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 |
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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 |
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