Biol Reprod
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BOR - Papers in Press, published online ahead of print October 4, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.004721
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biolreprod.102.004721v1
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Biology of Reproduction 67, 1952-1958 (2002)
DOI: 10.1095/biolreprod.102.004721 © 2002 Society for the Study of Reproduction, Inc.

Pregnancy

Production of Matrix Metalloproteinase-9 in Lipopolysaccharide-Stimulated Human Amnion Occurs Through an Autocrine and Paracrine Proinflammatory Cytokine-Dependent System1

Fabian Arechavaleta-Velascoa, Diego Ogandob, Samuel Parrya, and Felipe Vadillo-Ortega2,b

a Center for Research on Reproduction and Women's Health, University of Pennsylvania, Philadelphia, Pennsylvania 19104 b Research Direction, Instituto Nacional de Perinatologia, Mexico D.F. 11000, Mexico

The objective of this study was to determine the presence of autocrine/paracrine regulation of matrix metalloproteinase-9 (MMP-9) expression mediated by proinflammatory cytokines in human fetal membranes. Fetal membranes obtained from women who underwent cesarean delivery before labor were manually separated into amnion and chorion layers and maintained in culture. These explants were stimulated with tumor necrosis factor {alpha} (TNF{alpha}), interleukin-1ß (IL-1ß), and either lipopolysaccharide (LPS) alone or LPS with anti-TNF{alpha} or anti-IL-1ß-neutralizing antibodies. Levels of proMMP-9 in culture media were evaluated by zymography. Enzyme-linked immunosorbant assay was performed to measure the quantity of IL-1ß, TNF{alpha}, and tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) after LPS stimulation. ProMMP-9 activity was upregulated after stimulation of the amnion by LPS, TNF{alpha}, and IL-1ß. The increased activity of proMMP-9 resulting from LPS stimulation in the amnion was blocked by the addition of TNF{alpha} neutralizing antibody but not with anti-IL-1ß. No significant effect of LPS, TNF{alpha}, or IL-1ß on proMMP-9 expression was observed in the chorion; however, the chorion produced both cytokines when stimulated with LPS. In contrast, TIMP-1 levels remained unchanged in all cultures incubated in the presence of LPS. Therefore, these data indicate that proMMP-9 is produced by the amnion but not the chorion in response to LPS. Because anti-TNF{alpha}-neutralizing antibody inhibits proMMP-9 activity in the amnion, TNF{alpha} appears to upregulate proMMP-9 production by the amnion in an autocrine fashion. Meanwhile, TNF{alpha} and IL-1ß produced by the chorion may upregulate amnionic proMMP-9 production in a paracrine manner.

1 These studies were supported by the program Carnegie-Fundacion Mexicana para la Salud for Maternal/Infant Health and a grant from the Consejo Nacional de Ciencia y Tecnologia (CONACyT 26177-M).

2 Correspondence: Felipe Vadillo-Ortega, Instituto Nacional de Perinatologia, Direccion de Investigacion, Montes Urales 800, Mexico, D.F., 11000, Mexico. FAX: 5255 5520 0034; felipe.vadillo{at}uia.mx






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Copyright © 2002 by the Society for the Study of Reproduction.