4b). Nuclear factor (NF)-κB signalling is also involved in TNF-α-mediated MMP-9 production, but interestingly, this pathway was not affected by atorvastatin (Fig. 4c). To determine whether the
MEK/ERK signalling pathway mediates TNF-α-induced MMP-9 production by MOVAS cells, cultures were co-incubated with a MEK inhibitor, U0126 and MMP-9 message levels assayed by quantitative RT–PCR. U0126 effectively inhibited MMP-9 production in a dose-dependent manner (Fig. 4d), indicating that the signalling via the MEK/ERK pathway is necessary for TNF-α-mediated MMP-9 production by MOVAS cells. We have identified previously three key steps in the development of coronary artery damage in a disease model of KD . These pathogenic steps include T cell activation and proliferation, production
of TNF-α and TNF-α-mediated Alvelestat price MMP-9 production. In the mouse model of KD, T cell activation triggers a massive inflammatory response characterized by marked lymphocyte proliferation and cytokine production. Local inflammation and production of TNF-α at the coronary arteries stimulates the production of MMP-9 by SMC, resulting in elastin breakdown and aneurysm formation. https://www.selleckchem.com/products/PF-2341066.html All three steps in concert lead to coronary artery damage and aneurysm formation in the animal model of KD. Atorvastatin inhibited lymphocyte proliferation in response to superantigen stimulation in a dose-dependent manner. This inhibition was also observed for production of soluble mediators of inflammation including IL-2 and TNF-α. The inhibitory effect on both proliferation and cytokine production was rescued completely by mevalonic acid, confirming that the mechanism responsible for this inhibitory activity on immune activation was at HMG-CoA reductase, a similar mechanism of action in inhibiting cholesterol metabolism. Similarly, TNF-α-induced MMP-9 production was reduced in a dose-dependent
manner in response to atorvastatin. Inhibition of ERK phosphorylation appears to be the mechanism responsible for inhibition of MMP-9 production. The ability of atorvastatin to modulate these key pathogenic steps stems from its ability to inhibit the conversion of HMG-CoA to l-mevalonate. Consistent with previous findings, our data confirm that the inhibition of T cell proliferation is dependent selleck on the mevalonate pathway, as the addition of mevalonic acid to statin-treated cells rescued the inhibitory effect observed . The inhibition of the mevalonate pathway by statins leads to the loss of isoprenoid intermediates, such as geranyl pyrophosphate and farnesyl pyrophosphate. These isoprenoid intermediates act as essential lipid attachments for the post-translational modification of several small GTP-binding proteins, one of which is Ras . The Ras/Raf/Mek/Erk pathway has been demonstrated previously to be a key element involved in T cell activation, as it is involved in production of the activator protein 1 (AP1) transcription factor.