Nitric oxide attenuates matrix metalloproteinase-9 production by endothelial cells independent of cGMP- or NF[kappa]B-mediated mechanisms

• Published in: Molecular and cellular biochemistry Vol. 378; no. 1-2; p. 127
• Main Authors: Meschiari, Cesar A, Izidoro-toledo, Tatiane, Gerlach, Raquel F, Tanus-santos, Jose E
• Format: Journal Article
• Language: English
• Published: New York Springer Nature B.V 01.06.2013
• Subjects: Cardiovascular disease; Cardiovascular diseases; Cellular biology; Enzymes; Nitric oxide
• ISSN: 0300-8177; 1573-4919
• DOI: 10.1007/s11010-013-1602-1

Cardiovascular diseases involve critical mechanisms including impaired nitric oxide (NO) levels and abnormal matrix metalloproteinase (MMP) activity. While NO downregulates MMP expression in some cell types, no previous study has examined whether NO downregulates MMP levels in endothelial cells. We hypothesized that NO donors could attenuate MMP-9 production by human umbilical vein endothelial cells (HUVECs) as a result of less NF[kappa]B activation or cyclic GMP (cGMP)-mediated mechanisms. We studied the effects of DetaNONOate (10â[euro]"400Â [mu]M) or SNAP (50â[euro]"400Â [mu]M) on phorbol 12-myristate 13-acetate (PMA; 10Â nM)-induced increases in MMP-9 activity (by gel zymography) or concentrations (by ELISA) as well as on a tissue inhibitor of MMPsâ[euro](TM) (TIMP)-1 concentrations (by ELISA) in the conditioned medium of HUVECs incubated for 24Â h with these drugs. We also examined whether the irreversible inhibitor of soluble guanylyl cyclase ODQ modified the effects of SNAP or whether 8-bromo-cGMP (a cell-permeable analog of cGMP) influenced PMA-induced effects on MMP-9 expression. Total and phospho-NF[kappa]B p65 concentrations were measured in HUVEC lysates to assess NF[kappa]B activation. Both NO donors attenuated PMA-induced increases in MMP-9 activity and concentrations without significantly affecting TIMP-1 concentrations. This effect was not modified by ODQ, and 8-bromo-cGMP did not affect MMP-9 concentrations. While PMA increased phospho-NF[kappa]B p65 concentrations, SNAP had no influence on this effect. In conclusion, this study shows that NO donors may attenuate imbalanced MMP expression and activity in endothelial cells independent of cGMP- or NF[kappa]B-mediated mechanisms. Our results may offer an important pharmacological strategy to approach cardiovascular diseases.[PUBLICATION ABSTRACT]