Arginine vasopressin transport and metabolism in the pigmented rabbit conjunctiva.
The purpose of this study was to evaluate the transepithelial transport and metabolism of arginine vasopressin (AVP) in the pigmented rabbit conjunctiva, both in the absence and presence of protease inhibitors. The apparent permeability coefficient, P(app), for 3H-AVP was determined in the modified Ussing chamber, and AVP metabolites were monitored by reversed phase HPLC using a C18 column. At 50 nM donor 3H-AVP, the P(app) in the mucosal-to-serosal (ms) direction was about five times higher than that in the opposite direction. Excess (0.1 mM) AVP decreased the P(app) for labelled AVP in the mucosal-to-serosal (ms) direction by about 50%. However, intact AVP transport showed neither concentration nor direction dependence. HPLC analysis revealed two subspecies of 3H-AVP in the receiver fluid and virtually no degradation products in the donor fluid following 3 h flux experiments. 3H-AVP transported in the ms direction underwent extensive hydrolysis (73%), which was decreased by 33% with mucosal application of 2 mM camostat mesylate (an aminopeptidase inhibitor) or by 27% with 0.5 mM leupeptin (a serine protease inhibitor). By contrast, 3H-AVP transported in the serosal-to-mucosal (sm) direction resulted in only 37% hydrolysis, and mucosal application of either inhibitor did not significantly affect the P(app) for intact AVP. These data suggest that intact AVP transport in the conjunctiva may be mediated mostly by passive diffusion and enzymatic degradation of AVP may be mediated by proteolytic enzymes present on the mucosal side of the conjunctiva.
Sun
L
L
Department of Pharmaceutical Sciences, School of Pharmacy, University of Southern California, 1985 Zonal Avenue, Los Angeles, CA 90033, USA.
Basu
S K
SK
Kim
K J
KJ
Lee
V H
VH
D000818:Animals; D001127:Arginine Vasopressin / Q000008:administration & dosage / Q000378:metabolism* / Q000493:pharmacokinetics*; D001692:Biological Transport / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003228:Conjunctiva / Q000187:drug effects / Q000378:metabolism*; D004952:Esters; D016670:Gabexate / Q000031:analogs & derivatives / Q000494:pharmacology; D006146:Guanidines; D066298:In Vitro Techniques; D007976:Leupeptins / Q000494:pharmacology; D008297:Male; D011480:Protease Inhibitors / Q000494:pharmacology; D011817:Rabbits; D014316:Tritium
D004952:Esters; D006146:Guanidines; D007976:Leupeptins; D011480:Protease Inhibitors; C034532:camostat; D014316:Tritium; D001127:Arginine Vasopressin; D016670:Gabexate; C032854:leupeptin