Effects of activating and deactivating cytokines on the functionally linked tetrahydrobiopterin. No pathways in vascular smooth muscle cells

The functional relationship of nitric oxide (NO) production and synthesis of tetrahydrobiopterin (BH4), the requisite cofactor for NO synthase, was investigated in rat aortic smooth muscles cells (SMC). Inflammatory cytokines induced BH4 and NO synthesis in different ratios, IL-1 beta induced mainly NO synthesis with concomitant but limiting amounts of BH4 for maximal NO production. TNF alpha did not induce NO synthesis but induced BH4 synthesis. IFN gamma was ineffective on both the induction of NO and BH4 synthesis. TGF beta downregulated NO production but did not affect BH4 biosynthesis. IL-4 and IL-10 had no effect on both BH4 and NO synthesis. Activating cytokines strongly synergized in induction of NO production, whereas endogenous BH4 production became insufficient for maximal NO synthesis. Exogenous cofactor in the form of sepiapterin or authentic BH4, but not the natural isomer 7-BH4, enhanced NO production twofold. Inhibition of BH4 synthesis with dicumarol abolished NO production that could be restored in the presence of BH4.