Pro-Inflammatory Cytokines Induce Expression of Matrix-Metabolizing Enzymes in Human Cervical Smooth Muscle Cells

Abstract

The process of cervical ripening has been likened to an inflammatory reaction associated with the catabolism of cervical extracellular matrix by enzymes released from infiltrating leukocytes. We hypothesized that smooth muscle cells in the cervix also participate in this process and that pro-inflammatory cytokines act on cervical smooth muscle cells (CSMC) to provoke the expression of matrix-degrading enzymes. We treated primary cultures of human CSMC with tumor necrosis factor-␣ (TNF-␣) and examined expression of the elastinolytic enzyme , cathepsin S, the collagen metabolizing matrix metalloproteinases (MMP)-1 , -3 , -9 , and the tissue inhibitor of metalloproteinase (TIMP)-1 and -2. A time course analysis revealed that 10 ng/ml of TNF-␣ induced cathepsin S, MMP-1 , -3 , and -9 mRNA expression with the maximal response observed after 24 -48 hours. TNF-␣ induced cathepsin S , MMP-1 , -3 , and -9 mRNA expression in a dose-dependent manner: the maximal effect was observed at a concentration of 10 ng/ml , with appreciable increases observed at concentrations of 0.1 to 1.0 ng/ml. In contrast , TIMP-1 and -2 mRNAs were not significantly increased by TNF-␣ treatment. Interleukin-1␤ produced a pattern of gene expression in the CSMC similar to that observed following TNF-␣ treatment. Western blot analysis and zymography confirmed the induction of proMMP-1 , -3 , and -9 in response to TNF-␣, but MMP-2 immunoreactivity and zymographic activity were unaffected. TNF-␣ increased secretion of procathepsin S , but did not affect TIMP-1 and reduced TIMP-2 production. We conclude that CSMC are targets of pro-inflammatory cytokines, which induce a repertoire of enzymes capable of degrading the cervical extracellular matrix. The induction of these enzymes may facilitate the normal ripening of the cervix at term and participate in the The human cervix is composed primarily of connective tissue consisting mainly of fibrillar collagens, elastin, and glycosaminoglycans.

Endocrine immune interactions in human parturition

“…These enzymes include serine proteases and matrix metalloproteinases ((MMP)-8 and MMP-9)) (Kelly, 2002) which then participate in degradation of the collagen and glycosaminoglycan components of the extracellular matrix thereby leading to cervical ripening (Hertelendy and Zakar, 2004). Production of MMP-1, MMP-3 and MMP-9 from resident fibroblasts (Ito et al, 1991) and smooth muscle cells (Watari et al, 1999) may also be increased by IL-1β in the cervix whilst downregulating the expression of tissue inhibitor of metalloproteinase (TIMP)-2, an endogenous inhibitor of MMP-2 (Watari et al, 1999), with these events being further amplified by NF-κB production. NF-κB itself may be A c c e p t e d M a n u s c r i p t activated via toll-like receptor (TLR)-4 by degraded extracellular matrix proteins such as fragments of fetal fibronectin, degraded during cervical remodelling, thus reinforcing the ongoing sterile inflammation of labour (Challis et al, 2009).…”

Porcine Aortic Endothelial Cell Genes Responsive to Selected Inflammatory Stimulators

“…It plays a major role in the degradation of the invariant chain associated with the major histocompatiblity complex and thus affects antigen presentation [2]. Its overexpression by TNF- in human cervical smooth muscle cells was previously reported [33]. Bcl-2-like 14 (BCL2L14) was induced more than three-fold by TNF-.…”

Maternal Inflammatory Response in Severe Preeclamptic and Preeclamptic Pregnancies

“…Increases in pro-inflammatory cytokines (e.g. TNF-α, IL-1β) are also required for similar processes, such as by stimulating production of matrix metalloproteinase (MMP)-1, MMP-3, MMP-9 which need for cervical ripening [26,27]. In support of preeclampsia involving an exacerbation of the physiological increase of this inflammatory responsiveness, which may be due to the comparatively small numbers of patients involved.…”