dpp-4 activity measurement  (PPD Global Central Labs)

Journal: British Journal of Pharmacology

Article Title: Glucagon‐like peptide‐1 receptor signalling reduces microvascular thrombosis, nitro‐oxidative stress and platelet activation in endotoxaemic mice

doi: 10.1111/bph.13549

Figure Lengend Snippet: GLP‐1 receptor signalling reduces iNOS‐dependent NO formation and mRNA expression in vivo and inhibits oxidative burst in monocytes. Data were obtained from wild‐type (B6), DPP‐4 knockout (DPP4‐/‐) and GLP‐1 receptor knockout (GLP‐1r‐/‐) mice. Endotoxaemia was induced with LPS and modified by treatment with linagliptin (Lina) or liraglutide (Lira). (A, B) Whole blood HbNO levels were determined by EPR as a marker for iNOS activity. NOX activity in (C) cultured and (D) isolated (from human whole blood) monocytes was determined by chemiluminescence (using L‐012) after zymosan A stimulation. (E) qRT‐PCR was used to determine mRNA expression levels of Nox2 in cultured monocytes. The data are mean ± SEM from 6–18 (A, B); cells from at least eight different cell culture wells (C, D) or at least three different individuals (E) were used. (A) *P < 0.05 versus B6 and #P < 0.05 versus B6 + LPS; (B) *P < 0.05 versus GLP‐1r−/−; (C–E) *P < 0.05 versus control and #P < 0.05 versus +LPS.

Article Snippet: Reduced DPP‐4 activity in DPP‐4 −/− mice was confirmed by DPP‐4 activity measurement ( Supporting Information Fig. S1A and provided by Boehringer Ingelheim Pharma GmbH & Co KG (Biberach an der Riss, Germany).

Techniques: Expressing, In Vivo, Knock-Out, Modification, Marker, Activity Assay, Cell Culture, Isolation, Quantitative RT-PCR, Control

Journal: British Journal of Pharmacology

Article Title: Glucagon‐like peptide‐1 receptor signalling reduces microvascular thrombosis, nitro‐oxidative stress and platelet activation in endotoxaemic mice

doi: 10.1111/bph.13549

Figure Lengend Snippet: Time course and correlation of thrombocytopenia and fluorescent bead imaging as markers for pulmonary microvascular thrombosis. GLP‐1receptor agonist improves thrombocytopenia and LDH activity in endotoxaemia. Data were obtained from wild‐type (B6), DPP‐4 knockout (DPP4‐/‐) and GLP‐1 receptor knockout (GLP‐1r‐/‐) mice. Endotoxaemia was induced with LPS and modified by treatment with linagliptin (Lina) or liraglutide (Lira). (A) Platelet count in whole blood and fluorescence signals in lungs of wild‐type mice were measured over a 12 h time course after LPS injection. The fluorescence signal was determined after injection of fluorescent beads by using fluorescence imaging. Representative images of lungs are shown below. (B) Platelet count in endotoxaemic wild‐type (B6), DPP‐4−/− and GLP‐1r−/− mice was determined 24 h after LPS injection. (C) LDH activity was measured in serum 24 h after LPS injection. The data are mean ± SEM from (A) three mice per time point, (B) 6–21, (C) 3–5 mice per group. *P < 0.05 versus B6 and #P < 0.05 versus B6 + LPS.

Article Snippet: Reduced DPP‐4 activity in DPP‐4 −/− mice was confirmed by DPP‐4 activity measurement ( Supporting Information Fig. S1A and provided by Boehringer Ingelheim Pharma GmbH & Co KG (Biberach an der Riss, Germany).

Techniques: Imaging, Activity Assay, Knock-Out, Modification, Fluorescence, Injection

Journal: British Journal of Pharmacology

Article Title: Glucagon‐like peptide‐1 receptor signalling reduces microvascular thrombosis, nitro‐oxidative stress and platelet activation in endotoxaemic mice

doi: 10.1111/bph.13549

Figure Lengend Snippet: GLP‐1 receptor signalling improves microvascular thrombosis in endotoxaemia. Data were obtained from wild‐type (B6), DPP‐4 knockout (DPP4‐/‐) and GLP‐1 receptor knockout (GLP‐1r‐/‐) mice. Endotoxaemia was induced with LPS and modified by treatment with linagliptin (Lina) or liraglutide (Lira). (A) Microvascular thrombosis was detected by fluorescence imaging using fluorescent microbeads in endotoxaemic wild‐type (B6), DPP‐4−/− and GLP1r−/− mice. Representative images of lungs are shown below. (B) Microvascular thrombosis was further detected by fluorescence microscopy of paraffin‐embedded lung sections. Nuclei are DAPI stained (blue), and pre‐injected fluorescent beads (red) were excited at their specific wavelength. (C) Fluorescent beads and fibrinogen deposit were co‐localized in lung sections by a specific antibody staining and excitation of the beads at their specific wavelength. (B, C) Representative sections selected from at least four animals per group are shown. The data are mean ± SEM from (A‐C) 4–6 mice per group. *P < 0.05 versus B6; #P < 0.05 versus B6 + LPS; §P < 0.05 versus GLP‐1r−/−.

Article Snippet: Reduced DPP‐4 activity in DPP‐4 −/− mice was confirmed by DPP‐4 activity measurement ( Supporting Information Fig. S1A and provided by Boehringer Ingelheim Pharma GmbH & Co KG (Biberach an der Riss, Germany).

Techniques: Knock-Out, Modification, Fluorescence, Imaging, Microscopy, Staining, Injection

Journal: British Journal of Pharmacology

Article Title: Glucagon‐like peptide‐1 receptor signalling reduces microvascular thrombosis, nitro‐oxidative stress and platelet activation in endotoxaemic mice

doi: 10.1111/bph.13549

Figure Lengend Snippet: GLP‐1 receptor signalling reduces murine platelet reactivity cAMP/PKA‐dependently. Data were obtained from wild‐type (B6), DPP‐4 knockout (DPP4‐/‐) and GLP‐1 receptor knockout (GLP‐1r‐/‐) mice. Endotoxaemia was induced with LPS and modified by treatment with linagliptin (Lina) or liraglutide (Lira). (A) GLP‐1 receptor protein was detected by Western blot in platelets isolated from wild‐type mouse and in (B) platelets and monocytes isolated from human whole blood. (C, D) TF‐dependent thrombin generation was measured by calibrated automated thrombography assay in PRP of mice. Representative thrombography curves are shown. (E) Thrombin/ADP‐induced platelet aggregation was measured in PRP of mice. Representative aggregation curves are shown. Arrow indicates time point of thrombin/ADP addition. (F) cAMP‐dependent phosphorylation of VASP‐Ser157 in isolated wild‐type or (G) GLP‐1r−/− mouse platelets was detected by Western blot using a specific antibody. PGI2 was used at a concentration of 0.2 ng·mL−1. The data are mean ± SEM from (A) three animals/group (B) four human individuals (C, D) 6–11, (E) 3 and (F, G) 4–6 animals per group. (B) *P < 0.05 versus monocytes; (C, D) *P < 0.05 versus B6, #P < 0.05 versus DPP‐4−/−; (E–G) *P < 0.05 versus Ctr.

Article Snippet: Reduced DPP‐4 activity in DPP‐4 −/− mice was confirmed by DPP‐4 activity measurement ( Supporting Information Fig. S1A and provided by Boehringer Ingelheim Pharma GmbH & Co KG (Biberach an der Riss, Germany).

Techniques: Knock-Out, Modification, Western Blot, Isolation, Phospho-proteomics, Concentration Assay