Regulation of matrix metalloproteinase expression in human vein and microvascular endothelial cells. Effects of tumour necrosis factor α, interleukin 1 and phorbol ester

Abstract

Matrix metalloproteinases (MMPs) play a role in tissue remodelling and angiogenesis. We have investigated the expression and regulation of MMP-1 (interstitial collagenase), MMP-2 (gelatinase A), MMP-3 (stromelysin 1), MMP-7 (matrilysin), MMP-9 (gelatinase B) and their inhibitors TIMP-1 and TIMP-2 in human umbilical vein, femoral vein and microvascular endothelial cells, and compared these data with those obtained with human synovial fibroblasts. Non-stimulated vein endothelial cells expressed the mRNAs for MMP-1, MMP-2, TIMP-1 and TIMP-2. MMP-3 mRNA and protein were undetectable or only weakly expressed, but could be stimulated by the inflammatory mediator tumour necrosis factor alpha (TNF alpha). The expression of MMP-3 and MMP-1 was further enhanced by phorbol 12-myristate 13-acetate (PMA). Phorbol ester also induced TIMP-1 and MMP-9, the expression of the latter being further enhanced by TNF alpha or interleukin 1 alpha (IL-1 alpha). Similar stimulatory effects were observed in microvascular endothelial cells. Hence the inflammatory mediator TNF alpha induces/enhances the production of several matrix metalloproteinases in human endothelial cells. On the other hand, MMP-2 and TIMP-2 were not affected or were affected in a variable way by TNF alpha and/or phorbol ester, suggesting a dissimilar regulation of these proteins. The cyclic AMP-enhancing agent forskolin affected the production of MMPs in a cell-type-specific way. In human vein endothelial cells it enhanced the PMA-mediated induction of MMP-9, whereas it suppressed this induction in human microvascular endothelial cells and in synovial fibroblasts. On the other hand, forskolin suppressed the PMA-mediated induction of MMP-1 and MMP-3 in synovial fibroblasts, while it enhanced or did not affect this induction in various types of human endothelial cells. These observations may have implications for future pharmacological intervention in angiogenesis.

Dextran-shelled oxygen-loaded nanodroplets reestablish a normoxia-like pro-angiogenic phenotype and behavior in hypoxic human dermal microvascular endothelium

“…In particular, cells were found to constitutively release only the 496 latent 72 kDa form of MMP-2, whereas its 62 kDa activated form was not detected. These 497 results are in line with previous reports on endothelial cells from both micro-and macro-498 vascular vessels [Hanemaaijer et al, 1993;Ben-Yosef et al, 2002;Ben-Yosef et al, 2005; 499 Bertl et al, 2006]. Exposure of endothelial cells to prolonged hypoxia led to enhanced MMP-500 2 and diminished TIMP-2 protein levels in cell supernatants, whereas TIMP-1 production 501…”

Tissue levels of matrix metalloproteinases MMP-2 and MMP-9 are related to the overall survival of patients with gastric carcinoma

“…Gelatin-zymography Presence of active and latent forms of matrix metalloproteinases was analysed by zymography on 10% polyacrylamide gels containing 2% gelatin and overnight incubation at 37°C, as described previously (Hanemaaijer et al, 1993 the carcinomas was significantly associated with a worse survival, exemplified by a shorter median survival time and a low percentage survival of the patients (Table I). The mean levels of matrix metalloproteinases MMP-2 and MMP-9, as determined by EDTA-inhibitable gelatin-zymography (Figure 1), were significantly higher in carcinomas than in histologically confirmed tumour-free adjacent mucosa of the stomach, irrespective of MMP type or activity state (Table II).…”

Convenient fluorometric assay for matrix metalloproteinase activity and its application in biological media

“…Incubation time was extended up to 6 h for samples containing low activity to acquire reliable ARFU/time data. Conditioned media (75 IXl) from untreated human umbilical vein endothelial cells cultured under serum-free conditions for 24 h [19] were incubated with 5 ~tM TNO211 in a total volume of 150 lal. Increase in fluorescence was measured as mentioned above.…”

“…Serum-free medium from non-activated human endothelial cells have been shown to contain MMPs (predominantly gelatinases [19]). When incubated with TNO211, a linear increase in fluorescence was observed within minutes, indicating (Fig.…”