Contribution of aldehyde dehydrogenase 3A1 to disulfiram penetration through monolayers consisting of cultured human corneal epithelial cells.
We previously prepared 2-hydroxypropyl-beta-cyclodextrin (HPbetaCD) solutions containing disulfiram (DSF) and hydroxypropylmethylcellulose (HPMC, DSF solutions), and found the instillation of this DSF solutions delayed lens opacification in ICR/f rats, a recessive-type hereditary cataractous strain. In this study, we determined the corneal penetration mechanism of DSF solutions using human cornea epithelial cell monolayers based on the immortalized human cornea epithelial cell line (HCE-T) developed by Tropainen et al. [Invest. Ophthalmol. Vis. Sci., 42, 2942-2948 (2001)]. The transepithelial electrical resistance (TER) values of HCE-T cells increases from approximately 275 to 388 Omega.cm(2) by exposure to an air-liquid interface for 2 weeks. The penetration of DSF into the basolateral chamber was prevented by the increase in TER values. The DSF in solution was converted to diethyldithiocarbamate (DDC) during the penetration experiment using HCE-T cell monolayers, and a close relationship between the penetration coefficient of DDC and aldehyde dehydrogenase (ALDH) 3A1 mRNA expression (y=41.202x+18.587, R=0.9413) was observed. In addition, an anti-ALDH3A1 antibody significantly inhibited the DSF-DDC conversion. These results suggest that DSF in DSF solutions is converted to DDC via catalysis by an ALDH3A1 in the cornea, and then transited from the apical side to the basolateral side.
Nagai
Noriaki
N
School of Pharmacy, Kindai University, Osaka, Japan.
Inomata
Mitsushi
M
Ito
Yoshimasa
Y
D000444:Aldehyde Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019138:Antibodies, Blocking / Q000737:chemistry / Q000494:pharmacology; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology / Q000378:metabolism*; D004221:Disulfiram / Q000493:pharmacokinetics*; D004050:Ditiocarb / Q000378:metabolism; D004847:Epithelial Cells / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012869:Skin Absorption
D019138:Antibodies, Blocking; D012313:RNA; D004050:Ditiocarb; C480021:ALDH3A1 protein, human; D000444:Aldehyde Dehydrogenase; D004221:Disulfiram