Carrier-mediated transport of NG-nitro-L-arginine, a nitric oxide synthase inhibitor, in the pigmented rabbit conjunctiva.
In this study, the transport mechanism of NG-nitro-L-arginine (L-NA), a nitric oxide synthase inhibitor that may be useful for alleviating intraocular inflammation, was characterized in the pigmented rabbit conjunctiva. L-NA, when applied to the mucosal side of the conjunctiva, led to dose-dependent increases in the short-circuit current (Isc) at 37 degrees C but not at 4 degrees C or under the Na+-free condition. Serosally added 1 mM L-NA did not elicit any change in the Isc. Mucosally added 1 mM L-NA elicited a net absorptive Na+ flux of 0.09 microEq/(cm2.hr), comparable with the Isc change. L-NA transport at 0.1 mM in the mucosal-to-serosal (ms) direction was 22 times greater than that in the serosal-to-mucosal direction. There was a good correlation between the ms flux of L-NA and the Isc changes elicited by L-NA under the same experimental conditions. L-NA transport was saturable, with a Km of 0.35 mM and a maximal flux of 290 pmol/(cm2.min). Hill analysis of L-NA flux observed at 0.1 mM L-NA in response to varying Na+ concentrations in the mucosal bathing fluid yielded a Hill coefficient of 0.98, suggesting a 1:1 coupling between Na+ and L-NA. Moreover, ms 3H-L-NA transport was inhibited by basic amino acids (L-Arg and L-Lys) and a neutral amino acid (L-Leu), but not by an acidic amino acid (L-Glu) and the D-stereoisomer of L-NA. In the case of L-Arg, inhibition was competitive with a Ki of 0.034 mM. Taken together, the above findings are consistent with the involvement of the L-Arg transport system B0,+ in the conjunctival transport of L-NA.
Hosoya
K I
KI
Department of Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, CA 90033, USA.
Horibe
Y
Y
Kim
K J
KJ
Lee
V H
VH
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D003228:Conjunctiva / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers; D004791:Enzyme Inhibitors / Q000493:pharmacokinetics* / Q000494:pharmacology; D008297:Male; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors*; D019335:Nitroarginine / Q000493:pharmacokinetics* / Q000494:pharmacology; D011817:Rabbits; D012964:Sodium / Q000378:metabolism* / Q000494:pharmacology
D004337:Drug Carriers; D004791:Enzyme Inhibitors; D019335:Nitroarginine; D012964:Sodium; D019001:Nitric Oxide Synthase