title: Correlation of in vitro and in vivo kinetics of nitric oxide donors in ocular tissues.
abstract: In the eye, nitric oxide (NO) is involved in the regulation of intraocular pressure (IOP) and ocular blood flow. The main purpose of this study was to measure the kinetics of NO release from NO donors in ocular cells and tissues using in vivo and in vitro models and demonstrate the link between the kinetics of NO release with the functional effect, IOP. Nitric oxide release was measured in human ocular cells using a fluorescent dye, diaminofluorescein (DAF), following treatment with short-acting sodium nitroprusside (SNP) and longer-acting S-nitroso-N-acetylpenicillamine (SNAP) NO donors. Both SNP and SNAP were also administered topically to rabbits; IOP was measured and levels of NO and cGMP were assessed as biomarkers over a time course in the aqueous humor (AH) and iris/ciliary body (ICB). Time- and concentration-dependent increases in NO level were produced by SNP and SNAP in human ocular cells. Both NO and cGMP levels appeared to be elevated following treatment with the aforementioned NO donors in rabbit ocular tissues. Transient IOP lowering was accompanied with these biochemical estimations in rabbits, with time of maximal effect being shifted to the right for longer-acting SNAP as compared with short-acting SNP. In vitro and in vivo NO/cGMP assay results displayed a correlation between short- and longer-acting NO donors, discriminating their respective temporal actions in the eye. Due to their translatability, the in vitro DAF assay and in vivo NO fluorometric assay can therefore be potentially useful in screening novel NO donors with different temporal/kinetic profiles.
authors.affiliation: Department of Ocular Biology, Pfizer Global Research and Development, La Jolla Laboratories, San Diego, California, USA.
authors.forename: Samantha
authors.initials: S
authors.lastname: Carreiro
authors.forename: Scott
authors.initials: S
authors.lastname: Anderson
authors.forename: Hovhannes J
authors.initials: HJ
authors.lastname: Gukasyan
authors.forename: Achim
authors.initials: A
authors.lastname: Krauss
authors.forename: Ganesh
authors.initials: G
authors.lastname: Prasanna
mesh_terms: D000287:Administration, Topical; D000818:Animals; D019842:Ciliary Arteries / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007429:Intraocular Pressure / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000008:administration & dosage / Q000493:pharmacokinetics*; D009599:Nitroprusside / Q000008:administration & dosage / Q000493:pharmacokinetics*; D011817:Rabbits; D026423:S-Nitroso-N-Acetylpenicillamine / Q000008:administration & dosage / Q000493:pharmacokinetics*; D014665:Vasodilator Agents / Q000008:administration & dosage / Q000493:pharmacokinetics*
chemical_list: D020030:Nitric Oxide Donors; D014665:Vasodilator Agents; D009599:Nitroprusside; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine; D006152:Cyclic GMP