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The purpose of this study was to characterize the protective effect of tetrahydrobiopterin (BH 4 ), one of the cofactors of nitric oxide (NO) synthase, against NO-induced endothelial cell injury. The addition of S-nitroso-N-acetyl-D,L-penicillamine (SNAP), a NO donor, to endothelial cells induced the release of lactate dehydrogenase (LDH), a marker for cell injury. The SNAP-induced endothelial cell injury was markedly reduced by pretreatment with sepiapterin, a precursor of BH 4  synthesis. On the other hand, exogenous BH 4  had little effect on the SNAP-induced endothelial cell injury. We recently found that NO-induced endothelial cell injury involves a part of H 2 0 2  production, since the injury was blocked by the treatment with catalase. Although BH 4  released reactive oxygen species (ROS) in cell-free conditions, the increase in intracellular BH 4  by pretreatment with sepiapterin strongly reduced H 2 0 2 -induced intracellular oxidative stress. These findings suggest that the increase in intracellular BH 4  content but not extracellular BH 4 , strongly attenuates NO-induced endothelial cell injury by at least one of the mechanisms by which BH 4  reduces H 2 0 2 -induced oxidative stress. Intracellular BH 4  seems mainly to play a role as an antioxidant or as a ROS-scavenger.

Characterization of the Protective Effect of Tetrahydrobiopterin Against S-Nitroso-N -Acetyl-D ,L-Penicillamine Induced Endothelial Cell Injury