Prado, Alejandro F.
Costa, Rute A. P.
[1, 3, 4, 6, 2, 5, 7]
Paes Leme, Adriana F.
Bendhack, Lusiane M.
Tanus-Santos, Jose E.
Gerlach, Raquel F.
Total Authors: 10
 Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Pharmacol, Ribeirao Preto, SP - Brazil
 Fed Univ Para, Inst Biol Sci, Lab Struct Biol, Belem, Para - Brazil
 Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Biochem & Immunol, Ribeirao Preto, SP - Brazil
 Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Biomech Med & Rehabil Locomotor Syst, Ribeirao Preto, SP - Brazil
 CNPEM, LNBio, Brazilian Biosci Natl Lab, Mass Spectrometry Lab, Campinas, SP - Brazil
 Univ Sao Paulo, Fac Pharmaceut Sci Ribeirao Preto, Dept Phys & Chem, Ribeirao Preto, SP - Brazil
 Univ Sao Paulo, Fac Dent Ribeirao Preto, Dept Morphol Physiol & Basic Pathol, Av Cafe S-N, BR-14040904 Ribeirao Preto, SP - Brazil
Total Affiliations: 7
Web of Science Citations:
Increased reactive oxygen species (ROS) formation may enhance matrix metalloproteinase (MMP)-2 activity and promote cardiovascular dysfunction. We show for the first time that MMP-2 is upstream of increased ROS formation and activates signaling mechanisms impairing redox balance. Incubation of vascular smooth muscle cells (VSMC) with recombinant MMP-2 increased ROS formation assessed with dihydroethidium (DHE) by flow cytometry. This effect was blocked by the antioxidant apocynin or by polyethylene glycol-catalase (PEG-catalase), and by MMP inhibitors (doxycycline or GM6001). Next, we showed in HEK293 cells that MMP-2 transactivates heparin-binding epidermal growth factor (HB-EGF) leading to EGF receptor (EGFR) activation and increased ROS concentrations. This effect was prevented by the EGFR kinase inhibitor Ag1478, and by phospholipase C (PLC) or protein kinase C (PKC) inhibitors (A778 or chelerythrine, respectively), confirming the involvement of EGFR pathway in MMP-2-induce responses. Next, we showed that intraluminal exposure of aortas to MMP-2 increased vascular MMP-2 levels detected by immunofluorescence and gelatinolytic activity (by in situ zimography) in association with increased ROS formation. This effect was inhibited by MMP inhibitors (phenanthroline or doxycycline) and by apocynin or PEG-catalase. MMP-2 also increased aortic contractility to phenylephrine and this effect was prevented by MMP inhibitor GM6001 and by apocynin or PEG-catalase, showing again that increased ROS formation mediates functional effects of MMP-2. These results show that MMP-2 activates the EGFR and triggers downstream signaling pathways increasing ROS formation and promoting vasoconstriction. These findings may have various implications for cardiovascular diseases. (AU)