Journal: British Journal of Pharmacology

Article Title: Olodaterol shows anti‐fibrotic efficacy in in vitro and in vivo models of pulmonary fibrosis

doi: 10.1111/bph.13982

Figure Lengend Snippet: Olodaterol attenuates bleomycin‐induced lung fibrosis in mice. C57BL/6 mice received an intratracheal instillation of NaCl (Ctr, n = 18) or 0.5 mg·kg−1 bleomycin (Blm, n = 24). Olodaterol was administered each day by inhalation (1 mg·mL−1) either in a preventive regimen from day 1 to 20 (Prev, n = 12) or in a therapeutic mode from day 7 to 20 (Ther, n = 24). Analyses were performed at day 21. Body weight is expressed as % of starting weight (A). Cell count is expressed as total cell count per lung; WBC, white blood cells (B). Lung density was measured by μCT analysis (C). Ashcroft scoring was performed according to Ashcroft et al. (1988) (D). FVC was measured with flexiVent (E). Lungs were weighed before lavage. Relative lung weights are expressed as % of body weight (F). Total protein concentration in BALF was determined with a BCA assay (G). Col1A1 gene expression in lung was determined via qRT‐PCR (H). Expression of β2‐adrenoceptor (β2‐AR) protein expression was determined by elisa (I). Representative μCT pictures of mouse lungs (J). Representative images from mouse lungs (formalin‐fixed, paraffin embedded); Masson trichrome staining; 2.5× (K). Data shown are means ± SEM. *P < 0.05, significantly different as indicated. Bleomycin‐treated groups were compared with untreated groups, with unpaired Student's t‐test. Olodaterol‐treated groups were compared with Bleomycin‐treated control animals, by one‐way ANOVA followed by Fisher's LSD test for parametric data.

Article Snippet: Cells were stimulated with TGF‐β (4 ng·mL −1 ) for 48 h. Supernatants were used to determine fibrotic cytokines using appropriate elisa s. Quantikine human ET‐1 elisa was from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Cell Counting, Protein Concentration, BIA-KA, Gene Expression, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Formalin-fixed Paraffin-Embedded, Staining, Control

Journal: British Journal of Pharmacology

Article Title: Olodaterol shows anti‐fibrotic efficacy in in vitro and in vivo models of pulmonary fibrosis

doi: 10.1111/bph.13982

Figure Lengend Snippet: Olodaterol attenuates TGF‐β‐stimulated protein expression of primary HLF. Fibroblasts from control donors (HLF) and patients with IPF (IPF‐LF) were pre‐incubated with different concentrations of olodaterol and subsequently stimulated with TGF‐β (4 ng·mL−1) for 48 h in the presence of the compound. α‐SMA protein expression was measured in cell lysates by an MSD Western replacement assay (A). Pro‐collagen I C‐peptide (B), fibronectin (C) and ET‐1 (D) expression was measured in supernatants by elisa. Basal levels of 16 ng·mL−1, 1.5 μg·mL−1 and 0.2 pg·mL−1 increased to 40 ng·mL−1, 2.5 μg·mL−1 and 5 pg·mL−1 respectively. Effect of olodaterol on ET‐1 protein expression in HLF and IPF‐LF in the presence of ICI‐118,551 (30 nM) (D). Data are expressed as normalized protein expression (100% is expression with TGF‐β stimulation). Data shown are means ± SEM of n = 5 different donors for HLF and n = 5 different donors for IPF cells. Horizontal dotted line is 50% inhibition of the TGF‐β‐induced effect. Representative image of TGF‐β‐induced collagen I assembly and inhibition by 10 nM olodaterol in HLF in a ‘scar‐in‐a‐jar’ assay (E). Unstimulated cells (Ctr) were compared to TGF‐β‐stimulated cells (TGFβ) and TGFβ‐stimulated and olodaterol‐treated cells (10 nM Olodaterol).

Article Snippet: Cells were stimulated with TGF‐β (4 ng·mL −1 ) for 48 h. Supernatants were used to determine fibrotic cytokines using appropriate elisa s. Quantikine human ET‐1 elisa was from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Expressing, Control, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: International Journal of Molecular Sciences

Article Title: Serum Interleukin-18, Fetuin-A, Soluble Intercellular Adhesion Molecule-1, and Endothelin-1 in Ankylosing Spondylitis, Psoriatic Arthritis, and SAPHO Syndrome

doi: 10.3390/ijms17081255

Figure Lengend Snippet: A logistic regression model of the odds ratio of the increased serum levels of interleukin-18, decreased serum levels of fetuin-A, increased serum levels of soluble intercellular adhesion molecule-1, and decreased serum levels of endothelin-1 in the spondyloarthritis group of patients compared to controls.

Article Snippet: Serum was stored at −80 °C until analysis for IL-18, fetuin-A, sICAM-1, ET-1, IL-6, IL-23, VEGF, and EGF using a sensitive sandwich ELISA method: Human IL-18 Quantitative ELISA kit (MBL, Nagoya, Japan), Human Fetuin-A Immunoassay Quantikine ® ELISA kit, Human sICAM-1 Immunoassay Quantikine ® ELISA kit, Human ET-1 Immunoassay Quantikine ® ELISA kit, Human IL-6 Immunoassay Quantikine ® ELISA kit, Human IL-23 Immunoassay Quantikine ® ELISA kit, Human VEGF Immunoassay Quantikine ® ELISA kit, and Human EGF Immunoassay Quantikine ® ELISA kit (R&D Systems, Minneapolis, MN, USA).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Serum Interleukin-18, Fetuin-A, Soluble Intercellular Adhesion Molecule-1, and Endothelin-1 in Ankylosing Spondylitis, Psoriatic Arthritis, and SAPHO Syndrome

doi: 10.3390/ijms17081255

Figure Lengend Snippet: Serum concentrations of endothelin-1 (ET-1) in patients with spondyloarthritis (SpA), ankylosing spondylitis (AS), psoriatic arthritis (PsA), SAPHO syndrome (SAPHO), and controls.

Article Snippet: Serum was stored at −80 °C until analysis for IL-18, fetuin-A, sICAM-1, ET-1, IL-6, IL-23, VEGF, and EGF using a sensitive sandwich ELISA method: Human IL-18 Quantitative ELISA kit (MBL, Nagoya, Japan), Human Fetuin-A Immunoassay Quantikine ® ELISA kit, Human sICAM-1 Immunoassay Quantikine ® ELISA kit, Human ET-1 Immunoassay Quantikine ® ELISA kit, Human IL-6 Immunoassay Quantikine ® ELISA kit, Human IL-23 Immunoassay Quantikine ® ELISA kit, Human VEGF Immunoassay Quantikine ® ELISA kit, and Human EGF Immunoassay Quantikine ® ELISA kit (R&D Systems, Minneapolis, MN, USA).

Techniques: