Tetrahydrobiopterin levels regulate endothelial cell proliferation

Vascular abnormalities, including altered angiogenesis, are major factors contributing to the morbidity and mortality of diabetes. We hypothesized that impaired angiogenesis in diabetes results from decreased tetrahydrobiopterin (BH4)-dependent synthesis of nitric oxide (NO) by endothelial cells (EC). To test this hypothesis, we utilized EC from spontaneously diabetic BB (BBd) and nondiabetes-prone BB (BBn) rats to investigate the link between BH4 and EC proliferation. There were significant decreases in the proliferation rate and expression of proliferating cell nuclear antigen in BBd versus BBn EC, with no evidence of apoptosis in either group. Sepiapterin (a precursor of BH4 via the salvage pathway) increased BH4 synthesis and enhanced proliferation of BBd EC. The stimulating effect of sepiapterin on EC proliferation was attenuated by NG-monomethyl-L-arginine, a NO synthase inhibitor. Reducing BH4 concentrations in BBn EC caused a decrease in proliferation, which was attenuated by a long-acting NO donor. Our results suggest that BH4 levels regulate proliferation of normal EC and that a BH4 deficiency impairs NO-dependent proliferation of BBd EC.