Background: Dysregulated expression of diverse proteases and their specific inhibitors is critical for the increase in extracellular matrix accumulation that accompanies chronic inflammatory and sclerotic processes within the renal mesangium. Within the activating cascade of several proteases, the plasminogen system plays an important role.
Methods: We tested for modulatory effects of the nitric oxide (NO) donors S-nitroso-N-acetyl-D,L-penicillamine and DETA-NONOate, and the superoxide-generating system hypoxanthine/xanthine oxidase (HXXO) on the expression and activity of tissue plasminogen activator (tPA) by ELISA and Northern blotting.
Results: Interleukin-1beta (IL-1beta)-induced tPA and plasminogen activator inhibitor (PAI)-1 mRNA and supernatant tPA antigen were significantly inhibited by both NO donors, which resulted in a net decrease in the IL-1beta-evoked tPA enzyme activity in the conditioned media. Addition of the NO-synthase inhibitor N-monomethyl l-arginine markedly increased the cytokine-triggered tPA- and PAI-1 mRNA levels, respectively. In contrast, HXXO caused a marked amplification of the IL-1beta-induced steady-state tPA mRNA level and tPA enzyme activity that was blocked by catalase. Since MnTBAP, a superoxide dismutase mimetic, had no effects on the amplification of mRNA levels, we suggest that H2O2 is the candidate reactive oxygen species (ROS) responsible for the potentiation of IL-1beta-triggered tPA and PAI-1 expression.
Conclusions: The temporal relationship between NO and ROS generation is a critical step in the modulation of tPA and PAI-1 expression in mesangial cells and may account for a dysregulation of matrix turnover during inflammatory processes in the renal mesangium.