the release of TNF-a and MCP-1/CCL2 from LPS-stimulated human lung explants, whereas levels of the neutrophil-recruiting chemokines CXCL1, CXCL5, and CXCL8 were not altered (Buenestado et al, 2013). Interestingly, roflumilast alone at a higher dose (100 mg/kg) caused a significant increase in plasma and lung tissue KC/CXCL1 levels and lung tissue neutrophils (McCluskie et al, 2006).…”

CCR10+ILC2s with ILC1‐like properties exhibit a protective function in severe allergic asthma

“…Experiments with human tissues were approved by the regional independent ethics committee (Comité de Protection des Personnes Île de France VIII, Paris, France). Lung samples were obtained from 9 patients undergoing surgical resection at the Foch Hospital (Suresnes, France) for lung carcinoma who had not received preoperative anticancer chemotherapy . Lung lysates were obtained after digestion with collagenase D (Merck, Lyon, France) and filtration through a cell strainer prior to immediate flow cytometry analysis.…”

The long‐acting β2‐adrenoceptor agonist olodaterol attenuates pulmonary inflammation

“…Human lung parenchyma was obtained from 14 patients (age: 66 ± 3 years; males/females, 7/7; non-smokers: 2; smokers and ex-smokers: 8 and 4; pack years: 44 ± 6) undergoing surgical resection for lung carcinoma and who had not received recent chemo/steroid therapy. The procedure for preparation of lung explants has been described elsewhere (Hackett et al ., 2008 ; Buenestado et al ., 2013 ). Explants (5 fragments of 3–5 mm 3 , total weight ∼50–100 mg) were maintained overnight at 4°C in RPMI supplemented with 2 mM L-glutamine, 100 μg·mL −1 streptomycin and 100 U·mL −1 penicillin.…”