Nitric oxide modulates expression of matrix metalloproteinase-9 in rat mesangial cells

Background. High-output levels of nitric oxide (NO) are produced by rat mes angial cells (MCs) in response to proinflammatory cytokines such as interle ukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) by the inducible isoform of NO synthase (iNOS). We tested modulatory effects of NO on the expression and activities of matrix metalloproteinases-9 and -2 (MM P-9 and MMP-2), respectively. Temporal and spatial expression of these MMPs and their specific inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), seems to be critical in the extensive extracellular matrix (ECM) remodeling that accompanies sclerotic processes of the mesangium. Methods and Results. Using the NO donors S-Nitroso-Nacetyl-D,L-penicillamin e (SNAP) and DETA-NONOate, we found strong inhibitory effects of NO mainly on the IL-1 beta-induced MMP-9 mRNA levels. NO on its own had only weak eff ects on the expression of MMP-9 and MMP-2. The addition of the NOS inhibito r N-G-monomethyl L-arginine (L-NMMA) dose dependently increased steady-stat e mRNA levels of cytokine-induced MMP-9, suggesting that endogenously produ ced NO exerts tonic inhibition of MMP-9 expression. MMP-9 activity in condi tioned media from MCs costimulated with IL-1 beta and NO donor contained le ss gelatinolytic activity than media of cells treated with IL-1 beta alone. Exogenously added NO did not alter gelatinolytic activity of MMP-9 in cell -free zymographs. The expression levels of TIMP-1 were affected by NO simil arly to the expression of MMP-9. Conclusion. We conclude that NO modulates cytokine-mediated expression of M MP-9 and TIMP-1 in rat MCs in culture. Our results provide evidence that NO -mediated attenuation of MMP-9 gelatinolytic activity is primarily due to a reduced expression of MMP-9 mRNA, and not the result of direct inhibition of enzymatic activity.