A novel role of MMP2 in regulating platelet NOX2 activation.

Cristina Nocella, Vittoria Cammisotto, Simona Bartimoccia, Valentina Castellani, Lorenzo Loffredo, Daniele Pastori, Pasquale Pignatelli, Valerio Sanguigni, Francesco Violi, Roberto Carnevale

Journal: Free radical biology & medicine 2021;152():355-362

PMID: 32268176

Full Text on Elsevier Science View on PubMed

Abstract

NOX2 has a key role for cellular production of reactive oxidant species (ROS) and although the mechanism of its activation is well known, little is known about its regulation. Metallo-proteinases (MMPs) regulate numerous protein activities both in physiological and pathological conditions but their interplay with NOX2 and ROS formation is still unclear. We performed experimental studies in human platelets and polymorphonuclear leukocytes (PMNs) to investigate the interplay of MMP2 with NOX2 activity. In collagen-stimulated platelets and in PMA-stimulated PMNs from healthy subjects, an immediate burst of ROS was detected at 10 min to then decline at 20 min. Coincidentally, sNOX2-dp, a split-off product of NOX2, increased and peaked at 10 min. ROS production was persistent whereas sNOX2dp is not released in cells treated with MMP2 inhibitor compared to other MMPs inhibitors. Western blot analysis showed the highest MMP2 expression on the cell membrane 10 min after stimulation. Moreover, the co-immunoprecipitation assay confirms the interaction between MMP2 and NOX2 that formed an active immuno-complex. Treating cells with NOX2ds-tat, an inhibitor of NADPH oxidase, significantly reduced ROS formation, sNOX2-dp, MMP2 expression and MMP2-NOX2-complex, which were all restored if cells were added with HO. The study provides the first evidence that MMP2 has a key role in blunting platelet NOX2 activity and eventually ROS formation.

Address: Department of Clinical Internal, Anesthesiological and Cardiovascular Sciences, Sapienza University of Rome, Italy.; Department of General Surgery and Surgical Speciality Paride Stefanini, Sapienza University of Rome, 00161, Rome, Italy.; Department of Clinical Internal, Anesthesiological and Cardiovascular Sciences, Sapienza University of Rome, Italy; Mediterranea, Cardiocentro, 80122, Napoli, Italy.; Department of Internal Medicine, University of Rome "Tor Vergata", Rome, Italy.; Mediterranea, Cardiocentro, 80122, Napoli, Italy; Department of Medical-Surgical Sciences and Biotechnologies, Sapienza University of Rome, 04100, Latina, Italy. Electronic address: [email protected].

DOI: 10.1016/j.freeradbiomed.2020.03.033

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MeSH Terms: Humans, Hydrogen Peroxide, Matrix Metalloproteinase 2, NADPH Oxidase 2, NADPH Oxidases, Platelet Activation, Reactive Oxygen Species