Cardiovascular Drugs Inhibit MMP-9 Activity from Human THP-1 Macrophages

Abstract

It is now recognized that atherosclerosis complications are related to the unstable character of the plaque rather than its volume. Vulnerable plaques often contain a large lipid core, a reduced content of smooth muscle cells, and accumulation of inflammatory cells. Colocalization of macrophages and active matrix metalloproteinases (MMPs) is likely relevant for atherosclerotic lesion disruption. Nevertheless, MMP activity and regulation by cardiovascular drugs remains poorly defined. In this study, we evaluated the effects of avasimibe, fluvastatin, and peroxisome proliferator-activated receptor (PPAR) ligands on 92-kDa gelatinase B (MMP-9) secretion by human THP-1 macrophages. THP-1 macrophages were treated with compounds for 48 h, and secreted MMP-9 protein was quantified by immunoassay. Avasimibe, fluvastatin, and PPARalpha agonists (fenofibric acid and Wy-14643) significantly reduced, in a concentration-dependent manner, MMP-9 protein (up to 67 +/- 5% for fenofibric acid). In these assays, the PPARgamma selective agonist rosiglitazone displayed a lower efficacy than other compounds. Enzymatic activity of MMP-9 was also decreased by all cardiovascular drugs tested. MMP-9 protein/activity inhibition by cardiovascular drugs was due, at least in part, to a decrease in MMP-9 mRNA. These results show that THP-1 macrophages could be an useful cellular model to investigate effects of compounds on plaque vulnerability through MMP-9 activity.

Troglitazone but not conjugated linoleic acid reduces gene expression and activity of matrix-metalloproteinases-2 and -9 in PMA-differentiated THP-1 macrophages

“…Since CLAs are PPARγ ligands and known to inhibit NF-κB-mediated expression of pro-inflammatory genes via PPARγ in various cell lines [15,16], we studied whether CLA isomers are capable of affecting gene expression and gelatinolytic activity of MMPs in macrophages which have not yet been investigated. As a model system we used the well-established PMA-differentiated THP-1 macrophage cell model, which is widely used to investigate the effects of compounds on macrophage MMP gene expression and activity [20,35,36]. In addition, several studies dealing with CLA have used the THP-1 macrophage cell model [18,26,37].…”

Gemfibrozil decreases atherosclerosis in experimental diabetes in association with a reduction in oxidative stress and inflammation

“…This is supported by studies demonstrating PPARα expression in cells of the vessel wall, including endothelial cells [3], smooth muscle cells [4] and macrophages [5]. Furthermore, in vitro studies have demonstrated that fibrates reduce a variety of mediators implicated in atherosclerotic plaque development, including proteins involved in cell recruitment [6], cell adhesion [7], cell migration [8], foam cell formation [9] and plaque stability [10]. In addition, polymorphisms in the PPARα gene have been linked to alterations in the risk of cardiovascular disease in the absence of changes in lipid levels [11].…”

Gypenoside XLIX isolated from Gynostemma pentaphyllum inhibits nuclear factor-kappaB activation via a PPAR-alpha-dependent pathway

“…All media contained L-glutamine supplemented with penicillin (100 U/ml)/ streptomycin (100 lg/ml), and 10% (v/v) heatinactivated fetal bovine serum (FBS) in a humidified atmosphere of 5% CO 2 and 95% O 2 at 37°C. To induce monocyte differentiation into macrophage, the THP-1 monocytes were treated with PMA (400 ng/ml) for 72 h, before Wy-14643, Gyp-XLIX or vehicle (0.1% DMSO) were added, and incubated for a further 48 h in culture medium for the macrophage treatment experiments [18]. The HUVEC cells were grown in M199 medium (Invitrogen Australia) supplemented with 20% FBS, 50 lg/ml endothelial cell growth supplement (Sigma, Australia), 25 mM Hepes buffer, 2 mM L-glutamine, 100 U/ml penicillin, and 100 U/ml streptomycin.…”

“…Whereas Gyp-XLIX effectively enhanced PPARa luciferase activity, it showed minimal effects on PPAR-b=d and -c1 luciferase activity, even at high concentrations (Figure 7a). To further confirm this selectivity for PPAR-a, THP-1 derived macrophage, a cell line endogenously expressing the three PPAR isoforms [18,39], was employed mRNAs and protein were extracted from the cells and examined by RT-PCR and immunoblotting, respectively. Treatment for 48 hours with vehicle (DMSO), Gyp-XLIX and the respective positive controls for PPAR-a (Wy-14643, 50 lM), PPARb=d (bezafibrate, 100 lM) and PPAR-c (ciglitazone, 10 lM), showed comparable profiles in both PPAR-a mRNA (Figure 7b) and protein (Figure 7c) expression, with minimal or no significant increase in mRNA and protein expression for PPAR-b=d and -c.…”