Recombinant tissue plasminogen activator enhances microparticle release from mouse brain-derived endothelial cells through plasmin

Autor: Jean-Marie Renard, Kahina Khacef, Anne-Clémence Vion, Marie Garraud, Claire Leconte, Catherine Marchand-Leroux, Isabelle Margaill, Virginie Beray-Berthat, Chantal M. Boulanger, Min Yin
Přispěvatelé: Paris-Centre de Recherche Cardiovasculaire (PARCC (UMR_S 970/ U970)), Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP)
Rok vydání: 2015
Předmět:
0301 basic medicine
MAPK/ERK pathway
Time Factors
Endothelium
Plasmin
Cell Survival
[SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology
[SDV.BC]Life Sciences [q-bio]/Cellular Biology
Biology
p38 Mitogen-Activated Protein Kinases
Cell Line
03 medical and health sciences
chemistry.chemical_compound
Mice
0302 clinical medicine
Fibrinolytic Agents
Annexin
Cell-Derived Microparticles
medicine
Animals
Fibrinolysin
Endothelial dysfunction
ComputingMilieux_MISCELLANEOUS
ICAM-1
Dose-Response Relationship
Drug

Brain
Endothelial Cells
Plasminogen
medicine.disease
Molecular biology
Recombinant Proteins
Cell biology
Vascular endothelial growth factor
Endothelial stem cell
030104 developmental biology
medicine.anatomical_structure
Neurology
chemistry
Tissue Plasminogen Activator
Neurology (clinical)
Poly(ADP-ribose) Polymerases
030217 neurology & neurosurgery
medicine.drug
Signal Transduction
Zdroj: Journal of the Neurological Sciences
Journal of the Neurological Sciences, Elsevier, 2016, 370, pp.187-195. ⟨10.1016/j.jns.2016.09.026⟩
ISSN: 1878-5883
0022-510X
DOI: 10.1016/j.jns.2016.09.026⟩
Popis: Thrombolysis with recombinant tissue plasminogen activator (rt-PA) is currently the only approved pharmacological strategy for acute ischemic stroke. However, rt-PA exhibits vascular toxicity mainly due to endothelial damage. To investigate the mechanisms underlying rt-PA-induced endothelial alterations, we assessed the role of rt-PA in the generation of endothelial microparticles (EMPs), emerging biological markers and effectors of endothelial dysfunction. The mouse brain-derived endothelial cell line bEnd.3 was used. Cells were treated with rt-PA at 20, 40 or 80μg/ml for 15 or 24h, and EMPs were quantified in the culture media using Annexin-V staining coupled with flow cytometry. Rt-PA enhanced EMP release from bEnd.3 cells with a maximal increase at the 40μg/ml dose for 24h (+78% compared to controls). Using tranexamic acid and aprotinin we demonstrated that plasmin is responsible for rt-PA-induced EMP release. The p38 MAPK inhibitor SB203580 and the poly(ADP-ribose)polymerase (PARP) inhibitor PJ34 also reduced rt-PA-induced EMP production, suggesting that p38 MAPK and PARP are downstream intracellular effectors of rt-PA/plasmin. Rt-PA also altered through plasmin the morphology and the confluence of bEnd.3 cells. By contrast, these changes did not implicate p38 MAPK and PARP. This study demonstrates that rt-PA induces the production of microparticles by cerebral endothelial cells, through plasmin, p38 MAPK and PARP pathways. Determining the phenotype of these EMPs to clarify their role on the endothelium in ischemic conditions could thus be of particular interest.
Databáze: OpenAIRE