Matrix metalloproteinase 2 and tissue inhibitors of metalloproteinases regulate human aortic smooth muscle cell migration during in vitro aging.
As a direct correlation between aging and the risk of onset of vascular disease has been universally accepted, we prepared an in vitro aging model consisting in sequential passages of human aortic smooth muscle cells (AoSMC) in order to evaluate the cell behavior changes during aging. Because matrix metalloproteinases (MMP) are actively involved in matrix remodeling and disease outcome, in our model we found active MMP-2 only in the conditioned medium of young AoSMCs, whereas aged cells showed only the inactive zymogen form of MMP-2 (pro-MMP-2). We ascribed the pro-MMP-2 activation in young cells to an increase in membrane type 1 matrix metalloproteinase (MT1-MMP) content. Furthermore, we found that transcripts coding for tissue inhibitor of metalloproteinases (TIMPs) were up-regulated in aged cells, and this increase of TIMPs could also prevent pro-MMP-2 activation in aged cells. Moreover, we demonstrated that young AoSMCs possess higher migratory capabilities than aged cells. The young AoSMC migration can be inhibited by adding TIMP-1 and TIMP-2 to the cells reproducing aged AoSMC migratory behavior. Finally, the role of MMP-2 and TIMP-2 in AoSMC migration was confirmed silencing MMP-2 and TIMP-2 in young and aged AoSMCs, respectively; therefore, in this study we showed that these enzymes play a pivotal role in the regulation of the AoSMC migration during in vitro aging.
Vigetti
Davide
D
Dipartimento di Scienze Biomediche Sperimentali e Cliniche, Università degli Studi dell'Insubria, via J.H. Dunant 5, Varese 21100, Italy.
Moretto
Paola
P
Viola
Manuela
M
Genasetti
Anna
A
Rizzi
Manuela
M
Karousou
Evgenia
E
Pallotti
Francesco
F
De Luca
Giancarlo
G
Passi
Alberto
A
D001011:Aorta / Q000166:cytology; D002465:Cell Movement; D002478:Cells, Cultured; D016922:Cellular Senescence; D004792:Enzyme Precursors / Q000378:metabolism; D018093:Gelatinases / Q000378:metabolism; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000502:physiology*; D020782:Matrix Metalloproteinases / Q000378:metabolism; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000201:enzymology* / Q000502:physiology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000502:physiology; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000502:physiology; D019714:Tissue Inhibitor of Metalloproteinases / Q000502:physiology*
D004792:Enzyme Precursors; D019715:Tissue Inhibitor of Metalloproteinase-1; D019714:Tissue Inhibitor of Metalloproteinases; D019716:Tissue Inhibitor of Metalloproteinase-2; D018093:Gelatinases; D020782:Matrix Metalloproteinases; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; C069860:progelatinase; D020778:Matrix Metalloproteinase 2